Sample records for blood polymorphonuclear leukocytes

  1. Apoptosis and necrosis of blood and milk polymorphonuclear leukocytes

    E-print Network

    Paris-Sud XI, Université de

    milk somatic cell counts (SCC) are used as an indicator for bovine mastitis. Dur- ing mastitisShort note Apoptosis and necrosis of blood and milk polymorphonuclear leukocytes in early, polymorphonuclear leukocytes (PMN) become the predominant cell type. Shortly after parturition, the severity

  2. Comparison of circulating phagocyte oxidative activity measured by chemiluminescence in whole blood and isolated polymorphonuclear leukocytes

    Microsoft Academic Search

    Steffi Kopprasch; J. Graessler; M. Kohl; S. Bergmann; H.-E. Schröder

    1996-01-01

    The generation of reactive oxygen species (ROS) was measured by chemiluminescence (CL) in whole blood and isolated polymorphonuclear leukocytes (PMNLs) in 113 subjects. There were no differences in ROS production in males and females, smokers and non-smokers and no age dependency. A significant correlation was found between whole blood CL parameters and between parameters derived from isolated cells, but there

  3. Enzymatic Activities of Bovine Peripheral Blood Leukocytes and Milk Polymorphonuclear Neutrophils during Intramammary Inflammation Caused by Lipopolysaccharide

    Microsoft Academic Search

    C. Prin-Mathieu; Y. Le Roux; G. C. Faure; F. Laurent; M. C. Bene; F. Moussaoui

    2002-01-01

    Leukocytes are recruited from peripheral blood into milk as part of the inflammatory response to mastitis. However, excessive accumulation of inflammatory cells alters the quality of milk and the proteases produced by polymorphonuclear neutrophils (PMNs) and macrophages may lead to mammary tissue damage. To investigate PMN recruitment and the kinetics of their intracytoplasmic enzymes in inflammation, we generated mastitis in

  4. Cardiopulmonary Bypass Primes Polymorphonuclear Leukocytes

    Microsoft Academic Search

    Jess D. Schwartz; Peter Shamamian; Daniel S. Schwartz; Eugene A. Grossi; Chad E. Jacobs; Federico Steiner; Peter C. Minneci; F. Gregory Baumann; Stephen B. Colvin; Aubrey C. Galloway

    1998-01-01

    Polymorphonuclear leukocyte (PMN) superoxide (·O?2) production has been implicated in the pathogenesis of cardiopulmonary bypass (CPB)-related end organ injury. PMN “priming” has been described as an event which enhances the release of ·O?2following a second, activating insult. We hypothesized that PMN priming occurs during CBP and is temporally related to the plasma level of complement (C3a), interleukin (IL)-6, and IL-8.

  5. Effect of viral infection of Fc receptors and immunologically nonspecific receptors of blood polymorphonuclear leukocytes: an in vitro model.

    PubMed

    Niemia?towski, M

    1989-03-01

    I describe the effect on phagocytic activity of the migration of bovine blood polymorphonuclear leukocytes (PMNL) through collagen and a monolayer of PK-15 cells in vitro infected with rotaviruses (strain OSU, RFC-17, SA-11), a picornavirus (strain SVDV), bovine herpesvirus (BHV-1) and equine herpesvirus (EHV-1). The PMNL were examined before and after their migration as follows: (1) for EA rosette formation with sensitized sheep erythrocytes, (2) for phagocytic activity mediated through Fc receptors (FcRs), (3) for phagocytic activity mediated through immunologically nonspecific receptors (INsRs). Random migration of PMNL through the infected monolayer to RPMI-1640 medium containing or lacking chemotactic agents (zymosan activated serum--Act. serum) and/or containing control agents (lipopolysaccharide--LPS or formyl-L-methionyl-L-leucyl-L-phenylalanine--fMLP) decreased the percentage of rosettes forming PMNL and the phagocytosis mediated by FcRs and INsRs. The results obtained suggest that the viruses used in this study independently of their biological properties exerted a suppressive action on the phagocytic activity mediated by FcRs and INsRs. PMID:2569805

  6. Control of vascular permeability by polymorphonuclear leukocytes in inflammation

    Microsoft Academic Search

    Caroline V. Wedmore; T. J. Williams

    1981-01-01

    Polymorphonuclear leukocytes infiltrate tissues in response to an inflammatory stimulus to remove invading microorganisms and cell debris. We present evidence that these scavenging cells have another, more sophisticated role in that they are involved in the control of fluid efflux through the blood vessel wall which leads to tissue oedema.

  7. Adhesion and motility of polymorphonuclear leukocytes isolated from the blood of rats exposed to ozone: Potential biomarkers of toxicity

    SciTech Connect

    Bhalla, D.K.; Rasmussen, R.E.; Daniels, D.S. (Univ. of California, Irvine (United States))

    1993-12-01

    Ozone (O3) exposure of rats results in airway epithelial injury and an infiltration of polymorphonuclear leukocytes (PMNs) into the lungs, suggesting alteration of PMN functions. To identify the altered PMN functions and their possible effects on epithelia, rats were exposed to air or 0.8 ppm O3 for 2 hr. PMNs were isolated from the blood and incubated with an epithelial cell line derived from rat lung (ARL-14) or primary alveolar type II cell cultures. The PMNs from the O3-exposed rats exhibited stimulated motility and spontaneous redistribution of actin filaments and adhered in a greater number to the epithelial cells when compared with the PMNs from the air-exposed rats. Actin caps usually formed at the sites of contact between the PMNs and epithelial cells, suggesting a cytoskeletal role in the inflammatory-epithelial cell interaction. By scanning electron microscopy, PMNs from air-exposed rats had features of non-motile cells. In a striking contrast to this, PMNs from O3-exposed rats revealed surface modifications, which were quite prominent at the sites of PMN-epithelial cell contacts. Despite these morphological changes, the PMNs from O3-exposed rats did not alter the epithelial resistance, a measure of paracellular permeability. In contrast to this, PMNs stimulated by phorbol myristate acetate or N-formyl-methionyl-leucyl-phenylalanine not only exhibited greater adhesion to the epithelial cells, but also caused a reduction in epithelial resistance. The changes reflecting altered morphology, motility, and adhesion of PMNs from O3-exposed rats may represent important steps in the O3-induced inflammatory response that precedes barrier disruption in vivo, but they are not associated with increased epithelial permeability in an in vitro system. Besides their mechanistic relevance, the alterations of vascular PMNs may serve as important biomarkers for detecting O3 effects.

  8. Inhibitory effect of furosemide on activation of human peripheral blood polymorphonuclear leukocytes stimulated with n-formyl-methionyl-leucyl-phenylalanine.

    PubMed

    Bialasiewicz, P; Wlodarczyk, A; Dudkiewicz, B; Nowak, D

    2004-06-01

    The clinical efficacy of inhalatory furosemide (Fu) has been extensively studied in bronchial asthma patients but there are only a few studies addressing its action on cells participating in the underlying inflammatory process. Therefore, we investigated the effect of Fu on human peripheral blood polymorphonuclear leukocytes (PMNL) at concentrations that can be achieved in the bronchial lining fluid by inhalation, i.e. 10(-5), 10(-4) and 10(-3) M. The influence of Fu on the following PMNL parameters was investigated: intracellular calcium changes ([Ca2+]i) as a part of signal transduction and luminol dependent chemiluminescence (LCL) as an indirect measure of NADPH-oxidase activation upon n-formyl-methionyl-leucyl-phenylalanine (fMLP) stimulation; chemotaxis to fMLP, phagocytosis and intracellular killing of Staphylococcus aureus. Incubation with Fu resulted in a concentration dependent reduction of Ca2+ influx and Fu (10(-3) M) decreased the main Ca2+ parameters to one half of the control values and to the level obtained in calcium-free buffer. In contrast, Fu had no effect if preincubated with the cells and then removed by washing. The LCL signal was reduced by Fu (10(-3) M) from 2000 +/- 870 to 550 +/- 440 arbitrary units [aU] (p<0.05). In contrast to the [Ca2+]i measurements, a slightly diminished LCL was also observed following preincubation with Fu and washing. No effect of Fu was found on phagocytosis and intracellular killing of St. aureus. Fu diminished chemotaxis to fMLP but at 10(-3) M it also displayed weak chemoattractant properties. The differential action of Fu on human PMNL may add to the understanding of its topical and restricted efficacy in bronchial asthma. PMID:15135322

  9. Quantification of human cytomegalovirus DNA in peripheral blood polymorphonuclear leukocytes of immunocompromised patients by the polymerase chain reaction.

    PubMed

    Zipeto, D; Baldanti, F; Zella, D; Furione, M; Cavicchini, A; Milanesi, G; Gerna, G

    1993-09-01

    Human cytomegalovirus (HCMV) DNA amplification by the polymerase chain reaction (PCR) was utilized previously for successful monitoring of HCMV infections in immunocompromised patients. However, analysis of an extended series of clinical samples revealed the relatively frequent presence of PCR inhibitors. Hence, the need for availability of an internal control of the reaction allowing identification of false negative results. Similarly, an internal standard appeared necessary for quantification of viral DNA in clinical samples. For this purpose, we constructed a recombinant DNA molecule which could be amplified by the same set of primers used for HCMV DNA amplification. Coamplification of the recombinant DNA molecule and clinical samples proved to be a simple and reliable method for verifying sample competence for amplification. In addition, coamplification of serial known amounts of the same molecule, used as internal standard, and test sample, allowed quantification of viral DNA in polymorphonuclear leukocyte samples. Quantitative monitoring of HCMV infection and antiviral treatment may provide critical indications as to whether and when to initiate or discontinue antiviral treatment in immunocompromised patients with systemic HCMV infections. PMID:8227278

  10. Concentrations of corticosteroids, leukocytes, and immunoglobulins in blood and milk after administration of ACTH to lactating dairy cattle: effects on phagocytosis of Staphylococcus aureus by polymorphonuclear leukocytes.

    PubMed

    Paape, M J; Gwazdauskas, F C; Guidry, A J; Weinland, B T

    1981-12-01

    A study was conducted to determine relationships among plasma and milk corticosteroids, milk immunoglobulins, and phagocytosis of Staphylococcus aureus by polymorphonuclear leukocytes (PMN) isolated from milk of cows given injections of 0 (saline solution only), 100, and 200 IU of ACTH. Also determined were the effects of ACTH on mobilization of PMN into milk from the mammary gland irritated by infusion of 100 ml of saline solution containing 0.1% oyster glycogen. Cows (n = 10) were injected 6 times within a 48-hour period with 0, 100, or 200 IU of ACTH. Immediately before cows were given the 5th injection, 2 mammary quarters were infused with the saline-glycogen solution. Five hours after the 6th injection, milk from infused quarters was collected, and PMN were isolated, washed, and resuspended in autologous skimmed milk collected 5 hours after the 4th injection and before the udder was infused. Isolated PMN were incubated with S aureus and percentage of phagocytosis was determined. Concentrations of total corticosteroids in plasma and milk increased after cows were given injections of 100 and 200 IU of ACTH. The concentrations of IgA, IgG1, IgG2, IgM, and bovine serum albumin in milk after 4 injections of 0, 100, and 200 IU of ACTH were similar to preinjection (base line) concentrations. Injections of 100 and 200 IU of ACTH significantly increased the concentrations of total circulating leukocytes. Concentrations of leukocytes in milk tended to increased with increasing doses of ACTH, but the differences were not significant. Injection of 100 and 200 IU of ACTH reduced phagocytosis of S aureus by PMN. After 60 minutes of incubation, phagocytosis averaged 57% and 54%, respectively, for the ACTH treatments and 70% for controls (saline only). Results indicate that injections of ACTH that result in physiologic and pharmacologic plasma concentrations of corticosteroids inhibit phagocytosis. Impairment of phagocytosis appeared to be a direct effect of corticosteroid concentration o PMN and was not due to reduced concentrations of immunolobulins. These data indicate that reduced phagocytosis by PMN could be important in increased susceptibility to disease during stress in lactating dairy cows. PMID:6280519

  11. High affinity capture and concentration of quinacrine in polymorphonuclear neutrophils via vacuolar ATPase-mediated ion trapping: Comparison with other peripheral blood leukocytes and implications for the distribution of cationic drugs

    SciTech Connect

    Roy, Caroline; Gagné, Valérie; Fernandes, Maria J.G.; Marceau, François, E-mail: francois.marceau@crchul.ulaval.ca

    2013-07-15

    Many cationic drugs are concentrated in acidic cell compartments due to low retro-diffusion of the protonated molecule (ion trapping), with an ensuing vacuolar and autophagic cytopathology. In solid tissues, there is evidence that phagocytic cells, e.g., histiocytes, preferentially concentrate cationic drugs. We hypothesized that peripheral blood leukocytes could differentially take up a fluorescent model cation, quinacrine, depending on their phagocytic competence. Quinacrine transport parameters were determined in purified or total leukocyte suspensions at 37 °C. Purified polymorphonuclear leukocytes (PMNLs, essentially neutrophils) exhibited a quinacrine uptake velocity inferior to that of lymphocytes, but a consistently higher affinity (apparent K{sub M} 1.1 vs. 6.3 ?M, respectively). However, the vacuolar (V)-ATPase inhibitor bafilomycin A1 prevented quinacrine transport or initiated its release in either cell type. PMNLs capture most of the quinacrine added at low concentrations to fresh peripheral blood leukocytes compared with lymphocytes and monocytes (cytofluorometry). Accumulation of the autophagy marker LC3-II occurred rapidly and at low drug concentrations in quinacrine-treated PMNLs (significant at ? 2.5 ?M, ? 2 h). Lymphocytes contained more LAMP1 than PMNLs, suggesting that the mass of lysosomes and late endosomes is a determinant of quinacrine uptake V{sub max}. PMNLs, however, exhibited the highest capacity for pinocytosis (uptake of fluorescent dextran into endosomes). The selectivity of quinacrine distribution in peripheral blood leukocytes may be determined by the collaboration of a non-concentrating plasma membrane transport mechanism, tentatively identified as pinocytosis in PMNLs, with V-ATPase-mediated concentration. Intracellular reservoirs of cationic drugs are a potential source of toxicity (e.g., loss of lysosomal function in phagocytes). - Highlights: • Quinacrine is concentrated in acidic organelles via V-ATPase-mediated ion trapping. • Human peripheral blood leukocytes capture and concentrate quinacrine. • Polymorphonuclear leukocytes do so with higher apparent affinity. • Polymorphonuclear are also more competent than lymphocytes for pinocytosis.

  12. The effects of space flight on polymorphonuclear leukocyte response experiment MA-032

    NASA Technical Reports Server (NTRS)

    Martin, R. R.

    1976-01-01

    In a series of studies performed at intervals from 30 day before flight to 30 days after recovery, blood samples were obtained from the three astronauts of the Apollo Soyuz Test Project and from eight control subjects. To determine the effects of space flight on polymorphonuclear leukocytes, tests were performed on blood samples obtained as quickly as possible after splashdown and on the day following recovery. The astronauts' inhalation of propellant gases and the inception of corticosteroid therapy 1 day after recovery provided an additional opportunity to investigate the possible effects of these factors on leukocyte function. Data were obtained during each time period on the total leukocyte count, differential count, leukocyte adhesion, leukocyte migration and chemotaxis, phagocytosis, and histochemical staining for leukocyte acid and alkaline phosphatase. These observations present a variety of in vitro correlates to white blood cell function within the body. Taken together, they serve as a reasonable approximation of the effects of space flight on leukocyte function.

  13. Depression of monocyte and polymorphonuclear leukocyte oxidative metabolism and bactericidal capacity by influenza A virus.

    PubMed Central

    Abramson, J S; Mills, E L; Giebink, G S; Quie, P G

    1982-01-01

    Decreased host defense against bacterial disease associated with influenza infection may be related to virus-induced changes in phagocytic cell function. Influenza A virus initiates the respiratory burst in peripheral blood monocytes and polymorphonuclear leukocytes, with a peak chemiluminescent response approximately 3 min after virus is added to the cells in vitro. Electron micrographs of phagocytic cells incubated with influenza virus demonstrated virus attached to the cell membrane and within phagocytic vacuoles. After 20 min of incubation of the virus with phagocytic cells, the chemiluminescent response to opsonized zymosan or phorbol myristate acetate was decreased by 30 to 90%. Phagocytic activity of monocytes and polymorphonuclear leukocytes incubate with influenza virus was normal, but the bactericidal activity was significantly depressed. Influenza A virus therefore stimulates an oxidative burst in monocytes as well as polymorphonuclear leukocytes, leading to a subsequent depression of the oxidative metabolic response and bactericidal capacity of the phagocytic cells. Images PMID:7054126

  14. Polymorphonuclear leukocyte activation and hemostasis in patients with essential thrombocythemia and polycythemia vera

    Microsoft Academic Search

    Anna Falanga; Marina Marchetti; Virgilio Evangelista; Alfonso Vignoli; Marina Licini; Mara Balicco; Stefano Manarini; Guido Finazzi; Chiara Cerletti; Tiziano Barbui

    2000-01-01

    Thrombohemorrhagic complications are a major cause of morbidity and mortality in patients with essential thrombocythe- mia (ET) and polycythemia vera (PV). The pathogenesis of these complications is not completely clarified. Several studies have described abnormalities of red blood cells and platelets in these patients. How- ever, no studies are available on changes in the polymorphonuclear leukocytes (PMNs), which can play

  15. Leukocyte elastase and free collagenase activity in synovial effusions: Relation to numbers of polymorphonuclear leukocytes

    Microsoft Academic Search

    G. Cohen; K. Fehr; F. J. Wagenhäuser

    1983-01-01

    In 310 freshly analysed synovial fluid (SF) samples from patients with rheumatic and non-rheumatic diseases, activities of leukocyte elastase, collagenase and activator for human leukocyte latent collagenase were determined. These activities were investigated in relation to the number of polymorphonuclear leukocytes (PMN) and the activity of lactate dehydrogenase (LDH). Special care was taken to avoid artificial leakage of enzymes from

  16. Defense of the Bovine Mammary Gland by Polymorphonuclear Neutrophil Leukocytes

    Microsoft Academic Search

    Max Paape; Jalil Mehrzad; Xin Zhao; Johann Detilleux; Christian Burvenich

    2002-01-01

    The primary phagocytic cells of the bovine mammary gland, polymorphonuclear neutrophil leukocytes (PMN), and macrophages, comprise the first line of defense against invading pathogens. In the normal healthy mammary gland, macrophages predominate and act as sentinels to invading mastitis-causing pathogens. Once invaders are detected, macrophages, and possibly mammary epithelial cells, release chemoattractants that direct migration of PMN into the area.

  17. Effect of foscarnet induction treatment on quantitation of human cytomegalovirus (HCMV) DNA in peripheral blood polymorphonuclear leukocytes and aqueous humor of AIDS patients with HCMV retinitis. The Italian Foscarnet Study Group.

    PubMed

    Gerna, G; Baldanti, F; Sarasini, A; Furione, M; Percivalle, E; Revello, M G; Zipeto, D; Zella, D

    1994-01-01

    The aim of this study was to investigate peripheral blood polymorphonuclear leukocytes and, whenever possible, aqueous humor from 65 AIDS patients with ophthalmoscopically diagnosed human cytomegalovirus (HCMV) retinitis to determine (i) whether patients consistently carry viral DNA and (ii) to what extent foscarnet induction treatment decreases viral DNA levels. HCMV DNA was quantified by PCR using densitometric analysis of hybridization products obtained from external standards and a standard curve from which the number of genome equivalents of test samples, normalized by using an internal amplification control, was interpolated. Results showed that 56 of 65 patients (86.1%) were positive for HCMV DNA prior to induction treatment. Of 41 of the 56 patients (73.2%) whose blood had become DNA negative after induction, only 5 had a high viral load (> 5,000 genome equivalents per 2 x 10(5) polymorphonuclear leukocytes) prior to induction, whereas as many as 13 of the 15 (26.8%) patients remaining DNA positive after induction had a high viral load prior to induction. Finally, of the nine patients (13.8%) with DNA-negative blood prior to induction treatment, three were shifted to foscarnet from ganciclovir, while six were erroneously enrolled in the study. Pre- and postinduction aqueous humor samples were obtained from 12 patients; all of these were DNA positive prior to induction, whereas after induction, 4 became negative, 6 showed a marked decrease in viral DNA, and 2 had nearly stable low DNA levels. In conclusion, PCR is a valuable tool for etiologic diagnosis and monitoring of HCMV retinitis treatment in AIDS patients. HCMV DNA is consistently present in the blood and aqueous humor of all patients with HCMV retinitis. Foscarnet induction treatment is highly effective in suppressing or reducing DNA levels in both blood leukocytes and aqueous humor. PMID:8141577

  18. Effect of foscarnet induction treatment on quantitation of human cytomegalovirus (HCMV) DNA in peripheral blood polymorphonuclear leukocytes and aqueous humor of AIDS patients with HCMV retinitis. The Italian Foscarnet Study Group.

    PubMed Central

    Gerna, G; Baldanti, F; Sarasini, A; Furione, M; Percivalle, E; Revello, M G; Zipeto, D; Zella, D

    1994-01-01

    The aim of this study was to investigate peripheral blood polymorphonuclear leukocytes and, whenever possible, aqueous humor from 65 AIDS patients with ophthalmoscopically diagnosed human cytomegalovirus (HCMV) retinitis to determine (i) whether patients consistently carry viral DNA and (ii) to what extent foscarnet induction treatment decreases viral DNA levels. HCMV DNA was quantified by PCR using densitometric analysis of hybridization products obtained from external standards and a standard curve from which the number of genome equivalents of test samples, normalized by using an internal amplification control, was interpolated. Results showed that 56 of 65 patients (86.1%) were positive for HCMV DNA prior to induction treatment. Of 41 of the 56 patients (73.2%) whose blood had become DNA negative after induction, only 5 had a high viral load (> 5,000 genome equivalents per 2 x 10(5) polymorphonuclear leukocytes) prior to induction, whereas as many as 13 of the 15 (26.8%) patients remaining DNA positive after induction had a high viral load prior to induction. Finally, of the nine patients (13.8%) with DNA-negative blood prior to induction treatment, three were shifted to foscarnet from ganciclovir, while six were erroneously enrolled in the study. Pre- and postinduction aqueous humor samples were obtained from 12 patients; all of these were DNA positive prior to induction, whereas after induction, 4 became negative, 6 showed a marked decrease in viral DNA, and 2 had nearly stable low DNA levels. In conclusion, PCR is a valuable tool for etiologic diagnosis and monitoring of HCMV retinitis treatment in AIDS patients. HCMV DNA is consistently present in the blood and aqueous humor of all patients with HCMV retinitis. Foscarnet induction treatment is highly effective in suppressing or reducing DNA levels in both blood leukocytes and aqueous humor. Images PMID:8141577

  19. Calprotectin, an Abundant Cytosolic Protein from Human Polymorphonuclear Leukocytes, Inhibits the Growth of Borrelia burgdorferi

    Microsoft Academic Search

    Denise Lusitani; Stephen E. Malawista; Ruth R. Montgomery

    2003-01-01

    We previously showed that numerous polymorphonuclear leukocyte (PMN) granule components efficiently kill Borrelia burgdorferi, the agent of Lyme disease. In addition, motile, granule-poor cytoplasts (U-Cyt) from human blood PMN can exert anti-Borrelia activity against opsonized B. burgdorferi independently of oxidative mechanisms. Here we show that lysates of U-Cyt also possess anti-Borrelia activity, a portion of which comes from the abundant

  20. Leukocyte Elastase Release during Blood Coagulation

    PubMed Central

    Plow, Edward F.

    1982-01-01

    Immunological detection of elastase, an enzyme present within leukocyte granules, has been used as a marker for polymorphonuclear leukocyte activation. Polymorphonuclear leukocytes contained 4.6 ?g/107 cells, whereas erythrocytes, mononuclear cells, and platelets contained <1% of this level. In plasma that was separated from blood cells after 1 h at 22°C, the mean level of elastase-related antigen in seven normal donors was 25±6 ng/ml. This level was unaltered by immediate separation of the plasma from the cells, by inclusion of protease inhibitors, or by anticoagulation of the plasma with either EDTA or acidcitrate-dextrose (the level in heparinized plasma was approximately threefold higher). In serum, the level of elastase-related antigen was 288±125 ng/ml, representing an 11.5-fold increase above plasma levels. The antigen detected in serum was immunochemically indistinguishable from the leukocyte enzyme. Release of elastase was observed when isolated polymorphonuclear leukocytes were added to nonanticoagulated platelet-rich or platelet-poor plasma, recalcified plasma, or to serum. Addition of a chelating agent to serum prevented elastase release, but calcium or magnesium did not induce release in the absence of plasma. Coagulation induced by addition of thrombin to plasma also failed to induce release. In whole blood or in anticoagulated plasma reconstituted with polymorphonuclear leukocytes and then recalcified, initial release of elastase occurred concomitantly with or slightly after clotting and reached maximal levels within 20-40 min after clot formation. The data indicates that early events in coagulation or other pathways that occur in parallel with coagulation induce leukocyte release. The release of elastase, a major fibrinolytic protease of leukocytes, from the cells provides a mechanism for this enzyme or other granule proteases to participate in physiological events. Images PMID:6916769

  1. Nerve growth factor: stimulation of polymorphonuclear leukocyte chemotaxis in vitro.

    PubMed Central

    Gee, A P; Boyle, M D; Munger, K L; Lawman, M J; Young, M

    1983-01-01

    Topical application of mouse nerve growth factor (NGF) to superficial skin wounds of mice has previously been shown to accelerate the rate of wound contraction. Results of the present study reveal that NGF in the presence of plasma is also chemotactic for human polymorphonuclear leukocytes in vitro, and the concentration of NGF required for this effect is similar to that which stimulates ganglionic neurite outgrowth. This property does not arise from liberation of the C5a fragment of complement, nor does it require the known enzymic activity of NGF. (NGF inactivated with diisopropyl fluorophosphate is equally active.) We conclude that NGF can display biological effects on cells of nonneural origin and function, and this feature might play a role in the early inflammatory response to injury. PMID:6580641

  2. Chronic renal failure: leukocyte rheology and polymorphonuclear cytosolic Ca2+ concentration.

    PubMed

    Lo Presti, R; Canino, B; Vaccaro, F; Montana, M; Ventimiglia, G; Grifň, G; Caimi, G

    1999-01-01

    In a group of subjects with chronic renal failure (CRF), we evaluated the leukocyte rheological activity, expressed as the filtration rate, the polymorphonuclear (PMN) membrane fluidity and the PMN cytosolic Ca2+ content. Using the St. George's Filtrometer, we determined the filtration rate of unfractionated, mononuclear and PMN leukocytes. Using the fluorescent probe 1.4-(trimethylamino)-phenyl-4-phenyl-hexatriene (TMA-DPH), we examined the PMN membrane fluidity and, using the Fura 2-AM, the PMN cytosolic Ca2+ content. From the results obtained, it is evident that only the initial relative flow rate of unfractionated leukocytes was significantly reduced in subjects with CRF, while the filtration parameters of mononuclear and PMN cells did not distinguish normals from CRF subjects. No variation was evident for the PMN membrane fluidity, while the PMN cytosolic Ca2+ content was significantly increased in CRF subjects. In these subjects no correlation was found between leukocyte filtration parameters, PMN membrane fluidity, PMN cytosolic Ca2+ content and plasma parameters (blood urea nitrogen and serum creatinine), reflecting the degree of the CRF. In conclusion, in CRF subjects the abnormality of the leukocyte flow properties was restricted to the initial flow rate of unfractionated leukocytes, whereas the increase of PMN cytosolic Ca2+ content might be attributed to PMN activation. PMID:10621927

  3. Age-related comparison of superoxide production by canine neutrophilic polymorphonuclear leukocytes

    E-print Network

    Hanson, Thomas Dale

    1983-01-01

    AGE-RELATED COMPARISON OF SUPEROXIDE PRODUCTION BY CANINE NEUTROPHILIC POLYMORPHONUCLEAR LEUKOCYTES A Thesis by THOMAS DALE HANSON Submitted to the Graduate College of Texas A&M University in partial fulfillment of the requirements... for the degree of MASTER OF SCIENCE December 1983 Major Subject: Veterinary Microbiology AGE-RELATED COMPARISON OF SUPEROXIDE PRODUCTION BY CANINE NEUTROPHILIC POLYMORPHONUCLEAR LEUKOCYTES A Thesis by THOMAS DALE HANSON Approved as to style and content...

  4. Effects of mercury on human polymorphonuclear leukocyte function in vitro.

    PubMed Central

    Contrino, J.; Marucha, P.; Ribaudo, R.; Ference, R.; Bigazzi, P. E.; Kreutzer, D. L.

    1988-01-01

    A variety of heavy metals are recognized as environmental pollutants, and although a significant body of literature exists on the acute toxicity of these metals in various tissues, little is known about the effects of metals such as mercury on host defense. Therefore, the effect of mercuric chloride (HgCl2) on human polymorphonuclear leukocytes (PMN) function in vitro was evaluated. The acute toxicity of HgCl2 for human PMN was calculated initially using vital dye exclusion (trypan blue), and lactate dehydrogenase (LDH) release. Concentrations of HgCl2 less than or equal to 10(-6) M did not induce significant LDH release, or uptake of trypan blue. Additionally, HgCl2 at less than or equal to 10(-7) M produced no ultrastructural alterations in the PMN. The effects of HgCl2 on human PMN functions involved in host defense were evaluated next. HgCl2 consistently suppressed human PMN adherence, polarization, chemotaxis, and erythrophagocytosis at concentrations between 10(-6) and 10(-17) M. Because of the established role of oxygen metabolites in host defense, the effects of HgCl2 on human PMN chemiluminescence and H2O2 production were evaluated next. These studies demonstrated that low concentrations of HgCl2 (ie, 10(-9)-10(-15) M) significantly enhanced chemiluminescence, as well as stimulated H2O2 production by the PMN. These studies clearly demonstrate the ability of extremely low levels of HgCl2 not only to suppress various PMN functions involved in host defense, but also to stimulate oxygen metabolism. In vivo, these HgCl2 effects would not only compromise host defense but also promote tissue injury via the local production of oxygen metabolites. Images Figure 1 Figure 2 PMID:3394794

  5. [Possibility of diagnostics of the non-steroidal anti-inflammatory drugs intolerance with a change in the chemiluminescent glow of the polymorphonuclear leukocytes of the peripheral blood].

    PubMed

    Chausova, S V; Gurevich, K G; Bondareva, G P; Filatov, O Ju; Malyshev, I Y

    2014-01-01

    We investigated the intensity of barium sulfate stimulated luminol-dependent chemiluminescence (SLCL) of pre-incubated blood with various concentrations of sodium salicylate, sodium metamizol or sodium diclofenac. Blood was received from healthy donors and patients with intolerance to aspirin and/or sodium metamizol and/or sodium diclofenac. Revealed valid differences in SLCL of blood received from healthy donors and patients with intolerance to these drugs allows us to use chemiluminescence method for the diagnosis of intolerance to non-steroidal anti-inflammatory drugs. PMID:25980237

  6. Inhibition of superoxide production in human polymorphonuclear leukocytes by oral treponemal factors.

    PubMed Central

    Sela, M N; Weinberg, A; Borinsky, R; Holt, S C; Dishon, T

    1988-01-01

    The inhibition of superoxide (O2-) production by human peripheral blood polymorphonuclear leukocytes (PMNs) in the presence of oral treponemes, their cellular components, and their culture supernatants was investigated. Superoxide production was inhibited 56% by a 25-microgram/ml phenol extract of a human clinical isolate. Inhibition by culture supernatants of both the clinical isolate and a reference strain was related to the bacterial phase of growth and viability, though inhibition also persisted in the decline phase. Inhibition of superoxide production was not evident when either opsonized or nonopsonized whole spirochetes were reacted with PMNs. The suppressive activity depended, therefore, on the treponemes either being disrupted or growing and releasing the inhibitory factor into the culture medium. These results suggest that oral treponemes possess factors which interfere with the activity of PMNs and thereby alter the inflammatory process in the diseased periodontal pocket. PMID:2830192

  7. Lactoferrin in human neutrophilic polymorphonuclear leukocytes in relation to iron metabolism.

    PubMed

    de Vet, B J; ten Hoopen, C H

    1978-01-01

    Lactoferrin (LF), the iron-binding protein of external secretions and neutrophilic polymorphonuclear leukocytes (PMN), was studied in 27 patients during granulocytosis caused by acute inflammation and in disorders without granulocytosis (iron deficiency anemia, iron overload and liver diseases). During granulocytosis the LF concentration of PMN was significantly lower than in controls (p less than 0.001). This difference proved to be related to the number of PMN. A relation between the LF concentration of PMN and iron metabolism could be demonstrated: loss of iron by blood donation is accompanied by a significant decrease in the LF concentration in PMN, whereas iron therapy in patients with iron deficiency anemia is accompanied by a significant increase in the LF concentration in PMN. PMID:416654

  8. Effect of human polymorphonuclear and mononuclear leukocytes on chromosomal and plasmid DNA of Escherichia coli. Role of acid DNase

    SciTech Connect

    Rozenberg-Arska, M.; van Strijp, J.A.; Hoekstra, W.P.; Verhoef, J.

    1984-05-01

    Phagocytosis and killing by polymorphonuclear and mononuclear leukocytes are important host resistance factors against invading microorganisms. Evidence showing that killing is rapidly followed by degradation of bacterial components is limited. Therefore, we studied the fate of Escherichia coli DNA following phagocytosis of E. coli by polymorphonuclear and mononuclear leukocytes. (/sup 3/H)Thymidine-labeled, unencapsulated E. coli PC2166 and E. coli 048K1 were incubated in serum, washed, and added to leukocytes. Uptake and killing of the bacteria and degradation of DNA were measured. Although phagocytosis and killing by mononuclear leukocytes was less efficient than that by polymorphonuclear leukocytes, only mononuclear leukocytes were able to degrade E. coli PC2166 DNA. Within 2 h, 60% of the radioactivity added to mononuclear leukocytes was released into the supernate, of which 40% was acid soluble. DNA of E. coli 048K1 was not degraded. To further analyze the capacity of mononuclear leukocytes to degrade E. coli DNA, chromosomal and plasmid DNA was isolated from ingested bacteria and subjected to agarose gel-electrophoresis. Only chromosomal DNA was degraded after phagocytosis. Plasmid DNA of E. coli carrying a gene coding for ampicillin resistance remained intact for a 2-h period after ingestion, and was still able to transform recipient E. coli cells after this period. Although we observed no DNA degradation during phagocytosis by polymorphonuclear leukocytes, lysates of both polymorphonuclear and mononuclear leukocytes contained acid-DNase activity with a pH optimum of 4.9. However, the DNase activity of mononuclear leukocytes was 20 times higher than that of polymorphonuclear leukocytes. No difference was observed between DNase activity from polymorphonuclear and mononuclear leukocytes from a chronic granulomatous disease patient with DNase activity from control polymorphonuclear and mononuclear leukocytes.

  9. Modulation of polymorphonuclear leukocyte adherence by pulpotomy medicaments: effects of formocresol, glutaraldehyde, eugenol, and calcium hydroxide

    Microsoft Academic Search

    W. Kim Seow; Y. H. Thong

    1986-01-01

    Activated polymorphonuclear leukocytes (PMNs) release lysosomal enzymes and toxic oxygen-free radicals into their immediate environment. The persistent activation of PMNs by pulpotomy medicaments may contribute to the chronic inflammatory changes and root resorption seen in histologic sections. The authors examined the effects of pulpotomy medicaments commonly used in pediatric dentistry on PMN adherence, the earliest observable change in PMN behavior

  10. Pseudomonas aeruginosa tolerance to tobramycin, hydrogen peroxide and polymorphonuclear leukocytes is quorum-sensing dependent

    Microsoft Academic Search

    Thomas Bjarnsholt; Peter Řstrup Jensen; Mette Burmřlle; Morten Hentzer; Janus A. J. Haagensen; Hans Petter Hougen; Henrik Calum; Kit G. Madsen; Claus Moser; Sřren Molin; Niels Hřiby; Michael Givskov

    2005-01-01

    The opportunistic human pathogen Pseudomonas aeruginosa is the predominant micro-organism of chronic lung infections in cystic fibrosis (CF) patients. P. aeruginosa colonizes the CF lungs by forming biofilm structures in the alveoli. In the biofilm mode of growth the bacteria are highly tolerant to otherwise lethal doses of antibiotics and are protected from bactericidal activity of polymorphonuclear leukocytes (PMNs). P.

  11. The influence of polymorphonuclear leukocytes on altered pulmonary epithelial permeability during ozone exposure

    Microsoft Academic Search

    Paul G Reinhart; David J. P Bassett; Deepak K Bhalla

    1998-01-01

    Ozone (O3), a pulmonary irritant, and a major toxic component of photochemical smog, is capable of inducing pulmonary inflammation characterized by recruitment of polymorphonuclear leukocytes (PMNs) into the lung. The recruited PMNs, in turn, can release toxic mediators and produce lung injury. The mechanism of ozone-induced changes in lung permeability remains unknown. It is our hypothesis that PMNs migrating into

  12. Human Polymorphonuclear Leukocytes Inhibit Aspergillus fumigatus Conidial Growth by Lactoferrin-Mediated Iron Depletion1

    Microsoft Academic Search

    Kol A. Zarember; Janyce A. Sugui; Yun C. Chang; Kyung J. Kwon-Chung; John I. Gallin

    Aspergillus fumigatus, a common mold, rarely infects humans, except during prolonged neutropenia or in cases of chronic gran- ulomatous disease (CGD), a primary immunodeficiency caused by mutations in the NADPH oxidase that normally produces fungicidal reactive oxygen species. Filamentous hyphae of Aspergillus are killed by normal, but not CGD polymorphonuclear leukocytes (PMN); however, the few studies on PMN-mediated host defenses

  13. SEQUENTIAL DEGRANULATION OF THE TWO TYPES OF POLYMORPHONUCLEAR LEUKOCYTE GRANULES DURING PHAGOCYTOSIS OF MICROORGANISMS

    Microsoft Academic Search

    DOROTHY FORD BAINTON

    1973-01-01

    The sequential discharge of neutrophilic polymorphonuclear leukocyte (PMN) granules- azurophils and specifics-was investigated by electron microscopy and cytochemistry . Thus the enzyme content of PMN phagocytic vacuoles was determined at brief intervals after phagocytosis of bacteria, utilizing peroxidase as a marker enzyme for azurophil gran- ules, and alkaline phosphatase for specifics . At 30 s, approximately half the phagocytic vacuoles

  14. FACTORS AFFECTING THE REDISTRIBUTION OF SURFACE-BOUND CONCANAVALIN A ON HUMAN POLYMORPHONUCLEAR LEUKOCYTES

    Microsoft Academic Search

    GRAEME B. RYAN; JOAN Z. BORYSENKO; MORRIS J. KARNOVSKY

    1974-01-01

    Human neutrophil polymorphonuclear leukocytes (PMN) were studied to deter- mine the influence of cellular locomotion upon the redistribution and capping of concanavalin A (Con A) . Con A was detected by fluorescence (using Con A conjugated to fluorescein isothiocyanate (Con A-FITC)), or on shadow-cast replicas (using Busycon canaliculatum hemocyanin as a marker for Con A). After labeling with Con A

  15. Functional and metabolic studies of polymorphonuclear leukocytes in the canine homologue of congenital Pelger-Huet Anomaly

    E-print Network

    Browder, Elizabeth Jarratt

    1985-01-01

    FUNCTIONAL AND METABOLIC STUDIES OF POLYMORPHONUCLEAR LEUKOCYTES IN THE CANINE HOMOLOGUE OF CONGENITAL PELGER-HUET ANOMALY A Thesis by ELIZABETH JARRATT BROWDER Submitted to the Graduate College of Texas A8M University in partial fulfillment... of requirements for the degree of MASTER OF SCIENCE May 1985 Major Subject: Veterinary Microbiology FUNCTIONAL AND METABOLIC STUDIES OF POLYMORPHONUCLEAR LEUKOCYTES IN THE CANINE HOMOLOGUE OF CONGENITAL PELGER-HUET ANOMALY A Thesis by Elizabeth Jarratt...

  16. Hydrogen Peroxide Generation by Polymorphonuclear Leukocytes Exposed to Peritoneal Dialysis Effluent

    Microsoft Academic Search

    I. DANIELS; K. S. S. BHATIA; C. J. PORTER; M. A. LINDSAY; A. G. MORGAN; R. P. BURDEN

    1996-01-01

    In the presence of peritoneal dialysis effluent (PDE), human polymorphonuclear leukocytes (PMN) showed reduced production of hydrogen peroxide and hypochlorous acid (H2O2and HOCl, respectively) when at rest and when stimulated with both soluble (formylmethionyl-leucyl-phenylalanine and phorbol myristate acetate) and particulate (Staphylococcus epidermidis) agonists. This effect occurred in a concentration-dependent man- ner between 0 and 70% (vol\\/vol) dialysis effluent. The inhibition

  17. Oxidation of proteins in rat heart and lungs by polymorphonuclear leukocyte oxidants

    Microsoft Academic Search

    Henry Fliss

    1988-01-01

    The ability of the polymorphonuclear leukocyte (PMN) oxidants, hypochlorous acid (HOC1) and hydrogen peroxide (H2O2), to oxidize proteins in rat heart and lung tissues was investigated. Cardiac myocytes, heart tissue slices, isolated perfused hearts, and lung tissue slices, were treated with HOCI and H2O2 and the extent of methionine and cysteine oxidation was determined in the cellular proteins. Cardiac tissues

  18. Dendritic Cells Take up and Present Antigens from Viable and Apoptotic Polymorphonuclear Leukocytes

    Microsoft Academic Search

    Carlos Alfaro; Natalia Suarez; Carmen Ońate; Jose L. Perez-Gracia; Ivan Martinez-Forero; Sandra Hervas-Stubbs; Inmaculada Rodriguez; Guiomar Perez; Elixabet Bolańos; Asis Palazon; Miguel Fernandez de Sanmamed; Aizea Morales-Kastresana; Alvaro Gonzalez; Ignacio Melero

    2011-01-01

    Dendritic cells (DC) are endowed with the ability to cross-present antigens from other cell types to cognate T cells. DC are poised to meet polymorphonuclear leukocytes (PMNs) as a result of being co-attracted by interleukin-8 (IL-8), for instance as produced by tumor cells or infected tissue. Human monocyte-derived and mouse bone marrow-derived DC can readily internalize viable or UV-irradiated PMNs.

  19. An evaluation of the role of leukocytes in the pathogenesis of experimentally induced corneal vascularization. III. Studies related to the vasoproliferative capability of polymorphonuclear leukocytes and lymphocytes.

    PubMed Central

    Fromer, C. H.; Klintworth, G. K.

    1976-01-01

    Studies in the past have suggested that leukocytes are a prerequisite to corneal vascularization. To test this hypothesis further, experiments were conducted to determine whether the intracorneal instillation of polymorphonuclear leukocytes, lymphocytes, or components of leukocytes would induce a corneal vascular ingrowth. These cells or cellular fractions were injected intracorneally into Fisher albino rats whose circulating leukocytes had been depleted by total body x-irradiation. Polymorphonuclear leukocytes isolated from glycogen-induced peritoneal exudates caused a corneal vascular invasion, but lymphocytes obtained from thymus, spleen, and lymph nodes failed to do so. To learn whether an extractable factor could be isolated from polymorphonuclear leukocytes these cells were suspended in isotonic saline, ultrasonified and then centrifuged at 101,952g for 1 hour. Aliquots of the resulting sediment and supernatant were injected intracorneally into rats with radiation-induced leukopenia. The nonsedimentable supernatant caused corneal vascularization, but the sediment did not provoke the phenomenon. These studies not only provide further support for the hypothesis that leukocytes initiate corneal vascularization, possibly by the release of one or more heat labile chemical mediators, but directly implicate the polymorphonuclear leukocyte in this process. Images Figures 1-3 Figure 4 Figure 5 Figure 6 PMID:1247083

  20. Production of chemotactic activity for polymorphonuclear leukocytes by cultured rat hepatocytes exposed to ethanol.

    PubMed Central

    Perez, H D; Roll, F J; Bissell, D M; Shak, S; Goldstein, I M

    1984-01-01

    Acute alcoholic hepatitis is characterized by infiltration of the liver parenchyma with polymorphonuclear leukocytes. As a possible explanation for this phenomenon, we have found that ethanol stimulates cultured rat hepatocytes to generate potent chemotactic activity. Hepatocytes (greater than 99% pure), isolated from the livers of Sprague-Dawley rats, responded to incubation with ethanol (2.0-10 mM) by releasing chemotactic activity for human polymorphonuclear leukocytes into culture supernatants in a time- and concentration-dependent fashion. Chemotactic activity was maximal after incubation of hepatocytes with 10 mM ethanol for 6 h. It was undetectable in the absence of ethanol and was reduced in the presence of either the alcohol dehydrogenase inhibitor, 4-methylpyrazole, or the acetaldehyde dehydrogenase inhibitor, cyanamide. Ethanol failed to stimulate generation of chemotactic activity by either rat dermal fibroblasts, hepatic sinusoidal endothelial cells, or Kupffer cells. The chemotactic activity generated by ethanol-treated rat hepatocytes was recovered from culture supernatants in the lipid phase after extraction with chloroform/methanol. Thin-layer chromatography and high performance liquid chromatography of chloroform/methanol extracts demonstrated that the chemotactic factor probably is a polar lipid. This chemotactic lipid may account, in part, for the leukocytic infiltration of the liver parenchyma that is observed during the course of acute alcoholic hepatitis. PMID:6434593

  1. Importance of extracellular divalent cations to polarisation of polymorphonuclear leukocytes induced by plasma.

    PubMed

    Harkin, D G; Bignold, L P

    1990-12-01

    The roles of extracellular calcium and magnesium ions in the polarisation of human peripheral blood polymorphonuclear leukocytes (PMN) induced by autologous fresh heparinised plasma were investigated by studying the effects of 5 mM chelators of divalent cations [ethylenediamine tetra-acetic acid (EDTA), ethylenebis-(oxyethylene-nitrilo)-tetra-acetic acid (EGTA) or disodium hydrogen citrate]. In addition, the effects of a blocker of membrane calcium channels (verapamil) were studied. Polarisation of PMN suspended in plasma (84.1 +/- 11.9%) was reduced by each chelating agent over 30 min (to 20.0 +/- 15.6% by EDTA, to 42.5 +/- 19.3% by EGTA and to 29.4 +/- 22.9% by citrate). Polarisation of PMN suspended in plasma treated with EDTA or EGTA was restored by inclusion of equimolar additional Ca2+ ions, and in plasma treated with EDTA, EGTA or citrate, by equimolar additional Mg2+ ions. Additional Mg2+ had no effect on the spherical shape of PMN in Hanks' solution and additional cations had no effects on the polarisation of PMN induced by fMLP. Cells rendered spherical by each chelating agent in plasma for 30 min retained their ability to polarise on addition of fMLP to the plasma-chelator medium. Verapamil (10(-4) M) markedly reduced polarisation in plasma (to 52 +/- 11.3%) but the same drug (10(-5) M) had no such effect. In contrast to the polarisation of cells in plasma, the polarisation response of PMN to N-formyl-methionyl-leucyl-phenylalanine (fMLP, 10(-8) M) in buffered Hanks' solution was not affected by any of the chelating agents or by verapamil, even in high concentration. These results indicate that extracellular divalent cations are necessary for the polarisation of PMN suspended in autologous plasma and that the mechanism of polarisation of PMN in plasma may be different to that of polarisation induced by fMLP. PMID:2128045

  2. Phagocytosis of virulent Porphyromonas gingivalis by human polymorphonuclear leukocytes requires specific immunoglobulin G.

    PubMed Central

    Cutler, C W; Kalmar, J R; Arnold, R R

    1991-01-01

    No studies to date clearly define the interactions between Porphyromonas gingivalis and human peripheral blood polymorphonuclear leukocytes (PMN), nor has a protective role for antibody to P. gingivalis been defined. Using a fluorochrome phagocytosis microassay, we investigated PMN phagocytosis and killing of P. gingivalis as a function of P. gingivalis-specific antibody. Sera from a nonimmune rabbit and a healthy human subject were not opsonic for virulent P. gingivalis A7436, W83, and HG405; phagocytosis of these strains (but not 33277) required opsonization with hyperimmune antiserum (RaPg). Diluting RaPg with a constant complement source decreased proportionally the number of P. gingivalis A7436 cells phagocytosed per phagocytic PMN. Enriching for the immunoglobulin G fraction of RAPg A7436 enriched for opsonic activity toward A7436. An opsonic evaluation of 18 serum samples from adult periodontitis patients revealed that only 3 adult periodontitis sera of 17 with elevated immunoglobulin G to P. gingivalis A7436 were opsonic for A7436 and, moreover, that the serum sample with the highest enzyme-linked immunosorbent assay titer was most opsonic (patient 1). However, the opsonic activity of serum from patient 1 was qualitatively and not just quantitatively different from that of the nonopsonic human sera (but was less effective opsonin than RaPg). Strain variability was observed in resistance of P. gingivalis to phagocytosis, and opsonization was strain specific for some, but not all, strains tested. An evaluation of killing of A7436 revealed that serum killing and extracellular killing of P. gingivalis were less effective alone when compared with intracellular PMN killing alone. PMID:2037370

  3. Activation of Polymorphonuclear Leukocytes by Candidate Biomaterials for an Implantable Glucose Sensor

    PubMed Central

    Sokolov, Andrey; Hellerud, Bernt Christian; Lambris, John D; Johannessen, Erik A; Mollnes, Tom Eirik

    2011-01-01

    Background Continuous monitoring of glucose by implantable microfabricated devices offers key advantages over current transcutaneous glucose sensors that limit usability due to their obtrusive nature and risk of infection. A successful sensory implant should be biocompatible and retain long-lasting function. Polymorphonuclear leukocytes (PMN) play a key role in the inflammatory system by releasing enzymes, cytokines, and reactive oxygen species, typically as a response to complement activation. The aim of this study was to perform an in vitro analysis of PMN activation as a marker for biocompatibility of materials and to evaluate the role of complement in the activation of PMN. Methods Fifteen candidate materials of an implantable glucose sensor were incubated in lepirudin-anticoagulated whole blood. The cluster of differentiation molecule 11b (CD11b) expression on PMN was analyzed with flow cytometry and the myeloperoxidase (MPO) concentration in plasma was analyzed with enzyme-linked immunosorbent assay. Complement activation was prevented by the C3 inhibitor compstatin or the C5 inhibitor eculizumab. Results Three of the biomaterials (cellulose ester, polyamide reverse osmosis membrane, and polyamide thin film membrane), all belonging to the membrane group, induced a substantial and significant increase in CD11b expression and MPO release. The changes were virtually identical for these two markers. Inhibition of complement with compstatin or eculizumab reduced the CD11b expression and MPO release dose dependently and in most cases back to baseline. The other 12 materials did not induce significant PMN activation. Conclusion Three of the 15 candidate materials triggered PMN activation in a complement-dependent manner and should therefore be avoided for implementation in implantable microsensors. PMID:22226271

  4. Vitamin E and vitamin C inhibit arachidonate-induced aggregation of human peripheral blood leukocytes in vitro

    Microsoft Academic Search

    S. Villa; A. Lorico; G. Morazzoni; G. de Gaetano; N. Semeraro

    1986-01-01

    Summary Arachidonate induces aggregation of human polymorphonuclear (PMN) and mononuclear (MNL) blood leukocytes. This is mediated by the lipoxygenase pathway, as it is prevented by lipoxygenase inhibitors and can also be induced by leukotriene B4 (LTB4). Vitamin E and vitamin C have profound effects on the functional state of leukocytes, some of which may involve the lipoxygenase pathway. This study

  5. Bordetella pertussis adenylate cyclase: effects of affinity-purified adenylate cyclase on human polymorphonuclear leukocyte functions.

    PubMed Central

    Friedman, R L; Fiederlein, R L; Glasser, L; Galgiani, J N

    1987-01-01

    Affinity-purified adenylate cyclase (AC) of Bordetella pertussis, free of contaminating pertussis toxin, was demonstrated to have biological effects on human polymorphonuclear leukocytes (PMN). AC at doses of 25 and 50 micrograms/ml increased intracellular cAMP levels in the phagocytes 7.6- to 23.5-fold, respectively, above basal levels. AC inhibited PMN chemiluminescence, chemotaxis, and superoxide production in a dose-dependent manner. The 50% inhibitory dose for chemotaxis and chemiluminescence was 36.5 micrograms/ml; for superoxide generation it was 71.0 micrograms/ml. Although these PMN metabolic functions were impaired, no effect on phagocytic activity was observed. Images PMID:2878884

  6. Microtechnique for Culturing Leukocytes from Whole Blood

    Microsoft Academic Search

    D. T. Arakaki; R. S. Sparkes

    1963-01-01

    By culturing heparinized whole blood, satisfactory chromosome preparations have been obtained easily and efficiently from blood samples as small as 0.05 ml. This technique eliminates any special treatment of the blood prior to initiating culture and minimizes the loss of leukocytes from the sample. It is particularly useful in those situations where separation of the leukocytes from other blood cells

  7. Polymorphonuclear Leukocyte Lysosomal Release in Response to Propionibacterium acne in Vitro and its Enhancement by Sera from Inflammatory Acne Patients

    Microsoft Academic Search

    G. F. Webster; J. J. Leyden; C. C. Tsai; P. Baehni; W. P. McArthur

    1980-01-01

    Propionibacteriunt acnes cells were tested for the ability to trigger lysosomal hydrolase release from human polymorphonuclear leukocytes. Representative strains of P. acnes serotype I and II failed to stimulate lysosomal release in the absence of serum. P. acnes growth culture supernatants failed to trigger release under any test condition. Addition of fresh or heat-inactivated human serum resulted in lysosomal hydrolase

  8. ORIGIN OF GRANULES IN POLYMORPHONUCLEAR LEUKOCYTES: Two Types Derived from Opposite Faces of the Golgi Complex in Developing Granulocytes

    Microsoft Academic Search

    DOROTHY FORD BAINTON; MARILYN G. FARQUHAR

    1966-01-01

    The origin, nature, and distribution of polymorphonuclear leukocyte (PMN) granules were invcstigated by examining developing granulocytcs from normal rabbit bone marrow which had been fixed in glutaraldchyde and postfixed in OsO4. Two distinct types of granules, azurophil and specific, were distinguished on the basis of their differences in size, density, and time and modc of origin. Both types are produced

  9. ?2 integrins (CD11/18) are essential for the chemosensory adhesion and migration of polymorphonuclear leukocytes on bacterial cellulose.

    PubMed

    Kim, Gun-Dong; Lee, Seung Eun; Yang, Hana; Park, Hye Rim; Son, Gun Woo; Park, Cheung-Seog; Park, Yong Seek

    2015-05-01

    Bacterial cellulose (BC) has been studied widely for applications in biomedical materials such as prosthetic artificial blood vessels owing to its unique characteristics, which include nontoxicity and nonimmunogenicity as compared with synthetic biopolymers such as expanded polytetrafluorethylene (ePTFE). However, to date, studies on the relative effect of leukocytes on BC as a prosthetic vascular graft are insufficient. Polymorphonuclear leukocytes (PMN) play a pivotal role in early-phase immune response to bacterial or periprosthetic infection. PMN recruitment at sites of infection or inflammation mediated by various integrins such as ?2 integrin family (CD11/CD18 family). Therefore, we discuss our investigations into the mechanisms by which ?2 integrins-mediated chemosensory adhesion and migration of PMN on the vascular graft surface, BC. Our results show that CD11b/CD18 components mainly mediate PMN adherence on BC. CD11b/CD18 displays weak coordination with the other two ? subunits (CD11a and CD11c). Furthermore, it was found that the ? subunit (CD18) plays a critical role in both the adhesion and migration of N-formylmethionyl-leucyl-phenylalanine (fMLP)-stimulated PMN on BC. The activity of CD18 contrasts with that of the individual ? subunits. Among these, only CD11b displayed inhibition of PMN migration on BC surfaces. PMID:25231265

  10. CD45 modulation of CXCR1 and CXCR2 in human polymorphonuclear leukocytes.

    PubMed

    Mitchell, G B; Khandaker, M H; Rahimpour, R; Xu, L; Lazarovits, A I; Pickering, J G; Suria, H; Madrenas, J; Pomerantz, D K; Feldman, R D; Kelvin, D J

    1999-05-01

    All leukocytes express the cell surface glycoprotein CD45, which has intrinsic intracellular protein tyrosine phosphatase activity. CD45 is known to play a regulatory role in activation-induced signaling in lymphocytes; however, little is known of its role in non-lymphoid leukocytes. Therefore, we examined the potential effect of CD45 on chemokine-induced signaling in human neutrophils (polymorphonuclear cells, PMN). Treating isolated PMN for 2 h with an anti-CD45RB antibody (Bra11) down-modulated expression of the chemokine receptors CXCR1 and CXCR2 to 44 +/- 10% and 47 +/- 9% of their respective controls. The tyrosine kinase inhibitors genistein and herbimycin A significantly inhibited the Bra11-induced down-modulation of CXCR1 and CXCR2. Furthermore, Bra11-treated PMN were functionally inhibited in their capacity to exhibit IL-8-induced transient intracellular Ca2+ increases. Selected targeting of CXC receptors is indicated by the fact that N-formyl-Met-Leu-Phe (fMLP) receptor expression and function were not lost following Bra11 treatment. The effect of Bra11 on IL-8-mediated function and receptor expression was paralleled by decreased tyrosine phosphorylation of a 54- to 60-kDa protein. These findings indicate that CD45 can act to modulate PMN responses to chemokines; thus agents regulating CD45 can potentially modulate leukocyte traffic and may represent a novel therapeutic approach towards the treatment of inflammatory diseases. PMID:10359100

  11. Determination of phagocytosis of /sup 32/P-labeled Staphylococcus aureus by bovine polymorphonuclear leukocytes

    SciTech Connect

    Dulin, A.M.; Paape, M.J.; Weinland, B.T.

    1984-04-01

    A procedure for the measurement of phagocytosis by bovine polymorphonuclear leukocytes (PMN) of /sup 32/P-labeled Staphylococcus aureus was modified so that a larger number of samples could be compared in a single run, and smaller volumes of sample, PMN, and /sup 32/P-labeled S aureus could be used. Results were highly reproducible, with a coefficient of variation between duplicate determinations of less than or equal to 2%. Lysostaphin was prepared from the supernatant of S staphylolyticus and was compared with a commercially available preparation. Effects of lysostaphin on PMN and influence of incubation media on release of /sup 32/P from /sup 32/P-labeled S aureus by lysostaphin were examined.

  12. Determination of phagocytosis of 32P-labeled Staphylococcus aureus by bovine polymorphonuclear leukocytes.

    PubMed

    Dulin, A M; Paape, M J; Weinland, B T

    1984-04-01

    A procedure for the measurement of phagocytosis by bovine polymorphonuclear leukocytes (PMN) of 32P-labeled Staphylococcus aureus was modified so that a larger number of samples could be compared in a single run, and smaller volumes of sample, PMN, and 32P-labeled S aureus could be used. Results were highly reproducible, with a coefficient of variation between duplicate determinations of less than or equal to 2%. Lysostaphin was prepared from the supernatant of S staphylolyticus and was compared with a commercially available preparation. Effects of lysostaphin on PMN and influence of incubation media on release of 32P from 32P-labeled S aureus by lysostaphin were examined. PMID:6731995

  13. THE INTERACTION IN VITRO BETWEEN GROUP B MENINGOCOCCI AND RABBIT POLYMORPHONUCLEAR LEUKOCYTES

    PubMed Central

    Roberts, Richard B.

    1967-01-01

    The interaction in vitro between group B meningococci and rabbit polymorphonuclear leukocytes has been described. Phagocytosis did not occur in the presence of normal rabbit serum. Antiserum collected 12–21 days following one subcutaneous inoculation of living log phase meningococci exhibited opsonic activity with type specificity; this opsonic action depended on both heat-labile and heat-stable factors. Following ingestion by granulocytes, meningococci were rapidly killed. These studies suggest that group B meningococcal strains contain specific antiphagocytic surface factors of an as yet unknown chemical nature. Antisera obtained 4 or more wk after immunization showed bactericidal activity with the same type specificity as opsonic activity. This bactericidal activity was also lost after heating and restored by the addition of normal serum. Further studies on opsonins and bactericidins for meningococci may shed light on virulence factors in these microorganisms, and may prove useful for a more precise classification of meningococci according to type rather than group specificity. PMID:4964859

  14. Membrane cholesterol modulates the fluid shear stress response of polymorphonuclear leukocytes via its effects on membrane fluidity.

    PubMed

    Zhang, Xiaoyan; Hurng, Jonathan; Rateri, Debra L; Daugherty, Alan; Schmid-Schönbein, Geert W; Shin, Hainsworth Y

    2011-08-01

    Continuous exposure of polymorphonuclear leukocytes (PMNLs) to circulatory hemodynamics points to fluid flow as a biophysical regulator of their activity. Specifically, fluid flow-derived shear stresses deactivate leukocytes via actions on the conformational activities of proteins on the cell surface. Because membrane properties affect activities of membrane-bound proteins, we hypothesized that changes in the physical properties of cell membranes influence PMNL sensitivity to fluid shear stress. For this purpose, we modified PMNL membranes and showed that the cellular mechanosensitivity to shear was impaired whether we increased, reduced, or disrupted the organization of cholesterol within the lipid bilayer. Notably, PMNLs with enriched membrane cholesterol exhibited attenuated pseudopod retraction responses to shear that were recovered by select concentrations of benzyl alcohol (a membrane fluidizer). In fact, PMNL responses to shear positively correlated (R(2) = 0.96; P < 0.0001) with cholesterol-related membrane fluidity. Moreover, in low-density lipoprotein receptor-deficient (LDLr(-/-)) mice fed a high-fat diet (a hypercholesterolemia model), PMNL shear-responses correlated (R(2) = 0.5; P < 0.01) with blood concentrations of unesterified (i.e., free) cholesterol. In this regard, the shear-responses of PMNLs gradually diminished and eventually reversed as free cholesterol levels in blood increased during 8 wk of the high-fat diet. Collectively, our results provided evidence that cholesterol is an important component of the PMNL mechanotransducing capacity and elevated membrane cholesterol impairs PMNL shear-responses at least partially through its impact on membrane fluidity. This cholesterol-linked perturbation may contribute to dysregulated PMNL activity (e.g., chronic inflammation) related to hypercholesterolemia and causal for cardiovascular pathologies (e.g., atherosclerosis). PMID:21525434

  15. Membrane cholesterol modulates the fluid shear stress response of polymorphonuclear leukocytes via its effects on membrane fluidity

    PubMed Central

    Zhang, Xiaoyan; Hurng, Jonathan; Rateri, Debra L.; Daugherty, Alan; Schmid-Schönbein, Geert W.

    2011-01-01

    Continuous exposure of polymorphonuclear leukocytes (PMNLs) to circulatory hemodynamics points to fluid flow as a biophysical regulator of their activity. Specifically, fluid flow-derived shear stresses deactivate leukocytes via actions on the conformational activities of proteins on the cell surface. Because membrane properties affect activities of membrane-bound proteins, we hypothesized that changes in the physical properties of cell membranes influence PMNL sensitivity to fluid shear stress. For this purpose, we modified PMNL membranes and showed that the cellular mechanosensitivity to shear was impaired whether we increased, reduced, or disrupted the organization of cholesterol within the lipid bilayer. Notably, PMNLs with enriched membrane cholesterol exhibited attenuated pseudopod retraction responses to shear that were recovered by select concentrations of benzyl alcohol (a membrane fluidizer). In fact, PMNL responses to shear positively correlated (R2 = 0.96; P < 0.0001) with cholesterol-related membrane fluidity. Moreover, in low-density lipoprotein receptor-deficient (LDLr?/?) mice fed a high-fat diet (a hypercholesterolemia model), PMNL shear-responses correlated (R2 = 0.5; P < 0.01) with blood concentrations of unesterified (i.e., free) cholesterol. In this regard, the shear-responses of PMNLs gradually diminished and eventually reversed as free cholesterol levels in blood increased during 8 wk of the high-fat diet. Collectively, our results provided evidence that cholesterol is an important component of the PMNL mechanotransducing capacity and elevated membrane cholesterol impairs PMNL shear-responses at least partially through its impact on membrane fluidity. This cholesterol-linked perturbation may contribute to dysregulated PMNL activity (e.g., chronic inflammation) related to hypercholesterolemia and causal for cardiovascular pathologies (e.g., atherosclerosis). PMID:21525434

  16. Stimulatory effect of some plant extracts used in homeopathy on the phagocytosis induced chemiluminescence of polymorphonuclear leukocytes.

    PubMed

    Crocnan, D O; Greabu, M; Olinescu, R

    2000-01-01

    Some plant extracts on a large range of dilutions as used in Homeopathy were tested on the chemiluminescence emission produced by polymorphonuclear leukocytes. The high stimulatory action was noticed when extracts from Uvae Ursi and Saponaria were tested, as the classical effect exerted by zymosan was exceeded. A moderate stimulatory action comparable with that of zymosan was found when extracts from Echmaceea, Aleo and Prumis were used, as well as in the case of Propolis. The relationship between stimulatory effect and the concentration range is modulated as function of the extract source, several peaks being observed for some dilutions (Saponana), but generally no quantitative relations were obtained. By studying the time when a chemiluminescence peak was observed, it is possible to estimate wether the weight of the NADPH oxidase or myeloperoxidase pathways are involved in the stimulatory effect on polymorphonuclear leukocytes. PMID:11712436

  17. A Ca2+-insensitive form of fura-2 associated with polymorphonuclear leukocytes. Assessment and accurate Ca2+ measurement

    Microsoft Academic Search

    Mary Scanlon; David A. Williams; Fredric S. Fay

    1987-01-01

    The new, fluorescent Ca2+ indicator, fura-2, promises to expand our understanding of the role of subcellular changes in Ca2+ underlying cell function. During an investigation of the role of Ca2+ in the polarization response of human polymorphonuclear leukocytes to formyl-methionyl-leucyl-phenylalanine, we found that fura-2 trapped by cells incubated with the acetoxy-methyl ester of fura-2, F2-AM, yielded measurements of Ca2+ that

  18. Polymorphonuclear leukocyte membrane fluidity and cytosolic Ca2+ content in different clinical conditions.

    PubMed

    Caimi, G; Canino, B; Montana, M; Ventimiglia, G; Catania, A; Lo Presti, R

    1997-01-01

    The aim of the study was to evaluate the polymorphonuclear leukocyte (PMN) membrane fluidity and PMN cytosolic Ca2+ content in several clinical conditions: diabetes mellitus, vascular atherosclerotic disease (VAD), chronic renal failure (CRF), essential hypertension (EH). In 13 subjects with insulin-dependent diabetes mellitus (IDDM), in 24 subjects with non-insulin-dependent diabetes mellitus (NIDDM), in 42 VAD subjects, in 23 VAD subjects with NIDDM, in 15 subjects with CRF and in 12 subjects with EH, we determined the PMN membrane fluidity, obtained marking unstimulated PMN cells with fluorescent probe 1-[4-(trimethylamino)phenyl]-6-phenyl-1,3,5-hexatriene (TMA-DPH), and considering the fluorescence polarization degree, and the PMN cytosolic Ca2+ content, obtained marking unstimulated PMN cells with the fluorescent probe Fura2-AM and considering the ratio between the Fura2-Ca2+ complex and the unchelated Fura 2 fluorescence intensity. From the obtained data it is evident that PMN membrane fluidity does not distinguish normals from IDDM subjects, NIDDM subjects, VAD subjects with and without NIDDM, CRF subjects and hypertensives. PMN cytosolic Ca2+ content, in comparison with normal controls, is significantly increased in VAD subjects (p < 0.01), in VAD subjects with NIDDM (p < 0.001), in CRF subjects (p < 0.001) and in hypertensives (p < 0.05). No correlation was found between PMN membrane fluidity and PMN cytosolic Ca2+ content. The study of these PMN parameters can be useful in the understanding of the role of leukocytes in the vascular damage that characterizes these clinical conditions. PMID:9356786

  19. Effect of mechanical deformation on structure and function of polymorphonuclear leukocytes.

    PubMed

    Kitagawa, Y; Van Eeden, S F; Redenbach, D M; Daya, M; Walker, B A; Klut, M E; Wiggs, B R; Hogg, J C

    1997-05-01

    The present studies were designed to test the hypothesis that mechanical deformation of polymorphonuclear leukocytes (PMN) leads to functional changes that might influence their transit in the pulmonary capillaries. Human leukocytes were passed through 5- or 3-micron-pore polycarbonate filters under controlled conditions. Morphometric analysis showed that the majority of PMN were deformed and that this deformation persisted longer after filtration through 3-micron filters than through 5-micron filters (P < 0.05) but did not result in the cytoskeletal polarization characteristic of migrating cells. Flow cytometric studies of the filtered PMN showed that there was a transient increase in the cytosolic free Ca2+ concentration after both 3- and 5-micron filtration (P < 0.01) with an increase in F-actin content after 3-micron filtration (P < 0.05). Although L-selectin expression on PMN was not changed by either 5- or 3-micron filtration, CD18 and CD11b were increased by 3-micron filtration (P < 0.05). Priming of the PMN with N-formyl-methionyl-leucyl-phenylalanine (0.5 nM) before filtration resulted in an increase of CD11b by both 5 (P < 0.05)- and 3-micron (P < 0.01) filtration. Neither 5- nor 3-micron filtration induced hydrogen peroxide production. We conclude that mechanical deformation of PMN, similar to what occurs in the pulmonary microvessels, induces both structural and functional changes in the cells, which might influence their passage through the pulmonary capillary bed. PMID:9134884

  20. Interaction of polymorphonuclear leukocytes with smooth and rough strains of Brucella abortus.

    PubMed Central

    Kreutzer, D L; Dreyfus, L A; Robertson, D C

    1979-01-01

    The bactericidal activity of guinea pig and human polymorphonuclear leukocytes (PMNs) against a smooth-intermediate strain (45/0) and a rough strain (45/20) of Brucella abortus has been examined. After incubation for 120 min, guinea pig PMNs incubated with either the smooth strain 45/0 or the rough strain 45/20 exhibited no bactericidal activity against the former and caused only a 34% decrease in viability of the latter. Human PMNs were more bactericidal than guinea pig PMNs to both strains; however, the killing of strain 45/20 by human PMNs was less than that observed in control experiments with S. aureus strain 502A. Both strains of B. abortus readily associated with guinea pig and human PMNs, and the bacteria were apparently ingested without stimulation of the hexose monophosphate pathway. Lysates (10 micrograms/ml, pH 5.5), prepared from guinea pig or human granules, were not particularly toxic to either strain unless supplemented with H2O2 and a halide (I- or Cl-). An oxygen-dependent killing system appeared to be lethal against both strains of B. abortus, with I- being more active than Cl- in the presence of H2O2 and granule lysate. The data suggest that degranulation after ingestion of Brucella by phagocytes does not occur due to the lack of a proper stimulus or possibly the baccilli actively inhibit the degranulation process thereby protecting the microbe from killing systems normally effective against extracellular parasites. PMID:110680

  1. Altered Ca2+ homeostasis in polymorphonuclear leukocytes from chronic myeloid leukaemia patients

    PubMed Central

    Revankar, Chetana M; Advani, Suresh H; Naik, Nishigandha R

    2006-01-01

    Background In polymorphonuclear leukocytes (PMNL), mobilization of calcium ions is one of the early events triggered by binding of chemoattractant to its receptors. Besides chemotaxis, a variety of other functional responses are dependent on calcium ion mobilization. PMNL from chronic myeloid leukaemia (CML) patients that were morphologically indistinguishable from normal PMNL were found to be defective in various functions stimulated by a chemoattractant – fMLP. To study the mechanism underlying defective functions in CML PMNL, we studied calcium mobilization in CML PMNL in response to two different classical chemoattractants, fMLP and C5a. Results Release of calcium estimated by flow cytometry and spectrofluorimetry using fluo-3 as an indicator showed that the [Ca2+]i levels were lower in CML PMNL as compared to those in normal PMNL. But, both normal and CML PMNL showed maximum [Ca2+]i in response to fMLP and C5a at 10 sec and 30 sec, respectively. Spectrofluorimetric analysis of the total calcium release in chemoattractant treated PMNL indicated more and faster efflux of [Ca2+]i in CML PMNL as compared to normal PMNL. Conclusion Fine-tuning of Ca2+ homeostasis was altered in CML PMNL. The altered Ca2+ homeostasis may contribute to the defective functions of CML PMNL. PMID:17129370

  2. Boomerang effect between [Met]-enkephalin derivatives and human polymorphonuclear leukocytes.

    PubMed

    Rabgaoui, N; Slaoui-Hasnaoui, A; Torreilles, J

    1993-05-01

    [Met]-enkephalin or its precursor, pre-[Met]-enkephalin, were exposed to activated oxygen species produced by human phorbol myristate acetate (PMA)-stimulated polymorphonuclear leukocytes (PMNs) and then analyzed by high-pressure liquid chromatography (HPLC). The chromatograms recorded at the tyrosine maximum wavelength (lambda em 300 nm and lambda ex 280 nm) showed the formation of new peptides by oxidation of methionyl residue in position 5 and ortho, meta, or para hydroxylation of phenylalanyl residue in position 4. The chromatograms recorded at the dityrosine maximum wavelength (lambda em 400 nm and lambda ex 325 nm) showed the formation of new dimeric peptides which contained two [Met]-enkephalin-derivatives linked by a dityrosyl group. These new peptides were tested for chemiluminescence response to PMA-stimulated PMNs. [Met]-enkephalin, pre-[Met]-enkephalin, and the methionyl-oxidized derivatives suppressed the PMA-induced respiratory burst of PMNs. Conversely, after hydroxylation by activated oxygen species released by stimulated PMNs, these peptides enhanced the PMA-induced respiratory burst of PMNs. In the same conditions, dimeric peptides had no effect. PMID:8349141

  3. Effects of polymorphonuclear leukocyte transmigration on the barrier function of cultured intestinal epithelial monolayers.

    PubMed Central

    Nash, S; Stafford, J; Madara, J L

    1987-01-01

    We describe a model to study the effects of polymorphonuclear leukocyte (PMN) transmigration on the intestinal epithelial barrier. Human PMN were induced to transmigrate across high resistance monolayers of a cultured human intestinal epithelial cell line (T84 cells) by chemotactic gradients produced by formyl methionyl leucyl phenylalanine (FMLP). With maximal transmigration monolayer resistance decreased by 48 +/- 12.6% in 15 min and by 83 +/- 1.6% in 60 min. This response was dependent on the size of the FMLP gradient and the density of PMN transmigration. The decrease in resistance correlated with number of PMN migrating across monolayers, and was accompanied by increases in flux of paracellular tracers. Macromolecular tracer studies localized the leak sites to foci at which PMN impaled the epithelium. Removal of the chemotactic gradient led to restoration of baseline resistance within 18 h. PMN transmigration across intestinal epithelial monolayers occurs via intercellular occluding junctions and may be associated with a reversible increase in epithelial permeability. Images PMID:3116044

  4. The essential oil of bergamot stimulates reactive oxygen species production in human polymorphonuclear leukocytes.

    PubMed

    Cosentino, Marco; Luini, Alessandra; Bombelli, Raffaella; Corasaniti, Maria T; Bagetta, Giacinto; Marino, Franca

    2014-08-01

    Bergamot (Citrus aurantium L. subsp. bergamia) essential oil (BEO) is used in folk medicine as an antiseptic and anthelminthic and to facilitate wound healing. Evidence indicates that BEO has substantial antimicrobial activity; however its effects on immunity have never been examined. We studied the effects of BEO on reactive oxygen species (ROS) production in human polymorphonuclear leukocytes (PMN) and the role of Ca(2+) in the functional responses evoked by BEO in these cells. Results show that BEO increased intracellular ROS production in human PMN, an effect that required the contribution of extracellular (and, to a lesser extent, of intracellular) Ca(2+) . Bergamot essential oil also significantly increased ROS production induced by the chemotactic peptide N-formyl-Met-Leu-Phe and reduced the response to the protein kinase C activator phorbol myristate acetate. In conclusion, this is the first report showing the ability of BEO to increase ROS production in human PMN. This effect could both contribute to the activity of BEO in infections and in tissue healing as well as underlie an intrinsic proinflammatory potential. The relevance of these findings for the clinical uses of BEO needs careful consideration. PMID:24458921

  5. Staphylococcal serine proteinase increases intracellular free calcium concentration in human polymorphonuclear leukocytes.

    PubMed

    Nowak, D; Krol, M; Miedzobrodzki, J; Bialasiewicz, P

    1997-10-01

    Staphylococcal serine proteinase (SSP) can influence various functions of human polymorphonuclear leukocytes (PMNL) including chemotaxis and phagocytosis. Since the rise in intracellular free calcium concentration is an important step in signal transduction leading to phagocyte activation, we tested the ability of SSP to increase the intracellular free calcium concentration in human PMNL using the fluorescent calcium indicator Fura-2AM. PMNL isolated from healthy donors responded to SSP in the concentration range of 10 to 100 micrograms/ml. The highest Ca2+ rise (104 +/- 47 nM) was observed for 10 micrograms/ml SSP. It was mainly dependent (81 +/- 11%) on extracellular calcium influx, however, SSP mobilized 68 +/- 7% of Ca2+ from intracellular calcium stores. Boiling of SSP or preincubation with phenylmethylsulphonylfluoride (an serine proteinase inhibitor) did not change its ability to increase intracellular free calcium concentration in PMNL. It suggests that active center of SSP is not responsible for Ca2+ mobilization. Finally, PMNL responded to each of three consecutive stimulations with SSP independently of the presence of high or low extracellular Ca2+ concentration. This may be an additional mechanism responsible for activation of human PMNL and degradation of alveolar walls during the staphylococcal infection in the lower airways. PMID:9403110

  6. Yersinia pestis Type III Secretion System-Dependent Inhibition of Human Polymorphonuclear Leukocyte Function?

    PubMed Central

    Spinner, Justin L.; Cundiff, Jennifer A.; Kobayashi, Scott D.

    2008-01-01

    Human polymorphonuclear leukocytes (PMNs, or neutrophils) are the primary innate host defense against invading bacterial pathogens. Neutrophils are rapidly recruited to sites of infection and ingest microorganisms through a process known as phagocytosis. Following phagocytosis by human PMNs, microorganisms are killed by reactive oxygen species (ROS) and microbicidal products contained within granules. Yersinia pestis, the causative agent of plague, is capable of rapid replication and dissemination from sites of infection in the host. Although Y. pestis survives in macrophages, the bacterial fate following interaction with human PMNs is less clear. The ability of Y. pestis to inhibit phagocytosis by human PMNs was assessed by differential fluorescence microscopy and was shown to be dependent on expression of the type III secretion system (TTSS). Previous studies have demonstrated that TTSS expression in enteropathogenic Yersinia spp. also inhibits the respiratory burst in PMNs and macrophages, and we show here that human PMN ROS production is similarly repressed by Y. pestis. However, exclusion of uningested TTSS-expressing Y. pestis with gentamicin revealed that intracellular bacteria are eliminated by human PMNs, similar to bacteria lacking the TTSS. In summary, our results suggest that the Y. pestis TTSS contributes to extracellular survival following interactions with human PMNs and that the intracellular fate is independent of TTSS inhibition of neutrophil ROS production. PMID:18490459

  7. Delivery of rifampicin-chitin nanoparticles into the intracellular compartment of polymorphonuclear leukocytes.

    PubMed

    Smitha, K T; Nisha, N; Maya, S; Biswas, Raja; Jayakumar, R

    2015-03-01

    Polymorphonuclear leukocytes (PMNs) provide the primary host defence against invading pathogens by producing reactive oxygen species (ROS) and microbicidal products. However, few pathogens can survive for a prolonged period of time within the PMNs. Additionally their intracellular lifestyle within the PMNs protect themselves from the additional lethal action of host immune systems such as antibodies and complements. Antibiotic delivery into the intracellular compartments of PMNs is a major challenge in the field of infectious diseases. In order to deliver antibiotics within the PMNs and for the better treatment of intracellular bacterial infections we synthesized rifampicin (RIF) loaded amorphous chitin nanoparticles (RIF-ACNPs) of 350±50 nm in diameter. RIF-ACNPs nanoparticles are found to be non-hemolytic and non-toxic against a variety of host cells. The release of rifampicin from the prepared nanoparticles was ?60% in 24 h, followed by a sustained pattern till 72 h. The RIF-ACNPs nanoparticles showed 5-6 fold enhanced delivery of RIF into the intracellular compartments of PMNs. The RIF-ACNPs showed anti-microbial activity against Escherichia coli, Staphylococcus aureus and a variety of other bacteria. In summary, our results suggest that RIF-ACNPs could be used to treat a variety of intracellular bacterial infections. PMID:25475841

  8. Plasma markers of platelet and polymorphonuclear leukocyte activation in young adults with acute myocardial infarction.

    PubMed

    Caimi, Gregorio; Hoffmann, Enrico; Montana, Maria; Incalcaterra, Egle; Dispensa, Flavia; D'Amico, Teresa; Canino, Baldassare; Lo Presti, Rosalia

    2005-01-01

    Our goal was to evaluate some plasma markers of platelet and polymorphonuclear leukocyte (PMN) activation in a group of young adults with acute myocardial infarction (AMI) at the initial stage and after three months. We enrolled 49 AMI subjects aged<45 years and examined plasmatic levels of platelet factor 4 (PF4), beta-thromboglobulin (beta-TG), elastase and myeloperoxidase (MPO) using ELISA methods. PF4 and beta-TG were increased, compared to control subjects, both at the initial stage and after 3 months. In control subjects and in AMI patients, at both times of observation, there was a significant and positive correlation between the two platelet parameters, while no correlation was present between each parameter and platelet count. In AMI patients there was an increase in elastase levels in comparison with the control group; this increase was evident at the initial stage and after 3 months. There was no difference in MPO levels between control subjects and AMI patients. In control subjects and in AMI patients there was a significant and positive correlation between elastase and MPO level, whereas no relationship was found between each marker and PMN count. Our data show that in young AMI patients the discharge treatment including antiplatelet drugs did not modify platelet activation and suggest the association of molecules able to inhibit PMN activation to the conventional therapy of these AMI patients. PMID:15665428

  9. Caffeic Acid Phenethyl Ester and Its Amide Analogue Are Potent Inhibitors of Leukotriene Biosynthesis in Human Polymorphonuclear Leukocytes

    PubMed Central

    Boudreau, Luc H.; Maillet, Jacques; LeBlanc, Luc M.; Jean-François, Jacques; Touaibia, Mohamed; Flamand, Nicolas; Surette, Marc E.

    2012-01-01

    Background 5-lipoxygenase (5-LO) catalyses the transformation of arachidonic acid (AA) into leukotrienes (LTs), which are important lipid mediators of inflammation. LTs have been directly implicated in inflammatory diseases like asthma, atherosclerosis and rheumatoid arthritis; therefore inhibition of LT biosynthesis is a strategy for the treatment of these chronic diseases. Methodology/Principal Findings Analogues of caffeic acid, including the naturally-occurring caffeic acid phenethyl ester (CAPE), were synthesized and evaluated for their capacity to inhibit 5-LO and LTs biosynthesis in human polymorphonuclear leukocytes (PMNL) and whole blood. Anti-free radical and anti-oxidant activities of the compounds were also measured. Caffeic acid did not inhibit 5-LO activity or LT biosynthesis at concentrations up to 10 µM. CAPE inhibited 5-LO activity (IC50 0.13 µM, 95% CI 0.08–0.23 µM) more effectively than the clinically-approved 5-LO inhibitor zileuton (IC50 3.5 µM, 95% CI 2.3–5.4 µM). CAPE was also more effective than zileuton for the inhibition of LT biosynthesis in PMNL but the compounds were equipotent in whole blood. The activity of the amide analogue of CAPE was similar to that of zileuton. Inhibition of LT biosynthesis by CAPE was the result of the inhibition of 5-LO and of AA release. Caffeic acid, CAPE and its amide analog were free radical scavengers and antioxidants with IC50 values in the low µM range; however, the phenethyl moiety of CAPE was required for effective inhibition of 5-LO and LT biosynthesis. Conclusions CAPE is a potent LT biosynthesis inhibitor that blocks 5-LO activity and AA release. The CAPE structure can be used as a framework for the rational design of stable and potent inhibitors of LT biosynthesis. PMID:22347509

  10. Polymorphonuclear leukocytes restrict growth of Pseudomonas aeruginosa in the lungs of cystic fibrosis patients.

    PubMed

    Kragh, Kasper N; Alhede, Morten; Jensen, Peter Ř; Moser, Claus; Scheike, Thomas; Jacobsen, Carsten S; Seier Poulsen, Steen; Eickhardt-Sřrensen, Steffen Robert; Trřstrup, Hannah; Christoffersen, Lars; Hougen, Hans-Petter; Rickelt, Lars F; Kühl, Michael; Hřiby, Niels; Bjarnsholt, Thomas

    2014-11-01

    Cystic fibrosis (CF) patients have increased susceptibility to chronic lung infections by Pseudomonas aeruginosa, but the ecophysiology within the CF lung during infections is poorly understood. The aim of this study was to elucidate the in vivo growth physiology of P. aeruginosa within lungs of chronically infected CF patients. A novel, quantitative peptide nucleic acid (PNA) fluorescence in situ hybridization (PNA-FISH)-based method was used to estimate the in vivo growth rates of P. aeruginosa directly in lung tissue samples from CF patients and the growth rates of P. aeruginosa in infected lungs in a mouse model. The growth rate of P. aeruginosa within CF lungs did not correlate with the dimensions of bacterial aggregates but showed an inverse correlation to the concentration of polymorphonuclear leukocytes (PMNs) surrounding the bacteria. A growth-limiting effect on P. aeruginosa by PMNs was also observed in vitro, where this limitation was alleviated in the presence of the alternative electron acceptor nitrate. The finding that P. aeruginosa growth patterns correlate with the number of surrounding PMNs points to a bacteriostatic effect by PMNs via their strong O2 consumption, which slows the growth of P. aeruginosa in infected CF lungs. In support of this, the growth of P. aeruginosa was significantly higher in the respiratory airways than in the conducting airways of mice. These results indicate a complex host-pathogen interaction in chronic P. aeruginosa infection of the CF lung whereby PMNs slow the growth of the bacteria and render them less susceptible to antibiotic treatment while enabling them to persist by anaerobic respiration. PMID:25114118

  11. Role of microtubule assembly in lysosomal enzyme secretion from human polymorphonuclear leukocytes. A reevaluation

    PubMed Central

    1977-01-01

    The dose-related inhibition by colchicine of both lysosomal enzyme release and microtubule assembly was studied in human polymorphonuclear leukocytes (PMN) exposed to the nonphagocytic stimulus, zymosan-treated serum (ZTS). Cells were pretreated with colchicine (60 min, 37 degrees C) with or without cytochalasin B (5 microng/ml, 10 min) and then stimulated with ZTS (10%). Microtubule numbers in both cytochalasin B- treated and untreated PMN were increased by stimulation and depressed below resting levels in a dose-response fashion by colchicine concentrations above 10(-7) M. These concentrations also inhibited enzyme release in a dose-response fashion although the inhibition of microtubule assembly was proportionately greater than the inhibition of enzyme release. Other aspects of PMN morphology were affected by colchicine. Cytochalasin B-treated PMN were rounded, and in thin sections the retracted plasma membrane appeared as invaginations oriented toward centrally located centrioles. Membrane invaginations were restricted to the cell periphery in cells treated with inhibitory concentrations of colchicine, and the centrioles and Golgi apparatus were displaced from their usual position. After stimulation and subsequent degranulation, the size and number of membrane invaginations greatly increased. They remained peripheral in cells pretreated with greater than 10(-7) M colchicine but were numerous in the pericentriolar region in cells treated with less than 10(-7) M. Similarly, untreated PMN that were permitted to phagocytose immune precipitates had many phagosomes adjacent to the centriole. After colchicine treatment, phagosomes were distributed randomly, without any preferential association with the centrioles. These data suggest that microtubules are involved in maintaining the internal organization of cells and the topologic relationships between organelles and the plasma membrane. PMID:856834

  12. Increased activity of 5-lipoxygenase in polymorphonuclear leukocytes from asthmatic patients

    SciTech Connect

    Mita, H.; Yui, Y.; Taniguchi, N.; Yasueda, H.; Shida, T.

    1985-09-09

    The formation of 5-lipoxygenase products of arachidonic acid, 5-HETE and 5,12-diHETE, was determined in 100,000 x g supernatant of polymorphonuclear leukocytes from 17 healthy subjects, 17 patients with extrinsic asthma and 15 patients with intrinsic asthma. After the supernatant was incubated with /sup 14/C-arachidonic acid in the presence of calcium and indomethacin, the lipoxygenase products of arachidonic acid were separated by thin layer chromatography. The results were expressed as the percentage conversion of /sup 14/C-arachidonic acid into the product per 10/sup 7/ cells. The formation of 5,12-diHETE, but not of the 5-HETE, was significantly increased in the cells from the group of patients with extrinsic asthma (4.38 +/- 0.78%, mean +/- S.E.; p < 0.01) and intrinsic asthma (6.09 +/- 1.11%; p < 0.01), when compared to normal subjects (1.74 +/- 0.30%). Both extrinsic and intrinsic asthmatics had significantly enhanced 5-lipoxygenase activity, which was expressed as the sum of percentage conversion of /sup 14/C-arachidonic acid into 5-HETE and 5,12-diHETE. The percentage conversion in normal subjects was 4.19 +/- 0.39%, 6.24 +/- 0.84% for 17 patients with extrinsic asthma (p < 0.05), and 8.59 +/- 1.29% for 15 patients with intrinsic asthma (p < 0.01). There was no significant difference between these asthmatic groups. These results indicate that 5-lipoxygenase activity is increased in patients with bronchial asthma. 22 references, 3 figures.

  13. Antipseudomonal agents exhibit differential pharmacodynamic interactions with human polymorphonuclear leukocytes against established biofilms of Pseudomonas aeruginosa.

    PubMed

    Chatzimoschou, Athanasios; Simitsopoulou, Maria; Antachopoulos, Charalampos; Walsh, Thomas J; Roilides, Emmanuel

    2015-04-01

    Pseudomonas aeruginosa is the most common pathogen infecting the lower respiratory tract of cystic fibrosis (CF) patients, where it forms tracheobronchial biofilms. Pseudomonas biofilms are refractory to antibacterials and to phagocytic cells with innate immunity, leading to refractory infection. Little is known about the interaction between antipseudomonal agents and phagocytic cells in eradication of P. aeruginosa biofilms. Herein, we investigated the capacity of three antipseudomonal agents, amikacin (AMK), ceftazidime (CAZ), and ciprofloxacin (CIP), to interact with human polymorphonuclear leukocytes (PMNs) against biofilms and planktonic cells of P. aeruginosa isolates recovered from sputa of CF patients. Three of the isolates were resistant and three were susceptible to each of these antibiotics. The concentrations studied (2, 8, and 32 mg/liter) were subinhibitory for biofilms of resistant isolates, whereas for biofilms of susceptible isolates, they ranged between sub-MIC and 2 × MIC values. The activity of each antibiotic alone or in combination with human PMNs against 48-h mature biofilms or planktonic cells was determined by XTT [2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide] assay. All combinations of AMK with PMNs resulted in synergistic or additive effects against planktonic cells and biofilms of P. aeruginosa isolates compared to each component alone. More than 75% of CAZ combinations exhibited additive interactions against biofilms of P. aeruginosa isolates, whereas CIP had mostly antagonistic interaction or no interaction with PMNs against biofilms of P. aeruginosa. Our findings demonstrate a greater positive interaction between AMK with PMNs than that observed for CAZ and especially CIP against isolates of P. aeruginosa from the respiratory tract of CF patients. PMID:25645829

  14. Human polymorphonuclear leukocytes inhibit Aspergillus fumigatus conidial growth by lactoferrin-mediated iron depletion.

    PubMed

    Zarember, Kol A; Sugui, Janyce A; Chang, Yun C; Kwon-Chung, Kyung J; Gallin, John I

    2007-05-15

    Aspergillus fumigatus, a common mold, rarely infects humans, except during prolonged neutropenia or in cases of chronic granulomatous disease (CGD), a primary immunodeficiency caused by mutations in the NADPH oxidase that normally produces fungicidal reactive oxygen species. Filamentous hyphae of Aspergillus are killed by normal, but not CGD polymorphonuclear leukocytes (PMN); however, the few studies on PMN-mediated host defenses against infectious conidia (spores) of this organism have yielded conflicting results, some showing that PMN do not inhibit conidial growth, with others showing that they do, most likely using reactive oxygen species. Given that CGD patients are exposed daily to hundreds of viable A. fumigatus conidia, yet considerable numbers of them survive years without infection, we reasoned that PMN use ROS-independent mechanisms to combat Aspergillus. We show that human PMN from both normal controls and CGD patients are equipotent at arresting the growth of Aspergillus conidia in vitro, indicating the presence of a reactive oxygen species-independent factor(s). Cell-free supernatants of degranulated normal and CGD neutrophils both suppressed fungal growth and were found to be rich in lactoferrin, an abundant PMN secondary granule protein. Purified iron-poor lactoferrin at concentrations occurring in PMN supernatants (and reported in human mucosal secretions in vivo) decreased fungal growth, whereas saturation of lactoferrin or PMN supernatants with iron, or testing in the presence of excess iron in the form of ferritin, completely abolished activity against conidia. These results demonstrate that PMN lactoferrin sequestration of iron is important for host defense against Aspergillus. PMID:17475866

  15. Electron microscopic identification of hydrogen peroxide detected in fixed human polymorphonuclear leukocytes during phagocytosis.

    PubMed

    Moriguchi, Keiichi; Ohno, Norikazu

    2014-01-01

    Polymorphonuclear leukocytes (PMNs) engaged in phagocytosis produce reactive oxygen species (ROS), such as those that occur in an activated NADPH oxidase reaction, to eliminate ingested microorganisms. The translocation of NADPH oxidase components to produce antimicrobial free radicals from the vesicles to the phagosomes may be important. Hydrogen peroxide (H2O2) derived from O2- has been observed by electron microscopy using a cerium method. However, 2'-7'-dichlorofluorescin diacetate can also detect H2O2 through fluorescence. The main objective of the present study was to measure the H2O2-dependent fluorescence of PMNs after opsonized zymosan A (OPZ) phagocytosis using a microplate reader under different fixation conditions, including 0.5, 1, and 10% glutaraldehyde (GA) individually for 1, 5, 10, or 30 min. An additional objective was to visualize, through the use of electron microscopic cytochemistry, the process of H2O2 generation in OPZ phagocytic fixed PMNs. The fixed PMNs showed that the largest fluorescent value was produced by a concentration of 0.5% GA for all fixation times. This suggested that the fixation of PMNs with a high concentration of GA inhibited phagocytosis and produced ROS. In the fixed PMNs, electron microscopic results showed that after 1 min of mixing, some PMNs attached to particles and exhibited mild deposits in their secretory vesicles. When PMNs engulfed particles, free radical-producing vesicles had enhanced reaction deposits 10 min later and fused to the phagosomal membrane, releasing numerous free radicals into the lumen. Time-dependent H2O2 production was enhanced in the secretory vesicles, some of which were fused exactly to the phagosome membranes. PMID:24815108

  16. Formylpeptide Receptor Single Nucleotide Polymorphism 348T>C and Its Relationship to Polymorphonuclear Leukocyte Chemotaxis in Aggressive Periodontitis

    PubMed Central

    Maney, Pooja; Walters, John D.

    2009-01-01

    Background Aggressive periodontitis (AgP) is associated with impaired polymorphonuclear leukocyte (PMN) chemotaxis toward bacterial N-formylpeptides. Formylpeptide receptors (FPRs) play a major role in guiding PMNs to infection sites. Previous work revealed a significant association between FPR1 single nucleotide polymorphism (SNP) 348T>C and AgP in African Americans. We tested the hypothesis that 348T impairs PMN chemotaxis by decreasing FPR mRNA expression, thereby increasing susceptibility to AgP. Methods Blood samples were obtained from African American subjects (37 AgP cases and 38 controls). Chemotaxis to N-formyl-Met-Leu-Phe by freshly isolated PMNs was assayed in a modified Boyden chamber. RNA was isolated from PMNs and FPR1 gene expression was quantified by real time PCR. To detect FPR1 5’ SNPs, genomic DNA was isolated and four fragments spanning the FPR1 5’ region were PCR-amplified and sequenced. Haplotype associations between SNP 348T>C and 5’ SNPs were analyzed. Results The homozygous 348T genotype was only found in AgP cases (P=0.017, odds ratio=18.9). Subjects with this genotype exhibited a significantly lower PMN chemotactic response relative to controls and to subjects with the 348C/C or 348T/C genotypes (P<0.05). There were no significant differences in PMN FPR1 expression between subjects with the 348C/C, 348C/T and 348 T/T genotypes. Eleven FPR1 5’ SNPs were detected, but none of the predicted haplotypes reflected associations with AgP or with 348T. Conclusions Although the 348T/T genotype is relatively rare, it is associated with significantly impaired PMN chemotaxis and an increased risk of developing AgP in African Americans. These associations do not appear to be related to significant reductions in FPR1 transcripts in subjects expressing 348T. PMID:19722801

  17. Effect of Antibiotics on Polymorphonuclear Leukocyte Functions and Myeloperoxidase Activity, Glutathione and Malondialdehyde Levels in Allergic Asthma.

    PubMed

    Rayaman, Pervin; Rayaman, Erkan; Cevikba?, Adile; Demirtunç, Refik; Sehirli, Ahmet Ozer; Alagöz, Seyda Gül; Gürer, Umran Soyo?ul

    2015-01-01

    We investigated the effect of ciprofloxacin, rifampicine and doxycycline on myeloperoxidase (MPO) activity, glutathione (GSH) and malondialdehyde (MDA) levels in allergic asthma patients and healthy volunteers. Polymorphonuclear leukocytes (PMNs) were isolated with ficoll-hypaque gradient centrifugation method. MPO activity was assayed with modified o-dianisidine, GSH by Ellman's and MDA levels by Beuge's method. PMN functions and MDA levels of patients significantly decreased when compared with healthy volunteers. Ciprofloxacin significantly increased PMN functions, MPO activity and MDA levels of both groups. We have demonstrated that ciprofloxacin has beneficial effects on MPO activity and PMN functions in allergic asthma patients and healthy volunteers. PMID:26094319

  18. Dendritic Cells Take up and Present Antigens from Viable and Apoptotic Polymorphonuclear Leukocytes

    PubMed Central

    Alfaro, Carlos; Suarez, Natalia; Ońate, Carmen; Perez-Gracia, Jose L.; Martinez-Forero, Ivan; Hervas-Stubbs, Sandra; Rodriguez, Inmaculada; Perez, Guiomar; Bolańos, Elixabet; Palazon, Asis; de Sanmamed, Miguel Fernandez; Morales-Kastresana, Aizea; Gonzalez, Alvaro; Melero, Ignacio

    2011-01-01

    Dendritic cells (DC) are endowed with the ability to cross-present antigens from other cell types to cognate T cells. DC are poised to meet polymorphonuclear leukocytes (PMNs) as a result of being co-attracted by interleukin-8 (IL-8), for instance as produced by tumor cells or infected tissue. Human monocyte-derived and mouse bone marrow-derived DC can readily internalize viable or UV-irradiated PMNs. Such internalization was abrogated at 4°C and partly inhibited by anti-CD18 mAb. In mice, DC which had internalized PMNs containing electroporated ovalbumin (OVA) protein, were able to cross-present the antigen to CD8 (OT-1) and CD4 (OT-2) TCR-transgenic T cells. Moreover, in humans, tumor cell debris is internalized by PMNs and the tumor-cell material can be subsequently taken up from the immunomagnetically re-isolated PMNs by DC. Importantly, if human neutrophils had endocytosed bacteria, they were able to trigger the maturation program of the DC. Moreover, when mouse PMNs with E. coli in their interior are co-injected in the foot pad with DC, many DC loaded with fluorescent material from the PMNs reach draining lymph nodes. Using CT26 (H-2d) mouse tumor cells, it was observed that if tumor cells are intracellularly loaded with OVA protein and UV-irradiated, they become phagocytic prey of H-2d PMNs. If such PMNs, that cannot present antigens to OT-1 T cells, are immunomagnetically re-isolated and phagocytosed by H-2b DC, such DC productively cross-present OVA antigen determinants to OT-1 T cells. Cross-presentation to adoptively transferred OT-1 lymphocytes at draining lymph nodes also take place when OVA-loaded PMNs (H-2d) are coinjected in the footpad of mice with autologous DC (H-2b). In summary, our results indicate that antigens phagocytosed by short-lived PMNs can be in turn internalized and productively cross-presented by DC. PMID:22206007

  19. Chemical, biochemical, pharmacokinetic, and biological properties of L-680,833: a potent, orally active monocyclic beta-lactam inhibitor of human polymorphonuclear leukocyte elastase.

    PubMed Central

    Doherty, J B; Shah, S K; Finke, P E; Dorn, C P; Hagmann, W K; Hale, J J; Kissinger, A L; Thompson, K R; Brause, K; Chandler, G O

    1993-01-01

    A series of potent and highly selective time-dependent monocyclic beta-lactam inhibitors of human polymorphonuclear leukocyte elastase (PMNE, EC 3.4.21.37) is described. The intrinsic potency of these compounds, as exemplified by L-680,833 (k(inactivation)/K(i) of 622,000 M-1.s-1), is reflected at the cellular level where it inhibits generation of the specific N-terminal cleavage product A alpha-(1-21) from the A alpha chain of fibrinogen by enzyme released from isolated polymorphonuclear leukocytes stimulated with fMet-Leu-Phe with an IC50 of 0.06 microM. The inhibitory activity of L-680,833 is also apparent in whole blood stimulated with A23187, where it inhibits formation of A alpha-(1-21) and PMNE-alpha 1-proteinase inhibitor complex formation with IC50 values of 9 microM. Pharmacokinetic studies indicate that after oral dosing L-680,833 is bioavailable in rats and rhesus monkeys. This oral bioavailability is reflected by the inhibition (i) of tissue damage elicited in hamster lungs by intratracheal instillation of human PMNE and (ii) enzyme released from human PMN stimulated after their transfer into the pleural cavity of mice. The properties of L-680,833 allow it to effectively supplement the activity of natural inhibitors of PMNE in vivo, suggesting that this type of low-molecular-weight synthetic inhibitor could have therapeutic value in diseases where PMNE damages tissue. PMID:8378355

  20. [Interaction of polymorphonuclear leukocytes, bradykinin and carragenin. Transference reaction in the rat].

    PubMed

    Lismonde, M; Damas, J; Remacle-Volon, G

    1982-10-01

    1. Injected in the paw of the rat, polymorphonuclear leucocytes do not increase the oedematogen action of bradykinin, but increase the action of lambda carrageenan. 2. This potentiation of carrageenan action is not modified when PG biosynthesis in leucocytes is inhibited by indomethacin or aspirin. It does not appear in rats previously treated by indomethacin or aspirin. 3. Our results suggest that rat polymorphonuclear leucocytes increase the inflammatory reaction, when they are stimulated by carrageenan, by the release of a phospholipase A2 activity which induces PG biosynthesis in rat paw tissues. PMID:6186210

  1. Promotion of DNA strand breaks in cocultured mononuclear leukocytes by protein kinase C-dependent prooxidative interactions of benoxaprofen, human polymorphonuclear leukocytes, and ultraviolet radiation

    SciTech Connect

    Schwalb, G.; Beyers, A.D.; Anderson, R.; Nel, A.E.

    1988-06-01

    At concentrations of 5 micrograms/ml and greater the nonsteroidal antiinflammatory drug benoxaprofen caused dose-related activation of lucigenin-enhanced chemiluminescence in human polymorphonuclear leukocytes (PMNL). Benoxaprofen-mediated activation of lucigenin-enhanced chemiluminescence by PMNL was increased by UV radiation and was particularly sensitive to inhibition by the selective protein kinase C inhibitor H-7. To identify the molecular mechanism of the prooxidative activity of benoxaprofen, the effects of the nonsteroidal antiinflammatory drug on the activity of purified protein kinase C in a cell-free system were investigated. Benoxaprofen caused a dose-related activation of protein kinase C by interaction with the binding site for the physiological activator phosphatidylserine, but could not replace diacylglycerol. When autologous mononuclear leukocytes (MNL) were cocultured with PMNL and benoxaprofen in combination, but not individually, the frequency of DNA strand breaks in MNL was markedly increased. UV radiation significantly potentiated damage to DNA mediated by benoxaprofen and PMNL. Inclusion of superoxide dismutase, H-7, and, to a much lesser extent, catalase during exposure of MNL to benoxaprofen-activated PMNL prevented oxidant damage to DNA. These results clearly demonstrate that potentially carcinogenic prooxidative interactions, which are unlikely to be detected by conventional assays of mutagenicity, may occur between phagocytes, UV radiation, and certain pharmacological agents.

  2. Possible mechanisms for the differential effects of high linoleate safflower oil and high ?-linolenate perilla oil diets on platelet-activating factor production by rat polymorphonuclear leukocytes

    Microsoft Academic Search

    Kentaro Oh-hashi; Tetsuya Takahashi; Shiro Watanabe; Tetsuyuki Kobayashi; Harumi Okuyama

    1997-01-01

    As compared with high dietary linoleate safflower oil, high dietary ?-linolenate perilla oil decreased platelet-activating factor (PAF) production by nearly half in calcium ionophore (CaI)-stimulated rat polymorphonuclear leukocytes (PMN). In the CaI-stimulated PMN from the perilla oil group, the accumulated amount of arachidonate (AA) plus eicosapentaenoate (EPA) was 30% less and that of lyso-PAF was 50% less, indicating that the

  3. Inactivation of the rhlA gene in Pseudomonas aeruginosa prevents rhamnolipid production, disabling the protection against polymorphonuclear leukocytes

    PubMed Central

    VAN GENNIP, MARIA; CHRISTENSEN, LOUISE DAHL; ALHEDE, MORTEN; PHIPPS, RICHARD; JENSEN, PETER ŘSTRUP; CHRISTOPHERSEN, LARS; PAMP, SÜNJE JOHANNA; MOSER, CLAUS; MIKKELSEN, PER JENSEN; KOH, ANDREW Y.; TOLKER-NIELSEN, TIM; PIER, GERALD B.; HŘIBY, NIELS; GIVSKOV, MICHAEL; BJARNSHOLT, THOMAS

    2010-01-01

    Many of the virulence factors produced by the opportunistic human pathogen Pseudomonas aeruginosa are quorum-sensing (QS) regulated. Among these are rhamnolipids, which have been shown to cause lysis of several cellular components of the human immune system, e.g. monocyte-derived macrophages and polymorphonuclear leukocytes (PMNs). We have previously shown that rhamnolipids produced by P. aeruginosa cause necrotic death of PMNs in vitro. This raises the possibility that rhamnolipids may function as a ‘biofilm shield’ in vivo, which contributes significantly to the increased tolerance of P. aeruginosa biofilms to PMNs. In the present study, we demonstrate the importance of the production of rhamnolipids in the establishment and persistence of P. aeruginosa infections, using an in vitro biofilm system, an intraperitoneal foreign-body model and a pulmonary model of P. aeruginosa infections in mice. Our experimental data showed that a P. aeruginosa strain unable to produce any detectable rhamnolipids, due to an inactivating mutation in the single QS-controlled rhlA gene, did not induce necrosis of PMNs in vitro and exhibited increased clearance compared with its wild-type counterpart in vivo. Conclusively, the results support our model that rhamnolipids are key protective agents of P. aeruginosa against PMNs. PMID:19594494

  4. beta. -Endorphin and related peptides suppress phorbol myristate acetate-induced respiratory burst in human polymorphonuclear leukocytes

    SciTech Connect

    Diamant, M.; Henricks, P.A.J.; Nijkamp, F.P.; de Wied, D. (Univ. of Utrecht (Netherlands))

    1989-01-01

    In the present study, the immunomodulatory effect of {beta}-endorphin ({beta}-E) and shorter pro-opiomelancortin (POMC) fragments was evaluated by assessing their influence on respiratory burst in human polymorphonuclear leukocytes (PMN). The effect of the peptides on phorbol myristate acetate (PMA)-stimulated production of reactive oxygen metabolites was measured in a lucigenin-enhanced chemiluminescence (CL) assay. Both POMC peptides with opiate-like activity and their non-opioid derivatives were tested. With the exception of {alpha}-E, PMA-stimulated respiratory burst was suppressed by all POMC fragments tested. A U-shaped dose-response relation was observed. Doses lower than 10{sup {minus}17}M and higher than 10{sup {minus}8}M were without effect. {beta}-E and dT{beta}E both suppressed PMA-induced oxidative burst in human PMN at physiological concentrations. {gamma}-E and dT{gamma}E proved to be less potent inhibitors, reaching maximal effect at higher concentrations. DE{gamma}E exerted an even less pronounced but still significant suppressive effect at the concentration of 10{sup {minus}10}M. None of the endorphins tested was shown to affect resting oxidative metabolism in the PMN. The modulatory effects of the opioid peptides could not be blocked by the opioid antagonist naloxone.

  5. Harvesting the noncirculating pool of polymorphonuclear leukocytes in rats by hetastarch exchange transfusion (HET): yield and functional assessment

    SciTech Connect

    Williams, J.H. Jr.; Moser, K.M.; Ulich, T.; Cairo, M.S.

    1987-11-01

    Isolation of polymorphonuclear leukocytes (PMN) provides an opportunity to study PMN activity in vitro and to label PMN for study of in vivo kinetics. However, simple phlebotomy (SP) of a small animal frequently yields too few PMN for in vitro handling, while PMN harvested from an induced-peritonitis may not accurately reflect PMN in a less stimulated state. We report a novel method of harvesting PMN from the circulation of rats, using hetastarch exchange transfusion (HET), which is both time and animal sparing. HET harvested 8-fold more PMN than SP. In vitro cell function was examined with assays of adherence, chemotaxis, bacterial killing, and superoxide generation. No significant (p less than 0.05) difference was found between PMN obtained by HET and pooled-PMN obtained by SP. In vivo function was examined following labeling with indium 111-oxine. The kinetics pattern described suggested normal migratory activity when compared to previous reports. The data demonstrate that rats possess a relatively large, noncirculating pool of PMN which is readily accessible by HET.

  6. Deep venous thrombosis: behaviour of the polymorphonuclear leukocyte integrin pattern at baseline and after in vitro activation.

    PubMed

    Caimi, G; Tozzi Ciancarelli, M G; Ferrara, F; Montana, M; Calandrino, V; Canino, B; Lo Presti, R

    2005-01-01

    In a group of 18 subjects with acute deep venous thrombosis (DVT), evidenced by clinical examination and echo-color-Doppler, we examined the phenotypical expression of the polymorphonuclear leukocyte (PMN) beta2-integrins (CD11a, CD11b, CD11c, CD18), obtained by using a flow cytofluorimeter. The evaluation was performed before and after in vitro activation (prolonged for 5 and 15 minutes) with 4-phorbol 12-myristate 13-acetate (PMA) and N-formyl-methionyl-leucyl-phenylalanine (fMLP). In DVT subjects, at baseline, the phenotypical expression of CD11b was decreased and that of CD11c was increased when compared with normal controls; no difference was found in CD11a and CD18 expression. In normal subjects PMN activation with both activators led to a constant increase of all PMN adhesion molecules; in DVT subjects CD11b, CD11c and CD18 increased, while CD11a expression did not show any change. These data indicate the presence of a functional alteration in circulating PMN cells from patients with DVT. PMID:16037628

  7. Genome-wide protective response used by group A Streptococcus to evade destruction by human polymorphonuclear leukocytes.

    PubMed

    Voyich, Jovanka M; Sturdevant, Daniel E; Braughton, Kevin R; Kobayashi, Scott D; Lei, Benfang; Virtaneva, Kimmo; Dorward, David W; Musser, James M; DeLeo, Frank R

    2003-02-18

    Group A Streptococcus (GAS) evades polymorphonuclear leukocyte (PMN) phagocytosis and killing to cause human disease, including pharyngitis and necrotizing fasciitis (flesh-eating syndrome). We show that GAS genes differentially regulated during phagocytic interaction with human PMNs comprise a global pathogen-protective response to innate immunity. GAS prophage genes and genes involved in virulence, oxidative stress, cell wall biosynthesis, and gene regulation were up-regulated during PMN phagocytosis. Genes encoding novel secreted proteins were up-regulated, and the proteins were produced during human GAS infections. We discovered an essential role for the Ihk-Irr two-component regulatory system in evading PMN-mediated killing and promoting host-cell lysis, processes that would facilitate GAS pathogenesis. Importantly, the irr gene was highly expressed during human GAS pharyngitis. We conclude that a complex pathogen genetic program circumvents human innate immunity to promote disease. The gene regulatory program revealed by our studies identifies previously undescribed potential vaccine antigens and targets for therapeutic interventions designed to control GAS infections. PMID:12574517

  8. Polymorphonuclear leukocyte membrane fluidity and cytosolic Ca2+ concentration in diabetes mellitus.

    PubMed

    Caimi, G; Canino, B; Montana, M; Ventimiglia, G; Catania, A; Lo Presti, R

    1998-10-01

    We evaluated polymorphonuclear membrane (PMN) fluidity in 32 subjects with type 1 diabetes mellitus, 38 subjects with type 2 diabetes mellitus and 38 normal control subjects, by marking intact and unstimulated PMN cells with the fluorescent probe 1-[4-(trimethylamino)phenyl]-6-phenyl-1,3,5-hexatriene (TMA-DPH). We also evaluated PMN cytosolic Ca2+ content by marking intact and unstimulated PMN cells with the fluorescent probe Fura 2-AM. PMN membrane fluidity differentiated normal subjects from type 1 and 2 diabetic subjects. The PMN cytosolic Ca2+ concentration did not discriminate type 1 and 2 diabetic subjects from normal control subjects. No statistical correlation was found between PMN membrane fluidity and PMN cytosolic Ca2+ concentration in any of the groups of subjects, nor were significant correlations found between PMN membrane fluidity and cytosolic Ca2+ concentration in several plasma parameters (serum glucose, cholesterol and triglycerides). In conclusion, in type 1 and 2 diabetic patients we found a decrease in PMN membrane fluidity and this decrease, which was greater in type 2 diabetic patients, may be a marker of PMN dysfunction. PMID:9840453

  9. VARIATION AMONG GOATS IN THE ABILITY OF THEIR POLYMORPHONUCLEAR NEUTROPHIL LEUKOCYTES AND MAMMARY SECRETIONS TO SUPPORT PHAGOCYTOSIS: INHIBITORY EFFECTS OF MILK FAT GLOBULES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objectives of this study were to determine if fat globules and casein in goat milk were inhibitory to phagocytosis by polymorphonuclear neutrophils (PMN) isolated from blood, and to determine if variation existed among goats in the ability of PMN to phagocytose and in the ability of milk whey ...

  10. 77 FR 59000 - Guidance for Industry: Pre-Storage Leukocyte Reduction of Whole Blood and Blood Components...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-25

    ...Industry: Pre-Storage Leukocyte Reduction of Whole Blood and Blood Components Intended for Transfusion; Availability AGENCY...Industry: Pre- Storage Leukocyte Reduction of Whole Blood and Blood Components Intended for Transfusion''...

  11. 76 FR 5386 - Draft Guidance for Industry: Pre-Storage Leukocyte Reduction of Whole Blood and Blood Components...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-31

    ...Industry: Pre-Storage Leukocyte Reduction of Whole Blood and Blood Components Intended for Transfusion; Availability AGENCY...Industry: Pre- Storage Leukocyte Reduction of Whole Blood and Blood Components Intended for Transfusion''...

  12. Mechanism for the inflammatory response in primate lungs. Demonstration and partial characterization of an alveolar macrophage-derived chemotactic factor with preferential activity for polymorphonuclear leukocytes.

    PubMed Central

    Kazmierowski, J A; Gallin, J I; Reynolds, H Y

    1977-01-01

    Approximately 4 h after an initial bronchoalveolar lavage (BAL) of a primate's lung, an appreciable number of polymorphonuclear leukocytes (PMNs) were noted to accumulate in respiratory fluids when lavage was repeated. Whereas, alveolar macrophages (90%) and lymphocytes (7%) were the principal respiratory cells recovered initially from lavage fluid, later samples contained 45-90% PMNs To explain the observed ingress of PMNs into lung fluids, concentrated BAL fluid was tested for chemoattractant activity. Such fluid obtained 4 and 24 h after an initial lavage contained material that produced directed migration (chemotaxis) for PMNs and mononuclear cells isolated from peripheral blood of normal donors. Gel filtration chromatography of BAL disclosed two peaks of chemotactic activity in the effluent fractions. Material from the column with an estimated molecular weight of 15,000 daltons was chemotactic for both PMNs and mononuclear cells. Because it was susceptible to inactivation with antiserum against the fifth component of complement, resistant to heating, and unaffected by antiserum against C3, this factor was considered analogous to the cleavage product of the fifth component of complement. C5a. In addition chemotactic activity for PMNs only was contained in an effluent peak having a molecular weight of about 5,000 daltons. This material was heat labile but unaffected by antisera to complement components. To locate the possible source of these factors in respiratory fluid, in vitro cultures of alveolar macrophages were established. These cells, whether stimulated by phagocytosis of opsonized bacteria or merely by attachment to a glass surface, produced chemotactic material which had physical characteristics similar to the small molecular weight material in BAL. Moreover, it induced preferential chemotaxis for PMNs. Thus, in primate lungs, at least two chemotactic substances may generate an inflammatory response; one which is a fragment of the complement component C5 and another small molecular weight factor which is released from alveolar macrophages. PMID:401834

  13. A New Form of Localized Allergic Encephalomyelitis Featuring Polymorphonuclear Neutrophilic Leukocytes

    PubMed Central

    Levine, Seymour; Hoenig, Eugene M.

    1971-01-01

    Mononuclear inflammatory infiltrates are characteristic of experimental allergic encephalomyelitis (EAE). Localized EAE was produced in rats in a single day by passive transfer of living lymphoid cells from immunized donors to non-immunized recipients with thermal brain injuries. When the recipients were pretreated with cyclophosphamide 1 or 2 days before cell transfer, neutrophils in the EAE infiltrates notably increased and mononuclear leukocytes decreased. This neutrophilic form of EAE was produced with cells from donors immunized with whole neural tissue or purified myelin basic protein and with adjuvants of different types. This lesion could not be reproduced if the EAE cells were replaced by EAE serum or by irrelevantly immunized cells, or if their activity was foiled by specific “desensitization” or by use of histoincompatible recipients. The 2-day period during which cyclophosphamide prepared recipients for neutrophilic EAE coincided with a transient lymphopenia and relative neutrocytosis caused by the drug. The passive transfer system made it possible for the drug to unbalance the recipient's hemopoietic system without risking adverse effects on the donor cells. The rapid development of the localized form of EAE made it possible to produce lesions before the recipient's transient imbalance gave way to pancytopenia. The new form of EAE may be useful for investigating autoimmune diseases because the neutrophils which indicate the immunologic injury are instantly recognized as reactive recipient cells. Preexisting conventional (mononuclear) EAE prevented subsequent production of neutrophilic infiltrates. This inhibitory effect may be due to local blockade of vessels by mononuclear cells, a concept for which there is evidence. These considerations suggest that the exudation of neutrophils in the new form of EAE may be due to suppression by cyclophosphamide of the mononuclear component of the inflammatory reaction, with loss of its blockading and protective influence on the vessels. ImagesFig 5Fig 6Fig 1Fig 2Fig 3Fig 4 PMID:5556631

  14. Iron Sucrose Impairs Phagocytic Function and Promotes Apoptosis in Polymorphonuclear Leukocytes

    PubMed Central

    Ichii, Hirohito; Masuda, Yuichi; Hassanzadeh, Tania; Saffarian, Mateen; Gollapudi, Sastry; Vaziri, Nosratola D

    2014-01-01

    Background With the recent implementation of bundling reimbursement policy the use of intravenous (IV) iron preparations for the management of anemia in the ESRD population has dramatically increased. Iron overload increases the risk of infections in individuals with or without kidney disease. IV iron administration in ESRD patients impairs bacteriocidal capacity of PMNs against Escherichia Coli. These preparations consist of an elemental iron core and a carbohydrate shell. In addition to the iron core the carbohydrate shell may affect PMNs. We therefore examined the effect of iron sucrose, a commonly used preparation, on phagocytic capacity of PMNs from a group of normal individuals against Gram positive (Staphylococcus Aureus) and Gram negative (E. Coli) bacteria. Methods Iron sucrose was added to heparinized blood samples at pharmacologically-relevant concentrations and incubated for 4 and 24 hours at 37° C to simulate in vivo condition. Blood samples mixed with equal volume of saline solution served as controls. To isolate the effects of the carbohydrate shell, blood samples were co-treated with the iron chelator, desferrioxamine. Results Iron sucrose caused significant PMN apoptosis and dose-dependent suppression of phagocytic function against both Gram positive and negative bacteria. These abnormalities were prevented by desferrioxamine which precluded contribution of the carbohydrate shell to the PMN dysfunction. Conclusions At pharmacologically-relevant concentrations iron sucrose promotes apoptosis and inhibits phagocytic activities of PMNs. The deleterious effect of iron sucrose is mediated by its elemental iron core, not its carbohydrate shell, and as such may be shared by other IV iron preparations. PMID:22722756

  15. Leukotriene B/sub 4/ modulates human polymorphonuclear leukocyte (PMN) phospholipid (PL) methylation

    SciTech Connect

    Bomalaski, J.S.; Clark, M.A.; Dundee, D.

    1986-05-01

    Formation of phosphatidylcholine (PC) by methylation of phosphatidylethanolamine (PE) is required for transduction of chemotactic factor messages in phagocytic cells, including the development of a motile configuration and release of arachidonic acid (AA) from PL. The authors examined PL methylation in human peripheral blood PMN following stimulation by leukotriene B/sub 4/ (LTB/sub 4/), a potent chemotactic lipid metabolite of AA that is produced by human PMN. (/sup 3/H)-methionine, in the presence or absence of LTB/sub 4/, was added directly to the cells without a preloading period. (/sup 3/H)-methyl mono- and di-methylated PE, PC and lyso PC were separated by thin layer chromatography. PL methylation in human PMN is dependent on time of incubation, LTB/sub 4/ concentration and methionine concentration. The optimal LTB/sub 4/ concentration is 10/sup -7/ M, the same concentration that induces a maximal chemotactic response in PMN. At early time points (2-10 min), formation of methylated PL is enhanced following LTB/sub 4/ stimulation. In contrast, at later time points (20-60 min), methylated PL synthesis in LTB/sub 4/ treated cells is depressed or the same as control cells. Synthesis of PC by the choline transferase pathway is not affected by LTB/sub 4/ stimulation. This data shows that, at early time points following LTB/sub 4/ stimulation, PL methylation is enhanced. This correlates with other reports of calcium mobilization and chemotaxis in PMN at early time points following LTB/sub 4/ stimulation.

  16. Human cytomegalovirus replicates abortively in polymorphonuclear leukocytes after transfer from infected endothelial cells via transient microfusion events.

    PubMed

    Gerna, G; Percivalle, E; Baldanti, F; Sozzani, S; Lanzarini, P; Genini, E; Lilleri, D; Revello, M G

    2000-06-01

    Using a recently developed model for in vitro generation of pp65-positive polymorphonuclear leukocytes (PMNLs), we demonstrated that PMNLs from immunocompetent subjects may harbor both infectious human cytomegalovirus (HCMV) and viral products (pp65, p72, DNA, and immediate-early [IE] and pp67 late mRNAs) as early as 60 min after coculture with human umbilical vein endothelial cells (HUVEC) or human embryonic lung fibroblasts (HELF) infected with a clinical HCMV isolate (VR6110) or other wild-type strains. The number of PMNLs positive for each viral parameter increased with coculture time. Using HELF infected with laboratory-adapted HCMV strains, only very small amounts of viral DNA and IE and late mRNAs were detected in PMNLs. A cellular mRNA, the vascular cell adhesion molecule-1 mRNA, which is abundantly present in both infected and uninfected HUVEC, was detected in much larger amounts in PMNLs cocultured with VR6110-infected cells than in controls. Coculture of PMNLs with VR6110-infected permissive cells in the presence or absence of RNA, protein, and viral DNA synthesis inhibitors showed that only IE genes were transcribed in PMNLs during coculture. Synthesis of IE transcripts in PMNLs was also supported by the finding that only the copy number of IE mRNA (and not the DNA or the pp67 mRNA) per infected PMNL increased markedly with time, and the pp67 to IE mRNA copy number ratio changed from greater than 10 in infected HUVEC to less than 1 in cocultured PMNLs. Fluorescent probe transfer experiments and electron microscopy studies indicated that transfer of infectious virus and viral products from infected cells to PMNLs is likely to be mediated by microfusion events induced by wild-type strains only. In addition, HCMV pp65 and p72 were both shown to localize in the nucleus of the same PMNLs by double immunostaining. Two different mechanisms may explain the virus presence in PMNLs: (i) one major mechanism consists of transitory microfusion events (induced by wild-type strains only) of HUVEC or HELF and PMNLs with transfer of viable virus and biologically active viral material to PMNLs; and (ii) one minor mechanism, i.e., endocytosis, occurs with both wild-type and laboratory strains and leads to the acquisition of very small amounts of viral nucleic acids. In conclusion, HCMV replicates abortively in PMNLs, and wild-type strains and their products (as well as cellular metabolites and fluorescent dyes) are transferred to PMNLs, thus providing evidence for a potential mechanism of HCMV dissemination in vivo. PMID:10823870

  17. Human Cytomegalovirus Replicates Abortively in Polymorphonuclear Leukocytes after Transfer from Infected Endothelial Cells via Transient Microfusion Events

    PubMed Central

    Gerna, Giuseppe; Percivalle, Elena; Baldanti, Fausto; Sozzani, Silvano; Lanzarini, Paolo; Genini, Emilia; Lilleri, Daniele; Revello, Maria Grazia

    2000-01-01

    Using a recently developed model for in vitro generation of pp65-positive polymorphonuclear leukocytes (PMNLs), we demonstrated that PMNLs from immunocompetent subjects may harbor both infectious human cytomegalovirus (HCMV) and viral products (pp65, p72, DNA, and immediate-early [IE] and pp67 late mRNAs) as early as 60 min after coculture with human umbilical vein endothelial cells (HUVEC) or human embryonic lung fibroblasts (HELF) infected with a clinical HCMV isolate (VR6110) or other wild-type strains. The number of PMNLs positive for each viral parameter increased with coculture time. Using HELF infected with laboratory-adapted HCMV strains, only very small amounts of viral DNA and IE and late mRNAs were detected in PMNLs. A cellular mRNA, the vascular cell adhesion molecule-1 mRNA, which is abundantly present in both infected and uninfected HUVEC, was detected in much larger amounts in PMNLs cocultured with VR6110-infected cells than in controls. Coculture of PMNLs with VR6110-infected permissive cells in the presence or absence of RNA, protein, and viral DNA synthesis inhibitors showed that only IE genes were transcribed in PMNLs during coculture. Synthesis of IE transcripts in PMNLs was also supported by the finding that only the copy number of IE mRNA (and not the DNA or the pp67 mRNA) per infected PMNL increased markedly with time, and the pp67 to IE mRNA copy number ratio changed from greater than 10 in infected HUVEC to less than 1 in cocultured PMNLs. Fluorescent probe transfer experiments and electron microscopy studies indicated that transfer of infectious virus and viral products from infected cells to PMNLs is likely to be mediated by microfusion events induced by wild-type strains only. In addition, HCMV pp65 and p72 were both shown to localize in the nucleus of the same PMNLs by double immunostaining. Two different mechanisms may explain the virus presence in PMNLs: (i) one major mechanism consists of transitory microfusion events (induced by wild-type strains only) of HUVEC or HELF and PMNLs with transfer of viable virus and biologically active viral material to PMNLs; and (ii) one minor mechanism, i.e., endocytosis, occurs with both wild-type and laboratory strains and leads to the acquisition of very small amounts of viral nucleic acids. In conclusion, HCMV replicates abortively in PMNLs, and wild-type strains and their products (as well as cellular metabolites and fluorescent dyes) are transferred to PMNLs, thus providing evidence for a potential mechanism of HCMV dissemination in vivo. PMID:10823870

  18. Persistence of the altered polymorphonuclear leukocyte rheological and metabolic variables after 12 months in juvenile myocardial infarction.

    PubMed

    Lo Presti, Rosalia; Tozzi Ciancarelli, Maria Giuliana; Hoffmann, Enrico; Incalcaterra, Egle; Canino, Baldassare; Montana, Maria; D'Amico, Teresa; Catania, Anna; Caimi, Gregorio

    2006-01-01

    Acute myocardial infarction (AMI) is associated with an elevated polymorphonuclear leukocyte (PMN) count and a PMN rheological impairment. In this study we evaluated two major rheological aspects (membrane fluidity and cytosolic Ca2+ concentration) in a group of young adults with AMI. We enrolled 41 AMI patients (39 men and 2 women; mean age 41.0 +/- 4.0 years), who were examined 5-10 days after AMI (T1) and 12 months later (T2). The membrane fluidity was obtained labelling granulocytes with the fluorescent probe 1-[4-(trimethylamino)phenyl]-6-phenyl-1,3,5-hexatriene (TMA-DPH) and considering the degree of fluorescence polarization, inversely correlated to the membrane lipid fluidity. The cytosolic Ca2+ content was obtained marking PMN cells with the fluorescent probe Fura-2AM and considering the ratio between the Fura 2-Ca2+ complex and the unchelated Fura 2 fluorescence intensity. Both parameters were evaluated at baseline and after in vitro activation with 4-phorbol 12-myristate 13-acetate (PMA) at the concentration of 4.5 muM, prolonged for 5 and 15 minutes. At T1 the PMN membrane fluidity and cytosolic Ca2+ content in AMI patients were respectively decreased and increased in comparison with control group. At T2 the membrane fluidity was not any more different from control subjects, but there was also a further increase in cytosolic Ca2+ content. In vitro, PMN activation caused no significant variation of these parameters in the control group, while in AMI patients membrane fluidity significantly decreased and cytosolic Ca2+ content increased not only during the initial stage, but also after 12 months. The long-term functional alteration of PMN cells observed in young adults with AMI confirms the role of these cells in the inflammatory response following AMI. In the light of these data, the use of molecules able to modulate granulocyte activity, such as calcium channel blockers or pentoxifylline, should be reconsidered in myocardial infarction, together with the usual pharmacological treatment. PMID:16899933

  19. Effects of insulin on platelet and leukocyte activity in whole blood

    Microsoft Academic Search

    Hu Hu; Paul Hjemdahl; Nailin Li

    2002-01-01

    Diabetes mellitus (DM) is associated with platelet and leukocyte dysfunction. Previous observations with regard to insulin effects on platelet and leukocyte function are less than consistent. We thus investigated the effects of insulin on platelets and leukocytes, as well as on platelet–leukocyte interactions in whole blood. Hirudinized whole blood from 20 healthy subjects was preincubated at 37 °C in the

  20. Leukocyte transport by red blood cells in a microvessel

    NASA Astrophysics Data System (ADS)

    Freund, Jonathan

    2009-11-01

    A simulation model is used to study the transport of relatively large, spherical, and stiff white blood cells (leukocytes) by the relatively smaller and highly flexible red cell as they flow in the microcirculation. Their interaction dynamics are thought to be an important component of the inflammation response, in which leukocytes bind to the walls of blood vessels. The red cells are modeled in the simulations as highly deformable three-dimensional shells encasing a Newtonian fluid, and the viscous-flow equation is solved via a boundary integral formulation in which the cell shapes discretized by global spectral basis functions. For slow flow rates, it is found that the leukocyte is predominantly adjacent the vessel walls, whereas for faster flow rates this configuration appears to become unstable and the leukocyte traverses the whole vessel in a seemingly random fashion. For the straight round tubes simulated thus far, the stable leukocyte stand-off distance is always beyond the range of the binding molecules that capture it, which suggests that vessel inhomogeneities or interactions with other white cells are needed to create contact and thereby binding with the vessel walls.

  1. Oxidant-dependent metabolic activation of polycyclic aromatic hydrocarbons by phorbol ester-stimulated human polymorphonuclear leukocytes: possible link between inflammation and cancer

    SciTech Connect

    Trush, M.A.; Seed, J.L.; Kensler, T.W.

    1985-08-01

    Oxidants, such as those generated by metabolically activated phagocytes in inflammation, have been implicated in the metabolic activation of carcinogens, and in this study the authors demonstrate that the interaction of (+/-)-trans-7,8-dihydroxy-7,8-dihydrobenzo(a)pyrene (BP 7,8-dihydrodiol) with phorbol ester-stimulated polymorphonuclear leukocytes (PMNs) results in the generation of both a chemiluminescent intermediate and one that covalently binds to DNA. Concordant with the formation of a carcinogen-DNA adduct, the admixture of BP 7,8-dihydrodiol and phorbol ester-stimulated PMNs elicited mutagenesis in Salmonella typhimurium strain TA100. These results demonstrate that oxidants generated by metabolically stimulated PMNs can activate penultimate polycyclic aromatic hydrocarbons to a genotoxic metabolite and further defines a role for inflammation in carcinogenesis.

  2. Population Pharmacokinetics of Azithromycin in Whole Blood, Peripheral Blood Mononuclear Cells, and Polymorphonuclear Cells in Healthy Adults

    PubMed Central

    Sampson, M R; Dumitrescu, T P; Brouwer, K L R; Schmith, V D

    2014-01-01

    Azithromycin's extensive distribution to proinflammatory cells, including peripheral blood mononuclear cells (PBMCs) and polymorphonuclear cells (PMNs), may be important to its antimicrobial and anti-inflammatory properties. The need to simultaneously predict azithromycin concentrations in whole blood (“blood”), PBMCs, and PMNs motivated this investigation. A single-dose study in 20 healthy adults was conducted, and nonlinear mixed effects modeling was used to simultaneously describe azithromycin concentrations in blood, PBMCs, and PMNs (simultaneous PK model). Data were well described by a four-compartment mamillary model. Apparent central clearance and volume of distribution estimates were 67.3 l/hour and 336 l (interindividual variability of 114 and 122%, respectively). Bootstrapping and visual predictive checks showed adequate model performance. Azithromycin concentrations in blood, PBMCs, and PMNs from external studies of healthy adults and cystic fibrosis patients were within the 5th and 95th percentiles of model simulations. This novel empirical model can be used to predict azithromycin concentrations in blood, PBMCs, and PMNs with different dosing regimens. PMID:24599342

  3. Polymorphonuclear leukocyte membrane fluidity and cytosolic Ca(2+) content in young adults with acute myocardial infarction. Evaluation at the initial stage and after 12 months.

    PubMed

    Caimi, G; Hoffmann, E; Canino, B; Montana, M; Dispensa, F; Incalcaterra, E; Casciolo, M F; Catania, A; Lo Presti, R

    2004-01-01

    Our aim was to examine two aspects of polymorphonuclear leukocyte (PMN) rheology (membrane fluidity and cytosolic Ca2+ content), at baseline and after in vitro activation, in a group of young adults with acute myocardial infarction (AMI) at the initial stage and after 12 months. We enrolled 21 AMI subjects aged < or = 45 years (mean age 41.1 +/- 3.5 years) and evaluated PMN membrane fluidity, labelling intact PMN cells with the fluorescent probe 1,4-(trimethylamino)-phenyl-4-phenylhexatriene and the PMN cytosolic Ca2+ content marking PMN cells with the fluorescent probe Fura 2-AM, at baseline and after in vitro activation with 4-phorbol 12-myristate 13-acetate (PMA) and N-formyl-methionyl-leucyl-phenylalanine (fMLP). During the initial stage PMN membrane fluidity and cytosolic Ca2+ content did not distinguish AMI patients from control subjects; after 12 months, when compared with the initial stage, PMN cytosolic Ca2+ content was significantly increased. In vitro PMN activation with PMA and fMLP caused no variation of the two PMN parameters in control subjects, while in AMI patients membrane fluidity decreased and cytosolic Ca2+ content increased; the same behaviour pattern was observed after 12 months. The constant functional alteration of PMN cells in young AMI patients highlights the role of activated leukocytes as a component of the inflammatory reaction that follows ischemia. PMID:15272152

  4. Tracking flow of leukocytes in blood for drug analysis

    NASA Astrophysics Data System (ADS)

    Basharat, Arslan; Turner, Wesley; Stephens, Gillian; Badillo, Benjamin; Lumpkin, Rick; Andre, Patrick; Perera, Amitha

    2011-03-01

    Modern microscopy techniques allow imaging of circulating blood components under vascular flow conditions. The resulting video sequences provide unique insights into the behavior of blood cells within the vasculature and can be used as a method to monitor and quantitate the recruitment of inflammatory cells at sites of vascular injury/ inflammation and potentially serve as a pharmacodynamic biomarker, helping screen new therapies and individualize dose and combinations of drugs. However, manual analysis of these video sequences is intractable, requiring hours per 400 second video clip. In this paper, we present an automated technique to analyze the behavior and recruitment of human leukocytes in whole blood under physiological conditions of shear through a simple multi-channel fluorescence microscope in real-time. This technique detects and tracks the recruitment of leukocytes to a bioactive surface coated on a flow chamber. Rolling cells (cells which partially bind to the bioactive matrix) are detected counted, and have their velocity measured and graphed. The challenges here include: high cell density, appearance similarity, and low (1Hz) frame rate. Our approach performs frame differencing based motion segmentation, track initialization and online tracking of individual leukocytes.

  5. Myristic Acid, A Side Chain of Phorbol Myristate Acetate (PMA), Can Activate Human Polymorphonuclear Leukocytes to Produce Oxygen Radicals More Potently than PMA

    PubMed Central

    Tada, Mika; Ichiishi, Eiichiro; Saito, Rumiko; Emoto, Natsumi; Niwano, Yoshimi; Kohno, Masahiro

    2009-01-01

    Myristic acid (MyA), which is a saturated fatty acid (C14:0) and a side chain of phorbol 12-myristate 13-acetate (PMA), was examined if MyA stimulates human polymorphonuclear leukocytes (PMNs) to release oxygen radicals comparable to PMA by applying electron paramagnetic resonance (EPR)-spin-trapping method. When MyA was added to isolated human PMNs, spin adducts of 5,5-dimethyl-1-pyrroline-N-oxide (DMPO)-OH and DMPO-OOH were time-dependently observed. The amounts of these spin adducts were larger than those of PMNs stimulated by PMA. These results clearly show that MyA is more potent agent to prime human PMNs than PMA, in a point of view of not only O2·? but also ·OH production. This fact calls attention that too much intake of MyA that is known to be contained vegetable oils can lead to crippling effect through uncontrolled production of reactive oxygen species. PMID:19902021

  6. Toll-Like Receptor 1/2 Stimulation Induces Elevated Interleukin-8 Secretion in Polymorphonuclear Leukocytes Isolated from Preterm and Term Newborn Infants

    PubMed Central

    Thornton, Nathan L.; Cody, Mark J.; Yost, Christian C.

    2012-01-01

    Background Neonatal neutrophil dysfunction contributes to inflammatory tissue damage in newborn infants. Toll-like receptors (TLRs) activate the innate immune response through recognition of pathogen-associated molecular patterns. Expression and function of TLRs by neonatal neutrophils has not well been characterized. Objective: We hypothesized that, compared to polymorphonuclear leukocytes (PMNs) isolated from adults, neonatal PMNs isolated from either term or preterm infants express and release different levels of inflammatory cytokines and chemokines in response to stimulation with TLR1–9 agonists. Methods We stimulated PMNs isolated from preterm (n = 12) and term (n = 10) infants as well as adults (n = 10) with agonists recognized by TLRs1–9 and quantified chemokine and cytokine expression and secretion by ELISA and Luminex® multiplex quantification assay. Results Neonatal and adult PMNs stimulated with agonists recognized by TLRs1–9 differentially secrete inflammatory products. Signaling via TLR2 heterodimers is a potent mechanism for release of interleukin-8, a critical proinflammatory chemokine, by neonatal PMNs - a previously unrecognized facet of neonatal inflammation. Following TLR1/2 (PAM3CSK4) stimulation, interleukin-8 secretion by neonatal PMNs, whether term or preterm, substantially exceeds that of adult PMNs assayed in parallel. Conclusions These studies provide new insights relevant to the inflammatory biology of neonates, both term and preterm, and implicate exaggerated PMN recruitment in neonatal syndromes of dysregulated inflammation such as necrotizing enterocolitis or neonatal chronic lung disease. PMID:21952587

  7. Myeloid Derived Suppressor Cells (MDSCs) Are Increased and Exert Immunosuppressive Activity Together with Polymorphonuclear Leukocytes (PMNs) in Chronic Myeloid Leukemia Patients

    PubMed Central

    Giallongo, Cesarina; Parrinello, Nunziatina; Tibullo, Daniele; La Cava, Piera; Romano, Alessandra; Chiarenza, Annalisa; Barbagallo, Ignazio; Palumbo, Giuseppe A.; Stagno, Fabio; Vigneri, Paolo; Di Raimondo, Francesco

    2014-01-01

    Tumor immune tolerance can derive from the recruitment of suppressor cell population, including myeloid derived suppressor cells (MDSCs), able to inhibit T cells activity. We identified a significantly expanded MDSCs population in chronic myeloid leukemia (CML) patients at diagnosis that decreased to normal levels after imatinib therapy. In addition, expression of arginase 1 (Arg1) that depletes microenvironment of arginine, an essential aminoacid for T cell function, resulted in an increase in patients at diagnosis. Purified CML CD11b+CD33+CD14-HLADR- cells markedly suppressed normal donor T cell proliferation in vitro. Comparing CML Gr-MDSCs to autologous polymorphonuclear leukocytes (PMNs) we observed a higher Arg1 expression and activity in PMNs, together with an inhibitory effect on T cells in vitro. Our data indicate that CML cells create an immuno-tolerant environment associated to MDSCs expansion with immunosuppressive capacity mediated by Arg1. In addition, we demonstrated for the first time also an immunosuppressive activity of CML PMNs, suggesting a strong potential immune escape mechanism created by CML cells, which control the anti-tumor reactive T cells. MDSCs should be monitored in imatinib discontinuation trials to understand their importance in relapsing patients. PMID:25014230

  8. Functional and metabolic studies of polymorphonuclear leukocytes in the canine homologue of congenital Pelger-Huet Anomaly 

    E-print Network

    Browder, Elizabeth Jarratt

    1985-01-01

    Leukocytes in the Canine Homologue of Congenital Pelger-Huet Anomaly . (May 1985) Elizabeth Jarratt Browder, B. A. , Bay lor University; B. S. , Texas ASM University; D. V. M. , Texas A8M University Chairman of Advisory Committee: Dr. lan Tizard... are in order to the LARR staff for their help and moral support, to BLRC for art work and a great attitude even when deadlines were tight, and to Dr. Elizabeth Wilson for providing the kodachromes from which the photographs were taken. INTRODUCTION. TABLE...

  9. Modified Leukocyte Filter Removes Tumor Cells from the Salvaged Blood

    PubMed Central

    Zhang, Zhaohui; Zhang, Fengjiang; Gong, Lina; Sun, Kai; Zhang, Jie; Tang, Yumin; Jiang, Chunling; Liu, Jin

    2015-01-01

    Background Intraoperative blood salvage, an effective blood conservation strategy, has not been applied in onco-surgery, because of potential malignant cell contamination. In this study we tested effectiveness of a modified leukocyte depletion filter (M-LDF) for removal of tumor cells. Materials and Methods The effects of M-LDF and regular LDF on removal of cells (HepG2 cell line) were compared. The safety of M-LDF was tested with blood (collected and washed during onco-surgery), the salvaged blood mixed with tumor cells from the solid tumor of the same patient, or mixed with HepG2 cells (n=30 in each protocol). Cancer cells were identified by flow cytometry, culture and bioassay with and without filtration. Results M-LDF removed 5-log of HepG2 and nucleated cells, which was much higher than regular LDF, and cells were destroyed when they passed through M-LDF. Cytokeratin-positive cells in all samples were removed by M-LDF. Invasive growth adherent cells were found in most of unfiltered samples and 67% of the inoculated nude mice developed tumors in LDF-treated sample. Neither adherent cells nor nude mice developed tumors were found in M-LDF-treated samples. Discussion and Conclusion Since M-LDF can effectively remove and destroy cancer cells in the salvaged blood, it has great potential for clinical application. PMID:26098626

  10. Randomized trial comparing packed red cell blood transfusion with and without leukocyte depletion for gastrointestinal surgery

    Microsoft Academic Search

    Paul Ian Tartter; Kala Mohandas; Penny Azar; Jill Endres; Jess Kaplan; Morton Spivack

    1998-01-01

    BACKGROUND: Allogeneic transfusion is associated with postoperative infections that significantly prolong hospital stays and increase costs. Recent studies suggest that filtering leukocytes from blood prior to transfusion reduces the risk of postoperative infection associated with blood transfusion. We compared the incidence of postoperative infections, hospital stays, and hospital charges of gastrointestinal surgery patients transfused with packed red cells or leukocyte-depleted

  11. The manganese-salen compound EUK-134 and N-acetyl cysteine rescue from zinc- and paraquat-induced toxicity in rat polymorphonuclear leukocytes.

    PubMed

    Kumar, Ashutosh; Shukla, Smriti; Chauhan, Amit Kumar; Singh, Deepali; Pandey, Haushila Prasad; Singh, Chetna

    2015-04-25

    Oxidative stress is implicated in toxicant-induced inflammation leading to chronic diseases. Polymorphonuclear leukocytes (PMNs) offer the first line of defense against infection in the mammals and protect against inflammation-mediated pathological anomalies. Conversely, activated PMNs contribute to the oxidative stress-mediated damage and inflammation. The study aimed to investigate the status of oxidative stress and antioxidant defense system in the PMNs of rats treated with/without zinc (Zn) and/or paraquat (PQ) in the presence or absence of a synthetic superoxide dismutase/catalase mimetic, a manganese-salen compound-EUK-134 and/or a glutathione precursor, N-acetyl cysteine (NAC). While Zn and/or PQ elevated the total free radical generation, lipid peroxidation (LPO) and catalytic activity of myeloperoxidase (MPO), superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione S-transferase alpha 4-4 (GSTA4-4), a pronounced decrease in reduced glutathione (GSH) and glutathione reductase (GR) activity was also observed. Zn and/or PQ augmented the expression of metallothionein-I and II and GSTA4-4. Pre-treatment of EUK-134 or NAC alone altered the level of total free radical generation, LPO, GSH content and catalytic activity of MPO, SOD, GR and GPx and the expression of metallothionein I and II towards normalcy. The alterations were more pronounced in the PMNs of rats treated with EUK-134 and NAC in combination. Catalytic activity/expression of GSTA4-4 remained unchanged in the PMNs of EUK-134 or NAC treated rats. The results demonstrate that EUK-134 and NAC protect PMNs from the toxic effects of Zn and PQ in rats and also suggest that metallothioneins I/II might contribute to antioxidant defense under GSH depleted conditions. PMID:25724285

  12. Abnormalities of polymorphonuclear leukocyte function associated with a heritable deficiency of high molecular weight surface glycoproteins (GP138): common relationship to diminished cell adherence.

    PubMed Central

    Anderson, D C; Schmalstieg, F C; Arnaout, M A; Kohl, S; Tosi, M F; Dana, N; Buffone, G J; Hughes, B J; Brinkley, B R; Dickey, W D

    1984-01-01

    Investigations of polymorphonuclear leukocyte (PMN) function were performed in a 5-yr-old white female with delayed umbilical cord separation, impaired pus formation, and a severe defect of PMN chemotaxis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated an almost total deficiency of a high molecular weight glycoprotein(s) (GP138) in the granule and membrane fractions of the patient's cells, and NaB3H4-galactose oxidase labeling demonstrated the absence of a major glycoprotein complex on the surface of her PMNs. Monoclonal antibodies (MAb) were employed in flow cytometry experiments to demonstrate that two previously characterized glycoproteins (Mo1 and LFA1) were undetectable on the surface of the patient's PMNs and monocytes. Immunoprecipitation of 125I-labeled patient cells with subunit specific MAbs confirmed that the alpha-subunits of Mo1 (155 kD) and LFA1 (177 kD) and their common beta-subunit (94 kD) were totally deficient. Functional analyses of patient PMNs demonstrated severe impairment of adherence- and adhesion-dependent cell functions including spreading, aggregation, orientation in chemotactic gradients, antibody-dependent cellular cytotoxicity, and phagocytosis of particles (Oil-Red-0-paraffin, zymosan) selectively opsonized with C3-derived ligands. Patient PMNs demonstrated a normal capacity to rosette with IgG or C3b-coated sheep erythrocytes, but rosette formation with C3bi-coated erythrocytes was profoundly diminished. Adhesion-independent functions including shape change, N-formyl-methionyl-leucyl-3H-phenylalanine binding, and O-2 generation or secretion elicited by soluble stimuli were normal. Membrane fluidity, surface charge, and microtubule assembly were also normal. These findings provide new evidence that critical PMN surface glycoproteins are required to facilitate multiple adhesion-dependent cellular functions of the inflammatory response. Images PMID:6746906

  13. Structural analysis of the cellular constituents of a fresh clinical isolate of Staphylococcus aureus, and their role in the interaction between the organisms and polymorphonuclear leukocytes in the presence of serum factors.

    PubMed Central

    Karakawa, W W; Young, D A; Kane, J A

    1978-01-01

    The in vitro interaction of a fresh clinical isolate of Staphylococcus aureus and polymorphonuclear leukocytes was investigated. The importance of the various cellular constituents as host immunological factors was analyzed, and the results suggested that two components, namely, an acidic polysaccharide consisting of a predominance of aminogalacturonic acid and a strain-specific mucopeptide complex, may be involved in impeding in vitro opsonization of the organism by leukocytes. Immunochemical analysis indicated that the acidic polysaccharide possessed the same immunodominant aminogalacturonic acid residues as the antiphagocytic acidic antigen of the encapsulated prototype Scott strain. Antisera derived from rabbits immunized with strain D contained two types of opsonins, those with acidic polymer specificity and those with mucopeptide complex specificity. Images PMID:689733

  14. Validation of automated blood cell counter for the determination of polymorphonuclear cell count in the ascitic fluid of cirrhotic patients with or without spontaneous bacterial peritonitis

    Microsoft Academic Search

    Stefania Angeloni; Giorgia Nicolini; Manuela Merli; Francesca Nicolao; Giorgio Pinto; Teresa Aronne; Adolfo Francesco Attili; Oliviero Riggio

    2003-01-01

    ObjectivePolymorphonuclear (PMN) cell count in ascitic fluid is the most useful test for the diagnosis of spontaneous bacterial peritonitis (SBP). We evaluated the validity of an automated blood cell counter for the PMN determination in ascitic fluid by comparing it with the traditional hematologic method with a light microscope in a manual counting chamber.

  15. Severe microvascular injury induced by lysosomal releasates of human polymorphonuclear leukocytes. Increase in vasopermeability, hemorrhage, and microthrombosis due to degradation of subendothelial and perivascular matrices.

    PubMed Central

    Movat, H. Z.; Wasi, S.

    1985-01-01

    The purpose of this study was to assess the nature of the lesions in the microcirculation of the dermis of rabbits induced with lysosomal releasates of human polymorphonuclear leukocytes (PMNs). No attempt was made in the studies presented in this publication to deal with the offending agent in the releasate. Four parameters of microvascular injury were quantitated: increase in vascular permeability with 125I-labeled serum albumin, hemorrhage with 59Fe-labeled erythrocytes, accumulation (aggregation) of platelets with 111In-labeled platelets. In one experiment accumulation of 51Cr-PMNs was investigated. The lysosomal releasate induced a rapid increase in vasopermeability, but both hemorrhage and exudate formation peaked 1 hour after intradermal injection. Platelet accumulation was also demonstrable in these lesions, and microthrombosis was a very prominent feature. The microvascular injury, including microthrombosis, could be elicited also in animals rendered leukopenic with nitrogen mustard. Simultaneous injection of prostaglandin E2 with the releasate enhanced the microvascular injury. The morphologic changes in the microcirculation of the rabbit's dermis were assessed in lesions 5 minutes to 5 hours old. Several changes were encountered, primarily in the wall of venules and small veins and to a lesser degree in small arteries and capillaries. Ultrastructurally very early lesions (up to 15 minutes) had gaps or spaces in the endothelium, resembling those induced by mediators such as histamine or bradykinin. Older lesions were different, quite characteristic, and represent the hallmark of these lesions. Lysis and disappearance of vascular basement membrane, of perivascular collagen, and of the internal elastic lamina were a frequent finding, best demonstrable when microthrombi did not abut on vessel walls. Cellular components of vessels (endothelium, pericytes, smooth muscle) showed fragmentation, leading to complete disappearance of cellular elements. These lesions were usually walled off by platelet aggregates and fibrin. At times microthrombi occluded an entire vessel. These changes were interpreted as hemostasis. The mild accumulation of PMNs at the site of injury did not contribute significantly to the microvascular injury. The findings indicate that the unique changes in the microcirculation, not described before, may occur quite frequently, when the microvascular injury is elicited primarily by release of lysosomal constituents by phagocytic or nonphagocytic stimuli. One can conclude that the hallmark of this type of injury is disappearance of basement membrane followed secondarily by disintegration of the vascular wall, followed in turn by hemo Images Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12 Figure 13 PMID:3907363

  16. CYP2E1-mediated oxidative stress regulates HO-1 and GST expression in maneb- and paraquat-treated rat polymorphonuclear leukocytes.

    PubMed

    Ahmad, Israr; Shukla, Smriti; Singh, Deepali; Chauhan, Amit Kumar; Kumar, Vinod; Singh, Brajesh Kumar; Patel, Devendra Kumar; Pandey, Haushila Prasad; Singh, Chetna

    2014-08-01

    Cytochrome P4502E1 (CYP2E1), glutathione-S-transferase A4-4 (GSTA4-4), and inducible nitric oxide synthase (iNOS) are implicated in maneb- and paraquat-induced toxicity leading to various pathological conditions. The study aimed to investigate the role of CYP2E1 in maneb- and paraquat-induced oxidative stress in rat polymorphonuclear leukocytes (PMNs) and its crosstalk with iNOS-mediated nitrosative stress and GSTA4-4-linked protective effect, if any and their consequent links with the nuclear factor erythoid 2-related factor 2 (Nrf2) activation and heme oxygenase-1 (HO-1) expression. Rats were treated with/without maneb and/or paraquat for 1, 2, and 3 weeks along with vehicle controls. Subsets of rats were also treated with diallyl sulfide (DAS) or aminoguanidine (AG) along with the respective controls. Maneb and paraquat augmented the reactive oxygen species (ROS), lipid peroxidation (LPO) and 4-hydroxy nonenal (4-HNE) contents, and superoxide dismutase (SOD) activity in the PMNs. However, maneb and paraquat attenuated the reduced glutathione (GSH) level and the expression/activity of total GST and GST-pi. Maneb and paraquat increased the expression/activity of CYP2E1, GSTA4-4, iNOS, Nrf2 and HO-1, and nitrite content. CYP2E1 inhibitor, DAS noticeably alleviated maneb- and paraquat-induced ROS, LPO, 4-HNE, SOD, Nrf2 and HO-1, GST, GSH, and GST-pi while iNOS, nitrite content and GSTA4-4 levels were unchanged. Conversely, AG, an iNOS inhibitor, attenuated maneb- and paraquat-directed changes in nitrite, LPO, iNOS but it did not alter ROS, GSH, SOD, GST, GST-pi, Nrf2, HO-1, CYP2E1, and GSTA4-4. The results demonstrate that CYP2E1 induces iNOS-independent free radical generation and subsequently modulates the Nrf2-dependent HO-1 and 4-HNE-mediated GST expression in maneb- and paraquat-treated PMNs. PMID:24771067

  17. Microsatellite Instability in the Peripheral Blood Leukocytes of HNPCC Patients

    PubMed Central

    Coolbaugh-Murphy, Mary I.; Xu, Jing-Ping; Ramagli, Louis S.; Ramagli, Brian C.; Brown, Barry W.; Lynch, Patrick M.; Hamilton, Stanley R.; Frazier, Marsha L.; Siciliano, Michael J.

    2012-01-01

    Most hereditary nonpolyposis colorectal cancer (HNPCC) patients inherit a defective allele of a mismatch repair (MMR) gene, usually MLH1 or MSH2, resulting in high levels of microsatellite instability (MSIH) in the tumors. Presence of MSI in the normal tissues of mutation carriers has been controversial. Here we directly compare MSI in the peripheral blood leukocyte (PBL) DNA of seven HNPCC patients carrying different types of pathogenic MMR mutations in MLH1 and MSH2 genes with the PBL DNA of normal age-matched controls and of patients with sporadic colorectal cancer (SCRC). Small pool PCR (SP-PCR) was used studying three microsatellite loci for at least 100 alleles each in most samples. The average frequencies of mutant microsatellite fragments in each HNPCC patient (0.04–0.24) were significantly higher (p<0.01) relative to their age-matched normal controls with mutant frequencies (MF) from 0.00 to 0.06, or SCRC patients (MF from 0.01–0.03). The data support the conclusions that higher MF in the PBL DNA of HNPCC patients is real and reproducible, may vary in extent according to the type of germline MMR mutation and the age of the individual, and provide a possible genetic explanation for anticipation in HNPCC families. PMID:20052760

  18. Obesity related methylation changes in DNA of peripheral blood leukocytes

    PubMed Central

    2010-01-01

    Background Despite evidence linking obesity to impaired immune function, little is known about the specific mechanisms. Because of emerging evidence that immune responses are epigenetically regulated, we hypothesized that DNA methylation changes are involved in obesity induced immune dysfunction and aimed to identify these changes. Method We conducted a genome wide methylation analysis on seven obese cases and seven lean controls aged 14 to 18 years from extreme ends of the obesity distribution and performed further validation of six CpG sites from six genes in 46 obese cases and 46 lean controls aged 14 to 30 years. Results In comparison with the lean controls, we observed one CpG site in the UBASH3A gene showing higher methylation levels and one CpG site in the TRIM3 gene showing lower methylation levels in the obese cases in both the genome wide step (P = 5 × 10-6 and P = 2 × 10-5 for the UBASH3A and the TRIM3 gene respectively) and the validation step (P = 0.008 and P = 0.001 for the UBASH3A and the TRIM3 gene respectively). Conclusions Our results provide evidence that obesity is associated with methylation changes in blood leukocyte DNA. Further studies are warranted to determine the causal direction of this relationship as well as whether such methylation changes can lead to immune dysfunction. See commentary: http://www.biomedcentral.com/1741-7015/8/88/abstract PMID:21176133

  19. TRANSLATIONAL PHYSIOLOGY Whole blood and leukocyte RNA isolation for gene expression analyses

    E-print Network

    Richardson, David

    TRANSLATIONAL PHYSIOLOGY Whole blood and leukocyte RNA isolation for gene expression analyses of Florida College of Medicine, Gainesville, Florida; 3 Stanford Genome Technology Center, Palo Alto of Medicine, St. Louis, Missouri; 6 Department of Chemical Engineering, Massachusetts Institute of Technology

  20. Reduction of HTLV-I-infected cells in blood by leukocyte filtration.

    PubMed

    Al, E J; Visser, S C; Broersen, S M; Stienstra, S; Huisman, J G

    1993-12-01

    Leukocyte filtration was performed with HTLV-I-infected blood and with blood supplemented with cultured HTLV-I-transformed cells. Reduction of infectivity upon leukocyte filtration was determined by the polymerase chain reaction (PCR) using primers indicative for the HTLV-I-pol and tax genes. Two different commercially available filters were used: a column-shaped cellulose acetate and a flat-bed polyester filter. Both filters yielded reduction of at least 3 10logs for cultured HTLV-I-infected cells. When blood from HTLV-I-infected individuals was used for filtration, the number of infected cells was reduced by 1-3 10logs. Although filtered blood as yet cannot be regarded as safe, it is concluded that leukocyte filtration of HTLV-I-infected blood potentially contributes to reducing the spread of HTLV-I by blood transfusion. PMID:8280813

  1. Blood immunometabolic indices and polymorphonuclear neutrophil function in peripartum dairy cows are altered by level of dietary energy prepartum.

    PubMed

    Graugnard, D E; Bionaz, M; Trevisi, E; Moyes, K M; Salak-Johnson, J L; Wallace, R L; Drackley, J K; Bertoni, G; Loor, J J

    2012-04-01

    Cows experience some degree of negative energy balance and immunosuppression around parturition, making them vulnerable to metabolic and infectious diseases. The effect of prepartum feeding of diets to meet (control, 1.34 Mcal/kg of dry matter) or exceed (overfed, 1.62 Mcal/kg of dry matter) dietary energy requirements was evaluated during the entire dry period (?45 d) on blood polymorphonuclear neutrophil function, blood metabolic and inflammatory indices, and milk production in Holstein cows. By design, dry matter intake in the overfed group (n=9) exceeded energy requirements during the prepartum period (-4 to -1 wk relative to parturition), resulting in greater energy balance when compared with the control group (n=10). Overfed cows were in more negative energy balance during wk 1 after calving than controls. No differences were observed in dry matter intake, milk yield, and milk composition between diets. Although nonesterified fatty acid concentration pre- (0.138 mEq/L) and postpartum (0.421 mEq/L) was not different between diets, blood insulin concentration was greater in overfed cows prepartum (16.7 ?IU/mL) compared with controls pre- and postpartum (?3.25 ?IU/mL). Among metabolic indicators, concentrations of urea (4.63 vs. 6.38 mmol/L), creatinine (100 vs. 118 ?mol/L), and triacylglycerol (4.0 vs. 8.57 mg/dL) in overfed cows were lower prepartum than controls. Glucose was greater pre- (4.24 vs. 4.00 mmol/L) and postpartum (3.49 vs. 3.30 mmol/L) compared with control cows. Among liver function indicators, the concentration of bilirubin increased by 2 to 6 fold postpartum in control and overfed cows. Phagocytosis capacity of polymorphonuclear neutrophils was lower prepartum in overfed cows (32.7% vs. 46.5%); phagocytosis in the control group remained constant postpartum (50%) but it increased at d 7 in the overfed group to levels similar to controls (48.4%). Regardless of prepartum diet, parturition was characterized by an increase in nonesterified fatty acid and liver triacylglycerol, as well as blood indices of inflammation (ceruloplasmin and haptoglobin), oxidative stress (reactive oxygen metabolites), and liver injury (glutamic oxaloacetic transaminase). Concentrations of the antioxidant and anti-inflammatory compounds vitamin A, vitamin E, and ?-carotene decreased after calving. For vitamin A, the decrease was observed in overfed cows (47.3 vs. 27.5 ?g/100 mL). Overall, overfeeding energy and higher energy status prepartum led to the surge of insulin and had a transient effect on metabolism postpartum. PMID:22459823

  2. Flow cytometry follow-up analysis of peripheral blood leukocyte subpopulations in calves experimentally infected with field isolates of Mycoplasma bovis.

    PubMed

    Dudek, Katarzyna; Bednarek, Dariusz; Szacawa, Ewelina; Rosales, Ruben S; Ayling, Roger D

    2015-06-01

    Changes in peripheral blood leukocyte subpopulations were investigated in calves challenged intratracheally with three different Mycoplasma bovis isolates in Groups E1, E2, and E3. The controls received a placebo. Blood samples were collected before challenge and then at days 1 to 7, 14, 21 and 28. White blood cells (WBC), polymorphonuclear leukocytes (PMNLs), lymphocytes (LYMs), monocytes, eosinophils and basophils (mid-size cells, MID), as well as CD2(+), CD4(+), CD8(+), WC4(+) lymphocyte subsets with CD4:CD8 ratio were also analysed. A transient increase of WBC and PMNLs in all challenged calves was observed on day 1. Increased LYM counts were observed in E1 throughout the study, whereas in E2 the LYM counts were higher only between days 14 and 28, and consistently lower in E3. The MID count had broadly comparable values for all groups. Stimulation of the CD2(+) response was observed in E2 and E3 in contrast with E1 which had a lower CD2(+) throughout. The CD4(+) response was dominant in E1 and E2, whereas in E3 a parallel CD4(+) and CD8(+) stimulation was observed. The B-cell response (WC4(+)) and an increased CD4:CD8 ratio was most apparent in E1. The main host responses to M. bovis infections are a stimulation of CD4(+) cells and an enhancement of the WC4(+) response. PMID:26051255

  3. Validation of automated blood cell counter for the determination of polymorphonuclear cell count in the ascitic fluid of cirrhotic patients with or without spontaneous bacterial peritonitis

    Microsoft Academic Search

    Stefania Angeloni; Giorgia Nicolini; Manuela Merli; Francesca Nicolao; Giorgio Pinto; Teresa Aronne; Adolfo Francesco Attili; Oliviero Riggio

    2003-01-01

    OBJECTIVE:Polymorphonuclear (PMN) cell count in ascitic fluid is the most useful test for the diagnosis of spontaneous bacterial peritonitis (SBP). We evaluated the validity of an automated blood cell counter for the PMN determination in ascitic fluid by comparing it with the traditional hematologic method with a light microscope in a manual counting chamber.METHODS:A total of 130 ascitic fluid samples

  4. Antitumor effect induced by a hot water extract of Chlorella vulgaris (CE): Resistance to meth-A tumor growth mediated by CE-induced polymorphonuclear leukocytes

    Microsoft Academic Search

    Fumiko Konishi; Kuniaki Tanaka; Kunisuke Himeno; Kazuto Taniguchi; Kikuo Nomoto

    1985-01-01

    When a hot water extract of Chlorella vulgaris (CE) was injected into the peritoneal cavity of BALB\\/c mice inoculated with syngeneic Meth-A tumor cells, the survival times were strikingly prolonged. Furthermore, peritoneal exudate cells (PEC) rich in polymorphonuclear cells (PMN) obtained from normal mice 24 h after CE injection exhibited an antitumor effect in a Winn-type assay using normal recipients.

  5. Differential leukocyte counting and immunophenotyping in cryopreserved ex vivo whole blood.

    PubMed

    Nemes, Elisa; Kagina, Benjamin M N; Smit, Erica; Africa, Hadn; Steyn, Marcia; Hanekom, Willem A; Scriba, Thomas J

    2015-02-01

    Absolute cell counts are typically measured in fresh samples, but this is impractical in large field studies. We compared quantification of leukocyte proportions and absolute counts using reference real-time methods (stain and lyse/no-wash (LNW) or hematology analyser) with a novel assay that allows long-term cryopreservation of fixed leukocytes for later counting (DLC-ICE: differential leukocyte count and immunophenotype in cryopreserved ex vivo whole blood). For the LNW method, whole blood (WB) was stained with fluorescent antibodies, then erythrocytes were lysed, and leukocytes fixed prior to flow cytometry. Alternatively, our novel DLC-ICE method entailed erythrocyte lysis and leukocyte fixation, cryopreservation and later staining of permeabilized cells prior to flow cytometry. Outcomes were proportions and absolute counts of granulocytes, lymphocytes, monocytes, T cells, B cells, and activated T cells within the leukocyte population. We also compared leukocyte subset counts in fresh WB from 51 healthy infants measured by hematology analyser at a rural clinical site or by DLC-ICE method after 2 years of cryopreservation. We observed excellent agreement and strong correlations between absolute counts or cell proportions measured by the LNW and DLC-ICE methods on fresh WB from 10 healthy adults. Compared to LNW, DLC-ICE yielded similar or brighter staining even after cryopreservation. Duration of cryopreservation, assessed monthly for 1 year, had little effect on cell enumeration: median coefficients of variation were below 15% for all outcomes. Under field site conditions, we observed strong correlations between infant leukocyte numbers measured in fresh samples by hematology analyser and those measured by DLC-ICE up to 2 years of cryopreservation. Our novel DLC-ICE method allows accurate flow cytometric quantification of cell subsets from fixed WB even after long-term cryopreservation. This method is ideal for batched, retrospective analysis of samples from large field studies, or when advanced flow cytometry equipment is not available for clinical research purposes. © 2014 International Society for Advancement of Cytometry. PMID:25515205

  6. [Selectivity of accumulation of chlorine e6 derivatives in blood leukocytes].

    PubMed

    Savitski?, V P; Zorin, V P

    2003-01-01

    The uptake of chlorine e6 derivatives by peripheral blood leukocytes was studied using a fluorescence-activated sorter. The analysis showed that the order of pigment uptake by leukocyte populations is the following: granulocytes > or = monocytes > lymphocytes. It was found that the accumulation of the pigments in the cell significantly varied. The level of chlorine e6 dimethyl ester accumulated by cells was found to be 15 times higher than that of chlorine e6. It was assumed that the differences in pigment uptake by different types of blood cells are due to structural and morphological features of leukocytes. The data obtained may be useful in developing the new methods of photodynamic therapy. PMID:12630115

  7. LYSIS OF ANTIBODY COATED CHICKEN ERYTHROCYTES BY A NON-LYMPHOCYTE PIG BLOOD LEUKOCYTE

    E-print Network

    Paris-Sud XI, Université de

    LYSIS OF ANTIBODY COATED CHICKEN ERYTHROCYTES BY A NON-LYMPHOCYTE PIG BLOOD LEUKOCYTE B. CHARLEY H : erythrocyte-antibody. Koren et al., 1978 ; Wardley et al., 1976). Dif- ferent cell types, all of which bearing communication, using a con- ventionnal slchromium release test with label- led chicken erythrocytes sensitized

  8. Phagocytic and bactericidal activities of leukocytes in whole blood from atomic bomb survivors

    SciTech Connect

    Sasagawa, S.; Yoshimoto, Y.; Toyota, E.; Neriishi, S.; Yamakido, M.; Matsuo, M.; Hosoda, Y.; Finch, S.C. (Hiroshima Univ. (Japan))

    1990-10-01

    This study evaluated the phagocytic and bactericidal activities of peripheral blood leukocytes from Hiroshima and Nagasaki atomic bomb survivors for Staphylococcus aureus. The data were analyzed by multiple linear regression for age, sex, radiation exposure, city of exposure, and neutrophil counts. No significant radiation effect was observed for either blood phagocytic or bactericidal activities. The only significant variable for these functions was the neutrophil count.

  9. Leukocyte accumulation in graft blood vessels during self-limiting acute rejection of rat kidneys.

    PubMed

    Zakrzewicz, Anna; Wilhelm, Jochen; Blöcher, Sonja; Wilczynska, Joanna; Wilker, Sigrid; Dietrich, Hartmut; Weimer, Rolf; Padberg, Winfried; Grau, Veronika

    2011-05-01

    During self-limiting acute rejection preceding chronic vasculopathy, large amounts of leukocytes, predominantly monocytes, interact with the endothelium of renal allografts. We aim to characterize them and to identify targets for functional and interventional studies. Leukocytes were harvested by vascular perfusion from Fischer 344 to Lewis renal allografts or Lewis isografts, followed by flow cytometry, quantitative RT-PCR and genome-wide transcriptional profiling. Leukocyte accumulation peaked in allografts on day 9. The percentage of monocytes expressing MHC class II and CD161 was increased whereas CD4, CD11a, CD43, and CD71 expression remained unchanged. IFN-?, IL-1?, IL-2, IL-10, TNF-?, and iNOS mRNA increased in allograft leukocytes but IL-4, IL-6, IL-12, TGF-?, and tissue factor did not. During acute rejection, 1783 genes were differentially expressed. In conclusion, graft blood leukocytes display a unique state of partial activation during self-limiting rejection. Numerous differentially expressed genes deserve further investigation as potential factors in deciding the fate of the allograft. PMID:21035231

  10. Laser-induced priming of human blood leukocytes

    NASA Astrophysics Data System (ADS)

    Chichuk, Tatyana V.; Stranadko, Eugeny P.; Strashkevich, I. A.; Klebanov, Gennady I.

    1999-12-01

    We investigated the influence of He-Ne ((lambda) equals 632.8 nm) laser irradiation (LI) on a functional activity of human blood leucocytes. The method of luminol-dependent chemiluminescence with the zymosan-activated phagocytes was used. The leucocytes were irradiated without and in the presence of autologic human blood plasma, containing of the endogenous (porphyrins) and/or exogenous (phthalocyanine) photosensitizers. The LI initiated a priming of the leucocytes. Priming revealed itself after the activation of the phagocytes by zymosan. The changes of the calcium concentration in leucocytes cytoplasm were studied too. Fluorimetric method with Fura-2AM was used for this. The laser irradiation initiated the changes of the calcium concentration in the leucocytes cytoplasm. All the investigating parameters depended on the irradiation dose and on the concentration of photosensitizers. The results of this work allowed to formulate the main theses of the free radical mechanism of the low intensive laser irradiation action on human blood leucocytes.

  11. Reduction of the surface charge of blood polymorphonuclear cells by rheumatoid sera and heat induced aggregated human IgG (HAGG).

    PubMed Central

    Brown, K A; McCarthy, D; Perry, J D; Dumonde, D C

    1988-01-01

    Cell electrophoresis identifies two main subpopulations of blood polymorphonuclear cells (PMNs), which in terms of the speed of their anodic migration are referred to as the fast and slow population. When blood PMNs from normal healthy subjects were incubated in medium containing 20% RA serum there was a decrease in the percentage of fast cells with a corresponding increase in the slow population that was directly related to the levels of circulating immune complexes present in the sera. Similar results were obtained when using heat induced aggregated human IgG (HAGG) or Candida albicans instead of RA serum. The 'slowing' effect of HAGG, which was transient and time dependent, appeared to be due to its internalisation by the PMNs. These results suggest that in RA the large number of blood PMNs with a low surface charge (i.e., the slow population) may arise as a result of the constant interaction of these cells with circulating immune complexes. PMID:3291790

  12. Effect of replacing a high linoleate oil with a low linoleate, high alpha-linolenate oil, as compared with supplementing EPA or DHA, on reducing lipid mediator production in rat polymorphonuclear leukocytes.

    PubMed

    Ohhashi, K; Takahashi, T; Watanabe, S; Kobayashi, T; Okuyama, H; Hata, N; Misawa, Y

    1998-06-01

    The fatty acid composition of rat polymorphonuclear leukocytes (PMN) was modified by diets supplemented with a high linoleate (LA) safflower oil (76% LA), mixtures of eicosapentaenoate (EPA) and safflower oil (EPA(20) containing 20% EPA and 61% LA, EPA(40) containing 40% EPA and 46% LA), mixtures of docosahexaenoate (DHA) and safflower oil (DHA(20) containing 20% DHA and 61% LA, DHA(40) containing 40% DHA and 46% LA) or a high alpha-linolenate (alpha-LNA) perilla oil (57% alpha-LNA and 13% LA), and then lipid mediator production in casein-induced peritoneal PMN were compared. EPA and DHA were relatively ineffective in reducing platelet-activating factor (PAF) production; a statistically significant reduction was observed only in the DHA(40) group. In contrast, perilla oil reduced PAF production by 50% as compared with safflower oil. Arachidonate (AA) in the PAF precursor, 1-alkyl-2-acyl-glycerophosphocholine, was roughly correlated with PAF production, but EPA and DHA in the precursor lipid were relatively unrelated. On the other hand, both PGE2 and LTB4 production correlated positively with AA and negatively with EPA and DHA in PMN phospholipids; EPA tended to be somewhat more effective than DHA in reducing PGE2 and LTB4 formation; the activity of perilla oil was no less than EPA(20). Thus, replacing safflower oil with perilla oil was no less effective than supplementing safflower oil with EPA or DHA (at 40% of total fatty acids) in reducing lipid mediator production in rat PMN. PMID:9657037

  13. Possible mechanisms for the differential effects of high linoleate safflower oil and high alpha-linolenate perilla oil diets on platelet-activating factor production by rat polymorphonuclear leukocytes.

    PubMed

    Oh-hashi, K; Takahashi, T; Watanabe, S; Kobayashi, T; Okuyama, H

    1997-12-01

    As compared with high dietary linoleate safflower oil, high dietary alpha-linolenate perilla oil decreased platelet-activating factor (PAF) production by nearly half in calcium ionophore (CaI)-stimulated rat polymorphonuclear leukocytes (PMN). In the CaI-stimulated PMN from the perilla oil group, the accumulated amount of arachidonate (AA) plus eicosapentaenoate (EPA) was 30% less and that of lyso-PAF was 50% less, indicating that the decreased availability of lyso-PAF is a factor contributing to the relatively low PAF production. Consistently, eicosatetraynoic acid (ETYA), a dual inhibitor of cyclooxygenase and lipoxygenase, increased free fatty acids (FFA) and decreased PAF production possibly by decreasing the availability of lyso-PAF. Although, leukotrienes (LTs) have been proposed to stimulate PAF production synergistically, a potent LTB4 receptor antagonist, ONO-4057, decreased the formation of free fatty acids and LTB4, but stimulated PAF production somewhat, indicating that LTB4 may not stimulate PAF production in PMN. Lysophospholipid-induced transacylase (CoA-independent transacylase) activity in PMN homogenates was 25-30% lower in the perilla oil group but no significant differences were observed in the lyso-PAF acetyltransferase and PAF acetylhydrolase activities between the two dietary groups. Thus, decreased transacylase activity is another factor associated with the relatively low PAF production in the perilla oil group. PMID:9524928

  14. Effect of propofol on adhesion of activated platelets to leukocytes in human whole blood

    Microsoft Academic Search

    Lothar de Rossi; Martina Wessiepe; Wolfgang Buhre; Ralf Kuhlen; Gabriele Hutschenreuter; Rolf Rossaint

    2003-01-01

    Objective: To investigate the effect of propofol and its solvent Intralipid on the adhesion of activated platelets to leukocytes in vitro. Design and setting: Prospective study in an experimental laboratory. Participants: Sixteen healthy volunteers. Interventions: Whole blood was incubated for 60 min with propofol (4, 40 µg\\/ml), an equal volume of Intralipid 10% or phosphate-buffered saline (PBS). After stimulation with adenosine-5-diphosphate (ADP)

  15. Measurement of Phosphatidylserine Exposure in Leukocytes and Platelets by Whole-Blood Flow Cytometry with Annexin V

    Microsoft Academic Search

    Jonathan F. Tait; Christina Smith

    1999-01-01

    Phosphatidylserine (PS) exposure serves as a procoagulant stimulus and a signal for phagocytic clearance of apoptotic cells. In order to measure PS exposure in blood cells, we developed a flow-cytometric procedure to measure annexin V binding to leukocytes and platelets in whole-blood samples. Leukocytes were identified by CD45 and side-scatter gating, and platelets by CD61 and side- scatter gating. The

  16. Measurement of Phosphatidylserine Exposure in Leukocytes and Platelets by Whole-Blood Flow Cytometry with Annexin V

    Microsoft Academic Search

    Jonathan F Tait; Christina Smith; Brent L Wood

    1999-01-01

    ABSTRACT: Phosphatidylserine (PS) exposure serves as a procoagulant stimulus and a signal for phagocytic clearance of apoptotic cells. In order to measure PS exposure in blood cells, we developed a flow-cytometric procedure to measure annexin V binding to leukocytes and platelets in whole-blood samples. Leukocytes were identified by CD45 and side-scatter gating, and platelets by CD61 and side-scatter gating. The

  17. Quantitation of cytomegalovirus (CMV) DNA in leukocytes of human immunodeficiency virus-infected subjects with and without CMV disease by using PCR and the SHARP Signal Detection System.

    PubMed Central

    Boivin, G; Handfield, J; Murray, G; Toma, E; Lalonde, R; Lazar, J G; Bergeron, M G

    1997-01-01

    We report the development of a simple and rapid PCR assay for quantitation of the cytomegalovirus (CMV) DNA load in polymorphonuclear leukocytes. Using this system, a very good correlation was found between a high number of CMV copies in the blood and the presence of CMV disease in subjects with AIDS. PMID:9003635

  18. Shorter telomere length in peripheral blood leukocytes is associated with childhood autism

    PubMed Central

    Li, Zongchang; Tang, Jinsong; Li, Hong; Chen, Shan; He, Ying; Liao, Yanhui; Wei, Zhen; Wan, Guobin; Xiang, Xi; Xia, Kun; Chen, Xiaogang

    2014-01-01

    Telomeres are protective chromosomal structures that play a key role in preserving genomic stability. Epidemiologic studies have shown that the abnormal telomere length in leukocytes is associated with some mental disorders and age-related diseases. However, the association between leukocyte telomere length and autism has not been investigated. Here we investigated the possible association between relative telomere length (RTL) in peripheral blood leukocytes and childhood autism by using an established real-time polymerase chain reaction method. We observed significantly shorter RTL in patients with childhood autism than in controls (p = 0.006). Individuals with shorter RTL had a significantly increased presence of childhood autism compared with those who had long RTL. In patients, we found that family training interventions have a significant effect on telomere length (P = 0.012), but no correlations between RTL and clinical features (paternal age, maternal age, age of onset, illness of duration, CARS score and ABC score) were observed in this study. These results provided the first evidence that shorter leukocytes telomere length is significantly associated with childhood autism. The molecular mechanism underlying telomere length may be implicated in the development of autism. PMID:25399515

  19. Flow cytofluorometric assay of human whole blood leukocyte DNA degradation in response to Yersinia pestis and Staphylococcus aureus

    NASA Astrophysics Data System (ADS)

    Kravtsov, Alexander L.; Grebenyukova, Tatyana P.; Bobyleva, Elena V.; Golovko, Elena M.; Malyukova, Tatyana A.; Lyapin, Mikhail N.; Kostyukova, Tatyana A.; Yezhov, Igor N.; Kuznetsov, Oleg S.

    2001-05-01

    Human leukocytes containing less than 2C DNA per cell (damaged or dead cells) were detected and quantified by flow cytometry and DNA-specific staining with ethidium bromide and mithramycin in whole blood infected with Staphylococcus aureus or Yersinia pestis. Addition of live S. aureus to the blood (100 microbe cells per one leukocyte) resulted in rapid degradation of leukocyte DNA within 3 to 6 hours of incubation at 37 degree(s)C. However, only about 50 percent cells were damaged and the leukocytes with the intact genetic apparatus could be found in the blood for a period up to 24 hours. The leukocyte injury was preceded by an increase of DNA per cell content (as compared to the normal one) that was likely to be connected with the active phagocytosis of S. aureus by granulocytes (2C DNA of diploid phagocytes plus the all bacterial DNA absorbed). In response to the same dose of actively growing (at 37 degree(s)C) virulent Y. pestis cells, no increase in DNA content per cell could be observed in the human blood leukocytes. The process of the leukocyte DNA degradation started after a 6-hour incubation, and between 18 to 24 hours of incubation about 90 percent leukocytes (phagocytes and lymphocytes) lost their specific DNA fluorescence. These results demonstrated a high potential of flow cytometry in comparative analysis in vitro of the leukocyte DNA degradation process in human blood in response to bacteria with various pathogenic properties. They agree with the modern idea of an apoptotic mechanism of immunosuppression in plague.

  20. Biomarkers measured in buccal and blood leukocyte DNA as proxies for colon tissue global methylation.

    PubMed

    Ashbury, Janet E; Taylor, Sherryl A; Tse, M Yat; Pang, Stephen C; Louw, Jacob A; Vanner, Stephen J; King, Will D

    2014-01-01

    There is increasing interest in clarifying the role of global DNA methylation levels in colorectal cancer (CRC) etiology. Most commonly, in epidemiologic studies, methylation is measured in DNA derived from blood leukocytes as a proxy measure of methylation changes in colon tissue. However, little is known about the correlations between global methylation levels in DNA derived from colon tissue and more accessible tissues such as blood or buccal cells. This cross-sectional study utilized DNA samples from a screening colonoscopy population to determine to what extent LINE-1 methylation levels (as a proxy for genome-wide methylation) in non-target tissue (e.g., blood, buccal cells) reflected methylation patterns of colon mucosal tissue directly at risk of developing CRC. The strongest Pearson correlation was observed between LINE-1 methylation levels in buccal and blood leukocyte DNA (r = 0.50; N = 67), with weaker correlations for comparisons between blood and colon tissue (r = 0.36; N = 280), and buccal and colon tissue (r = 0.27; N = 72). These findings of weak/moderate correlations have important implications for interpreting and planning future investigations of epigenetic markers and CRC risk. PMID:24959316

  1. Effect of prenatal arsenic exposure on DNA methylation and leukocyte subpopulations in cord blood

    PubMed Central

    Kile, Molly L; Houseman, E Andres; Baccarelli, Andrea A; Quamruzzaman, Quazi; Rahman, Mahmuder; Mostofa, Golam; Cardenas, Andres; Wright, Robert O; Christiani, David C

    2014-01-01

    Prenatal arsenic exposure is associated with increased risk of disease in adulthood. This has led to considerable interest in arsenic’s ability to disrupt fetal programming. Many studies report that arsenic exposure alters DNA methylation in whole blood but these studies did not adjust for cell mixture. In this study, we examined the relationship between arsenic in maternal drinking water collected ? 16 weeks gestational age and DNA methylation in cord blood (n = 44) adjusting for leukocyte-tagged differentially methylated regions. DNA methylation was quantified using the Infinium HumanMethylation 450 BeadChip array. Recursively partitioned mixture modeling examined the relationship between arsenic and methylation at 473,844 CpG sites. Median arsenic concentration in water was 12 µg/L (range < 1- 510 µg/L). Log10 arsenic was associated with altered DNA methylation across the epigenome (P = 0.002); however, adjusting for leukocyte distributions attenuated this association (P = 0.013). We also observed that arsenic had a strong effect on the distribution of leukocytes in cord blood. In adjusted models, every log10 increase in maternal drinking water arsenic exposure was estimated to increase CD8+ T cells by 7.4% (P = 0.0004) and decrease in CD4+ T cells by 9.2% (P = 0.0002). These results show that prenatal exposure to arsenic had an exposure-dependent effect on specific T cell subpopulations in cord blood and altered DNA methylation in cord blood. Future research is needed to determine if these small changes in DNA methylation alter gene expression or are associated with adverse health effects. PMID:24525453

  2. In vitro effect of cadmium and copper on separated blood leukocytes of Dicentrarchus labrax.

    PubMed

    Vazzana, Mirella; Celi, Monica; Tramati, Cecilia; Ferrantelli, Vincenzo; Arizza, Vincenzo; Parrinello, Nicolň

    2014-04-01

    The immunotoxic effects of heavy metals on blood leukocytes of sea bass (Dicentrarchus labrax) were examined. The cells, separated by a discontinuous Percoll-gradients, were exposed in vitro to various sublethal concentrations of cadmium and copper (10(-7)M, 10(-5)M, and 10(-3)M) and their immunotoxic effect was then evaluated by measuring neutral red uptake, MTT assay, DNA fragmentation and Hsp70 gene expression. First of all, we demonstrated that the cells treated in vitro could incorporate Cd and Cu. A relationship between heavy metal exposure and dose-time-dependent alterations in responses of leukocytes from blood was found for both metals, but copper was more immunotoxic than cadmium in all assays performed. A significant reduction in the cells? ability to uptake neutral red and viability by MTT assay was recorded, indicating that both cadmium and copper could change the membrane permeability, inducing cellular apoptosis when the concentration of metals reached 10(-3)M. The apoptotic effect may also explain the high level of cytotoxicity found when the leukocytes were exposed to higher concentration of metals. These results demonstrated that toxic effect of copper and cadmium affect on the mechanisms of cell-mediated immunity reducing the immune defences of the organism. PMID:24530726

  3. A correlation study of telomere length in peripheral blood leukocytes and kidney function with age.

    PubMed

    Zhang, Wei-Guang; Wang, Yong; Hou, Kai; Jia, Lin-Pei; Ma, Jie; Zhao, De-Long; Zhu, Shu-Ying; Bai, Xiao-Juan; Cai, Guang-Yan; Wang, Yan-Ping; Sun, Xue-Feng; Chen, Xiang-Mei

    2015-06-01

    The current study aimed to investigate the association between telomere length in peripheral blood leukocytes and kidney function in various age groups of a healthy population. A total of 139 healthy individuals were divided into five groups according to their age: 35?44, 45?54, 55?64, 65?74 and >75 years old. Peripheral blood leukocytes were obtained and the telomere restriction fragment (TRF) length was assayed using a digoxigenin?labeled hybridization probe in Southern blot assays. Laboratory assays of kidney function were also performed. A correlation was observed between TRF length and age (r=?0.314, P<0.001), with the telomere length of the individuals >75 years group being significantly shorter than the telomere length of the 35?44, 45?54 and 55?64 years age groups (P<0.05). By contrast, the TRF length for males versus females did not differ for any of the age groups, while a correlation was observed between TRF length and serum levels of cystatin C (r=?0.195, P<0.05). There was also a correlation between TRF length and glomerular filtration rate (r=?0.184, P<0.05). The current study demonstrated that in this cohort, leukocyte telomere length reduced with age and was correlated with serum levels of cystatin C and glomerular filtration rate. Therefore, TRF length is associated with kidney function and may serve as a marker of aging. PMID:25646618

  4. Biogenic amines activate blood leukocytes via trace amine-associated receptors TAAR1 and TAAR2.

    PubMed

    Babusyte, Agne; Kotthoff, Matthias; Fiedler, Julia; Krautwurst, Dietmar

    2013-03-01

    Certain biogenic amines, such as 2-PEA, TYR, or T1AM, modulate blood pressure, cardiac function, brain monoaminergic systems, and olfaction-guided behavior by specifically interacting with members of a group of rhodopsin-like receptors, TAAR. A receptor that is absent from olfactory epithelia but had long been identified in the brain and a variety of peripheral tissues, TAAR1 has been found recently in blood B cells, suggesting a functional role of TAAR1 in these cells. With the present study, we have set out to clarify the expression and functional roles of TAAR in different isolated human blood leukocyte types. Here, we report the functional expression of TAAR1 and its closest relative TAAR2 in blood PMN and T and B cells. Both receptors are coexpressed in a subpopulation of PMN, where they are necessary for the chemosensory migration toward the TAAR1 ligands 2-PEA, TYR, and T1AM, with EC50 values of 0.43 ± 0.05 nM, 0.52 ± 0.05 nM, and 0.25 ± 0.04 nM, respectively. The same amines, with similar potencies, triggered cytokine or Ig secretion, in purified blood T or B cells, respectively. Notably, 2-PEA regulated mRNA expression of 28 T cell function-related genes, above all of the CCL5. In siRNA-guided experiments, TAAR1 and TAAR2 proved to be necessary for amine-induced blood leukocyte functions. In summary, our results demonstrate that biogenic amines potently regulate blood cell functions via TAAR1 and TAAR2 and open the perspective of their specific pharmacological modulation. PMID:23315425

  5. Peripheral blood film review. The demise of the eyecount leukocyte differential.

    PubMed

    Pierre, Robert V

    2002-03-01

    The automated hematology analyzer with CBC and differential results has replaced the traditional manual or individual assay methods for hematologic parameters and the eyecount leukocyte differential as the initial screening and detection system for hematologic abnormalities in modern hospitals and clinics. The traditional review of all automated hematology instrument results by preparation, staining, and microscopic examination of a blood film has disappeared in most institutions. The reasons are the more accurate detection of specimens with distributional or morphologic abnormalities by the instruments than by the traditional eyecount method. The opportunity for a clinician to request a microscopic examination of a blood film, whether or not it is flagged, must be preserved, because the clinician's knowledge of the patient's history, physical findings, and current or prior therapy may indicate review to discover an abnormality that may not have been apparent from the instrument results alone. There has also been a dramatic reduction of the numbers of medical technologists and technicians in medical laboratories. Automation of the CBC and differential counts has reduced the number of technologists needed for performance of these tests. But other factors have had a negative effect, such as the necessity to reduce costs. Consolidation of hematology and chemistry laboratories in core laboratories may produce savings in labor costs, but may also create problems of creating and maintaining areas of expertise, such as hematologic morphology, because of the cross-training required and the necessity of personnel to do all things. This article suggests and documents a number of measures that can be infinity stituted by the laboratory and by clinicians to reduce the number of eyecount differentials and blood film reviews that need to be performed. The first effort is to convince clinicians that valid data exist that confirm that a policy of allowing the laboratory to initiate blood film review based on findings of the CBC and automated differential is a more sensitive and accurate method of detecting patients with blood film abnormalities than routine blood film review of all specimens by technologists. Clinicians need to recognize that daily differential results or differentials at intervals of less than a week are not medically necessary in most patients. The laboratory, however, must provide opportunities for the clinician to request differentials at any time for specific medical reasons. The laboratory must establish the validity of screening criteria for detection of distribution and morphologic abnormalities of leukocytes by clinical correlation studies or adopt criteria established by laboratories with the same instrumentation and which have conducted clinical evaluations. A final observation on the eyecount differential is that it was the only way to identify cell types and their relative proportion for nearly 100 years. Cells were identified by their shape, intracellular structures, and staining characteristics. Many studies were able eventually to correlate some aspect of each cell type's function with their morphologic appearance. It has also been learned that the bone marrow is the source of production of most circulating cells and a great deal of the controls of cell production and release into the peripheral blood have been learned. But leukocytes have many functions, almost none of which are performed in the peripheral blood. The peripheral blood is mainly a conduit from the bone marrow to the tissues where the leukocytes perform their function in the case of the neutrophils and monocytes. It is mainly a recirculation and redistribution system for lymphocytes that usually receive their instructions from antigen processing cells in the tissues and allow these modified cells to home to sites where their functions occur. Cellular morphology and staining characteristics tell little about the maturation stage and functional capabilities of leukocytes. One cannot tell the difference between a band and a segmented neutroph

  6. Modulatory effect of visible light on chemiluminescence of stimulated and nonstimulated blood leukocytes of carp (Cyprinus carpio, L)

    Microsoft Academic Search

    Sandro Belotsky; Ramy R. Avtalion; Harry Friedmann; Rachel Lubart

    1998-01-01

    Irradiation of carp blood leukocytes with a non-laser visible light resulted in a significant inhibition of the spontaneous luminol-dependent chemiluminescence in the cells of a part of the fish. Those leukocytes that were sensitive to the visible light, showed a shorter time-to-peak than the non sensitive, following their stimulation with Ca ionophore. Because a shorter time-to-peak correlates with inflammation, it

  7. Leukocyte esterase urine test

    MedlinePLUS

    Leukocyte esterase is a urine test to look for white blood cells and other signs of infection. ... Leukocyte esterase is a screening test used to detect a substance that suggests there are white blood ...

  8. Effects of radiolabelled monoclonal antibody infusion on blood leukocytes in cancer patients

    SciTech Connect

    Gridley, D.S.; Slater, J.M.; Stickney, D.R. (Loma Linda Univ./Independent Order of Foresters Cancer Research Laboratory, CA (USA))

    1990-01-01

    This study was undertaken to investigate the effects of a single infusion of radiolabelled murine monoclonal antibody (MAb) on peripheral blood leukocytes in cancer patients. Eleven patients with disseminated colon cancer, malignant melanoma, or lung adenocarcinoma were infused with 111In-labelled anti-ZCE 025, anti-p97 type 96.5c, or LA 20207 MAb, respectively. Blood samples were obtained before infusion, immediately after infusion (1 hr), and at 4 and 7 days postinfusion. Flow cytometry analysis of CD3+, CD4+, CD8+, CD16+, and CD19+ lymphocytes showed increasing CD4:CD8 ratios in seven patients after infusion. This phenomenon was not restricted to antibody subclass or to type of cancer. Two of the remaining patients exhibited a marked post-infusion increase in CD8+ cells. In all three patients with malignant melanoma, decreasing levels of CD16+ lymphocytes were noted after infusion and natural killer cell cytotoxicity showed fluctuations which paralleled the changes in the CD16+ subpopulation. Oxygen radical production by phagocytic cells was markedly affected in three subjects. These results suggest that a single infusion of radiolabelled murine MAb may alter the balance of critical lymphocyte subpopulations and modulate other leukocyte responses in cancer patients.

  9. Blood Leukocyte Count on Admission Predicts Cardiovascular Events in Patients with Acute Non-ST Elevation Myocardial Infarction.

    PubMed

    Dharma, Surya; Hapsari, Rosmarini; Siswanto, Bambang B; van der Laarse, Arnoud; Jukema, J Wouter

    2015-06-01

    We aim to test the hypothesis that blood leukocyte count adds prognostic information in patients with acute non-ST-elevation myocardial infarction (non-STEMI). A total of 585 patients with acute non-STEMI (thrombolysis in myocardial infarction risk score???3) were enrolled in this cohort retrospective study. Blood leukocyte count was measured immediately after admission in the emergency department. The composite of death, reinfarction, urgent revascularization, and stroke during hospitalization were defined as the primary end point of the study. The mean age of the patients was 61?±?9.6 years and most of them were male (79%). Using multivariate Cox regression analysis involving seven variables (history of smoking, hypertension, heart rate?>?100 beats/minute, serum creatinine level?>?1.5 mg/dL, blood leukocyte count?>?11,000/µL, use of ?-blocker, and use of angiotensin-converting enzyme inhibitor), leukocyte count?>?11,000/µL demonstrated to be a strong predictor of the primary end point (hazard ratio?=?3.028; 95% confidence interval?=?1.69-5.40, p?blood leukocyte count on admission is an independent predictor of cardiovascular events in patients with acute non-STEMI. PMID:26060384

  10. Direct observation of liposome uptake by leukocytes in vivo in skin blood vessels using intravital fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Devoisselle, Jean-Marie; Mordon, Serge R.; Begu, Sylvie; Desmettre, Thomas

    2000-04-01

    This study aimed to observe liposome uptake by leukocytes in vivo. The study was performed on skin by using a dorsal skin-fold chamber implanted in golden hamsters using intravital microscopy. 5,6-CF-encapsulated PEGylated liposomes were injected intravenously. The skin microcirculation was observed with an intravital Eclipse E800 Nikon microscope fitted with a Xenon light source and an epi-fluorescence assembly. An ultra-high sensitivity video-camera mounted on the microscope projected the image onto a monitor, and the images were recorded for playback analysis with a digital video cassette recorder. An acute inflammatory response was obtained by removing one complete layer of skin and the underlying fascia and avascular tissue on the opposing side of the flap corresponding to an area equivalent to the window aperture. Using these model and set-up, leukocyte rolling and adhesion were easily observed and the entry of PEGylated liposomes into hamster blood leukocytes was studied for a period of 6 hours. PEGylated liposomes were clearly identified alone inside the blood flow and inside the leukocytes as soon as the inflammatory reaction appeared. This study shows for the first time that blood leukocytes in their natural milieu of whole blood are capable of interacting with, and taking up liposomes. This observation is in accordance with previous in vitro studies.

  11. Blood leukocyte subsets and cytokine profile after autologous peripheral blood stem cell transplantation

    Microsoft Academic Search

    S. W. Krause; G. Rothe; M. Gnad; A. Reichle; R. Andreesen

    2003-01-01

    High-dose chemotherapy with autologous peripheral blood stem cell transplantation (PBSCT) includes the risk of infectious complications due to neutropenia and therapy-induced immune deviation. In order to understand early immune recovery in this situation, we analyzed the distribution of cell subsets by flow cytometry and we measured cytokine production in a whole blood assay stimulated with lipopolysaccharide (LPS) in order to

  12. Methods for axolotl blood collection, intravenous injection, and efficient leukocyte isolation from peripheral blood and the regenerating limb.

    PubMed

    Debuque, Ryan J; Godwin, James W

    2015-01-01

    The vertebrate immune system comprises both adaptive and innate immune cells with distinct functions during the resolution of inflammation and wound healing after tissue injury. Recent evidence implicates a requirement for innate immune cells from the myeloid lineage during the early stages of limb regeneration in the Mexican axolotl. Understanding the functions of innate and adaptive immune cells in the axolotl has been hampered by a lack of approaches to isolate and analyze these cells. Here we describe a protocol to isolate myeloid cells from the regenerating axolotl limb that incorporates intravenous delivery of physiological labels. In addition we provide a protocol to enrich for leukocytes in the peripheral blood. These protocols produce single-cell suspensions that can be analyzed using flow cytometry or sorted into specific subsets using fluorescent-activated cell sorting (FACS). FACS is a routine approach to sort cells based on their physical characteristics as well as their cell surface antigen repertoire. Isolated cell populations can then be analyzed in a wide range of downstream assays to facilitate a greater understanding of leukocyte biology in the axolotl. PMID:25740489

  13. Regulation of Interferon-Stimulated Genes in Peripheral Blood Leukocytes in Pregnant and Bred, Nonpregnant Dairy Cows

    Microsoft Academic Search

    C. A. Gifford; K. Racicot; D. S. Clark; K. J. Austin; T. R. Hansen; M. C. Lucy; C. J. Davies; T. L. Ott

    2007-01-01

    In ruminants, pregnancy results in up-regulation of a large number of IFN-stimulated genes (ISG) in the uterus. Recently, one of these genes was also shown to increase in peripheral blood leukocytes (PBL) during early pregnancy in sheep. Our working hypothesis is thatconceptussignalingactivatesmaternalgeneexpres- sion in PBL in dairy cattle. The objectives of this study were to characterize ISG expression in PBL

  14. Receptor Transporter Protein 4 (RTP4) in Endometrium, Ovary, and Peripheral Blood Leukocytes of Pregnant and Cyclic Ewes

    Microsoft Academic Search

    C. A. Gifford; A. M. Assiri; M. C. Satterfield; T. E. Spencer; T. L. Ott

    2008-01-01

    Interferon-tau (IFNT) is secreted by the conceptus trophoblast and signals pregnancy recognition in ruminants. IFNT regulates expression of genes in the endometrium, peripheral blood leukocytes (PBLs), and corpus luteum (CL). Microarray analysis identified that expression of (chemosensory) receptor transport- er protein 4 (RTP4) increased in PBLs during early pregnancy in cows. In the present study, we cloned and characterized RTP4

  15. Expression of IL-17 and COX2 gene in peripheral blood leukocytes of vitiligo patients.

    PubMed

    Esmaeili, Behnaz; Rezaee, Seyed Abdol Rahim; Layegh, Pouran; Tavakkol Afshari, Jalil; Dye, Phil; Ghayoor Karimiani, Ehsan; Kalalinia, Fatemeh; Rafatpanah, Houshang

    2011-06-01

    Vitiligo is a pigmentation disorder in which inflammatory mediators such as cytokines have a pivotal role in disease's pathogenesis. Interleukin 17 (IL-17A) is a proinflammatory cytokine which is involved in the induction of several proinflamatory mediators such as cyclooxygenase 2 (COX2). The aim of this study was to investigate the gene expression of IL-17 and COX2 in peripheral blood leukocytes of vitiligo's patients.Peripheral blood leukocytes from 15 patients with vitiligo and 15 healthy controls were separated using a gradient density centrifugation method. After total RNA isolation and cDNA synthesis, IL-17 and COX2 gene expression were quantified by real-time polymerase chain reaction (PCR). There were no significant differences in IL-17 and COX2 gene expression in lymphocytes of patients with vitiligo compared with control group (p<0.05). However there was an increased IL-17 and COX2 gene expression in neutrophils of patients compared to controls, but it did not reach statistical significance (p=0.05). We could not find any differences in IL-17 and Cox2 gene expression between clinical diseases subtypes, sex and age. There was a significant correlation between IL-17 and COX2 genes expression in the neutrophils of patients (p=0.00, r=0.80). Our results showed an increased expression in IL-17 and Cox-2 genes in neurophils of patients with vitiligo. This suggested that these two factors are involved in the inflammatory process. Further studies with a larger sample size might help to establish the role of these factors in the pathogenesis of diseases. PMID:21625016

  16. Effects of spaceflight on rat peripheral blood leukocytes and bone marrow progenitor cells

    NASA Technical Reports Server (NTRS)

    Ichiki, A. T.; Gibson, L. A.; Jago, T. L.; Strickland, K. M.; Johnson, D. L.; Lange, R. D.; Allebban, Z.

    1996-01-01

    The white blood cell (WBC) elements and the bone marrow myeloid progenitor cell populations were analyzed to ascertain adaptation to micro-gravity and subsequent readaptation to 1 G in rats flown on the 14-day Spacelab Life Sciences-2 (SLS-2) mission. Bone marrow cells were harvested from one group of rats killed inflight (FD13) and blood was drawn from three other groups at various times. The WBC level was normal on FD14 with the exception of neutrophilia. On FD13, numbers of colony-forming units-granulocyte (CFU-G), CFU-GM, and CFU-M from flight animals were decreased compared with ground controls when incubated with recombinant rat interleukin-3 (rrIL-3) alone or in combination with recombinant human erythropoietin (rhEpo). On recovery (R + 0), flight rats had decreased numbers of total leukocytes and absolute numbers of lymphocytes and monocytes with elevated neutrophils compared with control rats. They had lower numbers of CD4, CD8, CD2, CD3, and B cells in the peripheral blood but no differences in spleen lymphocytes.

  17. Gene expression patterns in blood leukocytes discriminate patients with acute infections

    PubMed Central

    Allman, Windy; Chung, Wendy; Mejias, Asuncion; Ardura, Monica; Glaser, Casey; Wittkowski, Knut M.; Piqueras, Bernard; Banchereau, Jacques; Palucka, A. Karolina; Chaussabel, Damien

    2007-01-01

    Each infectious agent represents a unique combination of pathogen-associated molecular patterns that interact with specific pattern-recognition receptors expressed on immune cells. Therefore, we surmised that the blood immune cells of individuals with different infections might bear discriminative transcriptional signatures. Gene expression profiles were obtained for 131 peripheral blood samples from pediatric patients with acute infections caused by influenza A virus, Gram-negative (Escherichia coli) or Gram-positive (Staphylococcus aureus and Streptococcus pneumoniae) bacteria. Thirty-five genes were identified that best discriminate patients with influenza A virus infection from patients with either E coli or S pneumoniae infection. These genes classified with 95% accuracy (35 of 37 samples) an independent set of patients with either influenza A, E coli, or S pneumoniae infection. A different signature discriminated patients with E coli versus S aureus infections with 85% accuracy (34 of 40). Furthermore, distinctive gene expression patterns were observed in patients presenting with respiratory infections of different etiologies. Thus, microarray analyses of patient peripheral blood leukocytes might assist in the differential diagnosis of infectious diseases. PMID:17105821

  18. Effects of 6-formylpterin as an internal source of hydrogen peroxide on cell death of human peripheral blood leukocytes

    Microsoft Academic Search

    Maiko Nishioka; Toshiyuki Arai; Kouhei Yamashita; Masataka Sasada; Hiroko Mori; Hisanari Ishii; Kunihiko Tajima; Keisuke Makino; Kazuhiko Fukuda

    2003-01-01

    The intracellular generation of hydrogen peroxide (H2O2) by 6-formylpterin and its effects on the cell surface exposure of phosphatidylserine (PS) as a marker of cell death were examined in human peripheral blood leukocytes, and the effects were compared with those of exogenously administered H2O2. Neutrophils, monocytes and lymphocytes were isolated from fresh blood, and cultured for 24 h in vitro.

  19. Magnetic Resonance Imaging of Blood Brain/Nerve Barrier Dysfunction and Leukocyte Infiltration: Closely Related or Discordant?

    PubMed Central

    Weise, Gesa; Stoll, Guido

    2012-01-01

    Unlike other organs the nervous system is secluded from the rest of the organism by the blood brain barrier (BBB) or blood nerve barrier (BNB) preventing passive influx of fluids from the circulation. Similarly, leukocyte entry to the nervous system is tightly controlled. Breakdown of these barriers and cellular inflammation are hallmarks of inflammatory as well as ischemic neurological diseases and thus represent potential therapeutic targets. The spatiotemporal relationship between BBB/BNB disruption and leukocyte infiltration has been a matter of debate. We here review contrast-enhanced magnetic resonance imaging (MRI) as a non-invasive tool to depict barrier dysfunction and its relation to macrophage infiltration in the central and peripheral nervous system under pathological conditions. Novel experimental contrast agents like Gadofluorine M (Gf) allow more sensitive assessment of BBB dysfunction than conventional Gadolinium (Gd)-DTPA enhanced MRI. In addition, Gf facilitates visualization of functional and transient alterations of the BBB remote from lesions. Cellular contrast agents such as superparamagnetic iron oxide particles (SPIO) and perfluorocarbons enable assessment of leukocyte (mainly macrophage) infiltration by MR technology. Combined use of these MR contrast agents disclosed that leukocytes can enter the nervous system independent from a disturbance of the BBB, and vice versa, a dysfunctional BBB/BNB by itself is not sufficient to attract inflammatory cells from the circulation. We will illustrate these basic imaging findings in animal models of multiple sclerosis, cerebral ischemia, and traumatic nerve injury and review corresponding findings in patients. PMID:23267343

  20. Angiogenesis by Implantation of Peripheral Blood Mononuclear Cells and Platelets Into Ischemic Limbs

    Microsoft Academic Search

    Osamu Iba; Hiroaki Matsubara; Yoshihisa Nozawa; Soichiro Fujiyama; Katsuya Amano; Yasukiyo Mori; Hiroyuki Kojima; Toshiji Iwasaka

    Background—Peripheral blood mononuclear cells (PBMNCs), platelets, and polymorphonuclear leukocytes (PMNs) contain various angiogenic factors and cytokines. Methods and Results—Unilateral hindlimb ischemia was surgically induced in athymic nude rats, and fluorescence-labeled human blood cells (PBMNCs (107 cells)platelets (109) or PBMNCs (107)platelets (109)PMNs (107)) were intramuscularly implanted into the ischemic limbs. Laser Doppler imaging revealed markedly increased blood perfusion in PBMNCplatelet-implanted limbs

  1. CXCR4 antagonist AMD3100 redistributes leukocytes from primary immune organs to secondary immune organs, lung, and blood in mice.

    PubMed

    Liu, Qian; Li, Zhanzhuo; Gao, Ji-Liang; Wan, Wuzhou; Ganesan, Sundar; McDermott, David H; Murphy, Philip M

    2015-06-01

    AMD3100 (plerixafor), is a specific CXCR4 antagonist approved by the FDA for mobilizing hematopoietic stem cells from bone marrow to blood for transplantation in cancer. AMD3100 also mobilizes most mature leukocyte subsets to blood; however, their source and trafficking potential have not been fully delineated. Here, we show that a single injection of AMD3100 10 mg/kg into C57Bl/6 mice rapidly mobilizes (peak ? 2.5 h) the same leukocyte subsets to blood as in humans. Using this model, we found that AMD3100 mobilization of neutrophils, lymphocytes, and monocytes to blood is not reduced by splenectomy or by blockade of lymphocyte egress from lymph node with FTY720, but is coupled to (i) reduced content of each of these cell types in the bone marrow; (ii) reduced T-cell numbers in thymuses; (iii) increased lymphocytes in lymph nodes; and (iv) increased neutrophil and monocyte content in the lung. Direct intrathymic labeling showed that AMD3100 selectively mobilizes naďve thymic CD4(+) and CD8(+) T cells to blood. Finally, AMD3100-induced neutrophil mobilization to blood did not reduce neutrophil trafficking to thioglycollate-inflamed peritoneum. Thus, AMD3100 redistributes lymphocytes, monocytes, and neutrophils from primary immune organs to secondary immune organs, peripheral tissues, and blood, without compromising neutrophil trafficking to inflamed sites. PMID:25801950

  2. Immunophenotype characteristics of peripheral blood mononuclear leukocytes of chronic idiopathic urticaria patients.

    PubMed

    Garmendia, Jenny V; Zabaleta, Mercedes; Aldrey, Oscar; Rivera, Henry; De Sanctis, Juan B; Bianco, Nicolás E; Blanca, Isaac

    2006-12-01

    The pathogenesis of chronic idiopathic urticaria (CIU) is not completely understood although autoimmunity has been proposed. The aim of the study was to assess the expression of different leukocyte antigens, by flow cytometry, assaying total blood of 29 patients with CIU and of 20 sex and age matched controls. Moreover, we assessed soluble CD154 a marker of immune cell activation, predominantly memory T cells. When patients were divided depending an their response to the autologous serum skin test (ASST), three different groups were encountered: group 1 (n=11): with negative ASST-, group 2 (n=11): positive ASST (ASST+) with normal lymphocyte counts and group 3 (n=7): ASST+ with low lymphocyte counts (< 1500 cells/mm3). A significant increase in CD19+ percentage and not in the absolute count (P < 0.05) was observed in group 1 as compared to controls and to the other groups. In contrast, CD30+, CD45RO+ and CD4+/CD45RO+ percentages and biologically active soluble CD154 levels were significantly higher (P < 0.05) in group 3 as compared to group 1 or to controls. In ASST positive groups, CD45RO+ and CD4+/CD45RO+ positiveness correlates with wheal diameter. In conclusion, memory cells may play a role in these different types of patients and in understanding CIU pathogenesis. PMID:17176904

  3. Effects of resistance exercise and protein ingestion on blood leukocytes and platelets in young and older men

    Microsoft Academic Search

    Juha J. Hulmi; T. Myllymäki; M. Tenhumäki; N. Mutanen; R. Puurtinen; G. Paulsen; A. A. Mero

    2010-01-01

    This study investigated, in a multi-experiment design, the acute effects of milk protein ingestion, aging [50 young (~26 years)\\u000a vs. 45 older (~61 years) men] and training state for the blood leukocyte and platelet responses acutely after a single bout\\u000a of resistance exercise (RE). Moreover, basal effects of 21 weeks of resistance training (RT) were examined. The single bout\\u000a of RE rapidly increased

  4. Flow cytometry as a tool in the evaluation of blood leukocyte function in Chelonia mydas (Linnaeus, 1758) (Testudines, Cheloniidae).

    PubMed

    Rossi, S; Sá-Rocha, V M; Kinoshita, D; Genoy-Puerto, A; Zwarg, T; Werneck, M R; Sá-Rocha, L C; Matushima, E R

    2009-08-01

    Chelonia mydas is a sea turtle that feeds and nests on the Brazilian coast and a disease called fibropapillomatosis is a threat to this species. Because of this, it is extremely necessary to determine a methodology that would enable the analysis of blood leukocyte function in these sea turtles. In order to achieve this aim, blood samples were collected from C. mydas with or without fibropapillomas captured on the Săo Paulo north coast. Blood samples were placed in tubes containing sodium heparin and were transported under refrigeration to the laboratory in sterile RPMI 1640 cell culture medium. Leukocytes were separated by density gradient using Ficoll-PaqueTM Plus, Amershan Biociences. The following stimuli were applied in the assessment of leukocyte function: Phorbol Miristate-Acetate (PMA) for oxidative burst activity evaluation and Zymosan A (Saccharomyces cerevisiae) Bio Particles, Alexa Fluor 594 conjugate for phagocytosis evaluation. Three cell populations were identified: heterophils, monocytes and lymphocytes. Monocytes were the cells responsible for phagocytosis and oxidative burst. PMID:19802451

  5. Effect of gold nanoparticles on production of reactive oxygen species by human peripheral blood leukocytes stimulated with opsonized zymosan.

    PubMed

    Piryazev, A P; Azizova, O A; Aseichev, A V; Dudnik, L B; Sergienko, V I

    2013-11-01

    We studied the effect of gold nanoparticles on ROS production by leukocytes. ROS production was detected by luminol-dependent chemiluminescence (LDCL) of human peripheral blood leukocytes stimulated with opsonized zymosan. Nanoparticle size was 5, 10 and 30 nm. Simultaneous addition of nanoparticles and opsonized zymosan showed that 5-nm nanoparticles inhibited LDCL intensity in comparison with the control, when LDCL recording was conducted in the presence of opsonized zymosan. Increasing nanoparticle size from 5 up to 30 nm enhanced LDCL intensity. Preincubation of gold nanoparticles with autologous blood plasma increased LDCL intensity. In the control (without gold nanoparticles), blood plasma produced no activating effect on LDCL. We found that the effect of gold nanoparticles on leukocyte LDCL depended on nanoparticle size: 10- and 30-nm nanoparticles inhibited LDCL intensity in comparison with the control (incubation in the absence of nanoparticles) irrespective of the duration of incubation, while 5-nm gold nanoparticles had no effect on LDCL intensity. Incubation of gold nanoparticles with autologous plasma increased LDCL intensity if nanoparticle size was 30 and 10 nm. PMID:24319701

  6. Luminol-enhanced chemiluminescence of peripheral blood leukocytes as an early indicator of graft take after allogeneic bone marrow transplantation in patients with acute myelogenous leukaemia

    Microsoft Academic Search

    Allan Rajamäki; Esa-Matti Lilius; Jukka Nikoskelainen; Jaana Proskin; Toivo T. Salmi; Auli Toivanen

    1988-01-01

    The luminol-enhanced chemiluminescence (CL) of peripheral blood leukocytes was studied daily in five patients with acute myelogenous\\u000a leukaemia (AML) in first remission, who were undergoing allogeneic bone marrow transplantation (BMT). The CL was measured\\u000a after stimulation of leukocytes with opsonized zymosan in highly diluted whole blood. All patients had an undetectable CL\\u000a level on day +7, post BMT, simultaneously with

  7. Specific Role of Each Human Leukocyte Type in Viral Infections I. Monocyte as Host Cell for Vesicular Stomatitis Virus Replication In Vitro

    PubMed Central

    Edelman, Robert; Wheelock, E. Frederick

    1967-01-01

    Each major leukocyte type of the peripheral blood of healthy donors was studied in vitro for its ability to support vesicular stomatitis virus (VSV) replication. Purified cultures of each white blood cell type were prepared by the selective adsorption and elution of cells from silicone-treated glass beads. It was found that monocytes and macrophages (derived from the rapid transformation of monocytes in vitro) were the principal host cells for VSV replication. Interferon added to mixed leukocyte cultures, prior to virus inoculation, reduced virus yields and prevented destruction of macrophages. Cultures of small lymphocytes, containing no detectable monocytes or macrophages, produced amounts of virus equivalent to 1% of that produced in leukocyte cultures which contained 7% monocytes. Small lymphocytes did not undergo demonstrable cytopathic alterations in virus-infected cultures. VSV neither replicated nor produced cytopathic effects in polymorphonuclear leukocytes. Images PMID:4316248

  8. Telomere Length in Peripheral Blood Leukocytes Is Associated with Risk of Colorectal Cancer in Chinese Population

    PubMed Central

    Qin, Qin; Sun, Jingwen; Yin, Jieyun; Liu, Li; Chen, Jigui; Zhang, Yuxing; Li, TingTing; Shi, Yun; Wei, Sheng; Nie, Shaofa

    2014-01-01

    Background Human telomeres, tandem repeats of TTAGGG nucleotides at the ends of chromosomes, are essential for maintaining genomic integrity and stability. Results of previous epidemiologic studies about the association of telomere length with risk of colorectal cancer (CRC) have been conflicting. Methods A case-control study was conducted in a Han population in Wuhan, central China. The relative telomere length (RTL) was measured in peripheral blood leukocytes (PBLs) using quantitative real-time polymerase chain reaction (PCR) in 628 CRC cases and 1,256 age and sex frequency matched cancer-free controls. Odds ratios (OR) and 95% confidence intervals (95% CI) were calculated using unconditional logistic regression models to evaluate the association between RTL and CRC risk. Results Using median RTL in the controls as the cutoff, individuals with shorter RTL were associated with a significantly increased risk of CRC (adjusted OR?=?1.27, 95%CI: 1.05–1.55). When participants were further categorized into 3 and 4 groups according to the tertile and quartile RTL values of controls, significant relationships were still observed between shorter RTL and increased CRC risk (OR per tertile?=?1.13, 95%CI: 1.00–1.28, Ptrend?=?0.045; OR per quartile?=?1.12, 95%CI: 1.03–1.23, Ptrend?=?0.012). In stratified analyses, significant association between shorter RTL and increased CRC risk was found in females, individuals younger than 60 years old, never smokers and never drinkers. Conclusions This study suggested that short telomere length in PBLs was significantly associated with an increased risk of CRC in Chinese Han population. Further validation in large prospective studies and investigation of the biologic mechanisms are warranted. PMID:24498432

  9. Inhibition of peripheral blood neutrophil oxidative burst in periodontitis patients with a homeopathic medication Traumeel S

    PubMed Central

    žilinskas, Juozas; žekonis, Jonas; žekonis, Gediminas; Šadzevi?ien?, Renata; Sapragonien?, Marija; Navickait?, Justina; Barzdžiukait?, Ingrida

    2011-01-01

    Summary Background The anti-inflammatory effects of a homeopathic remedy, Traumeel S, have been observed in experimental and clinical studies; however, its antioxidant properties have not been elucidated. The aim of the present study was to evaluate the antioxidant effects of Traumeel S on peripheral blood neutrophils in patients with periodontitis. Material/Methods The study was performed using venous blood of 22 individuals with chronic periodontitis and 21 healthy subjects. The antioxidant effects of Traumeel S on the production of reactive oxygen species by unstimulated and stimulated with unopsonized E. coli neutrophils were investigated using luminol- and lucigenin-dependent chemiluminescence (CL). Results Polymorphonuclear leukocytes of periodontitis patients produced higher levels (p<0.01) of light output of lucigenin-dependent chemiluminescence and significantly reduced (p<0.01) light output of luminol-dependent chemiluminescence than analogous cells of healthy subjects. Highly diluted (10?4 of the stem solution) Traumeel S significantly (by approximately 50%) reduced superoxide-induced oxidation of lucigenin by unstimulated and stimulated with unopsonized E. coli polymorphonuclear leukocytes of periodontitis patients and had a tendency to intensify luminol-dependent chemiluminescence. Preincubation of the unstimulated and stimulated with unopsonized E. coli polymorphonuclear leukocytes of healthy subjects with Traumeel S exerts no inhibitory action on the luminol- and lucigenin-dependent chemiluminescence of the above-mentioned cells. Conclusions This study indicates that Traumeel S may significantly reduce production of superoxide anion by unstimulated and stimulated peripheral blood polymorphonuclear neutrophils of periodontitis patients. PMID:21525811

  10. Change in Peripheral Blood Leukocyte Telomere Length and Mortality in Breast Cancer Survivors

    PubMed Central

    2014-01-01

    Background Progressive telomere shortening with cell division is a hallmark of aging. Short telomeres are associated with increased cancer risk, but there are conflicting reports about telomere length and mortality in breast cancer survivors. Methods We measured peripheral blood leukocyte telomere length at two time points in women enrolled in a multiethnic, prospective cohort of stage I to stage IIIA breast cancer survivors diagnosed between 1995 and 1999 with a median follow-up of 11.2 years. We evaluated associations between telomere length measured at mean 6 (baseline; LTL0; n = 611) and 30 months (LTL30; n = 478) after diagnosis and the change between those time points (n = 478), with breast cancer–specific and all-cause mortality using Cox proportional hazards models adjusted for possible confounders. Statistical tests were two-sided. Results There were 135 deaths, of which 74 were due to breast cancer. Neither baseline nor 30-month telomere length was associated with either all-cause or breast cancer–specific mortality (LTL0: hazard ratio [HR] = 0.83, 95% confidence interval [CI] = 0.67 to 1.02; HR = 0.88; 95% CI = 0.67 to 1.15; LTL30: HR = 0.78, 95% CI = 0.59 to 1.05; HR = 0.86; 95% = CI = 0.58 to 1.26, respectively). However, participants whose telomeres shortened between baseline and 30 months were at a statistically significantly increased risk of breast cancer–specific (HR = 3.03; 95% CI = 1.11 to 8.18) and all-cause mortality (HR = 2.38; 95% CI = 1.28 to 4.39) compared with participants whose telomeres lengthened. When follow-up was censored at 5-years after diagnosis, LTL0 (HR = 0.66; 95% CI = 0.45 to 0.96), LTL30 (HR = 0.51; 95% CI = 0.29 to 0.92), and change in telomere length (HR = 3.45; 95% CI = 1.11 to 10.75) were statistically significantly associated with all-cause mortality. Conclusions Telomere shortening was associated with increased risk of breast cancer–specific and all-cause mortality, suggesting that change in blood telomere length over time could be a biomarker of prognosis. Research on determinants of telomere length and change is needed. PMID:24627273

  11. Maitake beta-glucan promotes recovery of leukocytes and myeloid cell function in peripheral blood from paclitaxel hematotoxicity

    PubMed Central

    Lin, Hong; de Stanchina, Elisa; Zhou, Xi Kathy; Hong, Feng; Seidman, Andrew; Fornier, Monica; Xiao, Wei-Lie; Kennelly, Edward J.; Wesa, Kathleen; Cassileth, Barrie R.

    2011-01-01

    Bone marrow myelotoxicity is a major limitation of chemotherapy. While granulocyte colony stimulating factor (G-CSF) treatment is effective, alternative approaches to support hematopoietic recovery are sought. We previously found that a beta-glucan extract from maitake mushroom Grifola frondosa (MBG) enhanced colony forming unit-granulocyte monocyte (CFU-GM) activity of mouse bone marrow and human hematopoietic progenitor cells (HPC), stimulated G-CSF production and spared HPC from doxorubicin toxicity in vitro. This investigation assessed the effects of MBG on leukocyte recovery and granulocyte/monocyte function in vivo after dose intensive paclitaxel (Ptx) in a normal mouse. After a cumulative dose of Ptx (90–120 mg/kg) given to B6D2F1 mice, daily oral MBG (4 or 6 mg/kg), intravenous G-CSF (80 ?g/kg) or Ptx alone were compared for effects on the dynamics of leukocyte recovery in blood, CFU-GM activity in bone marrow and spleen, and granulocyte/monocyte production of reactive oxygen species (ROS). Leukocyte counts declined less in Ptx + MBG mice compared to Ptx-alone (p = 0.024) or Ptx + G-CSF treatment (p = 0.031). Lymphocyte levels were higher after Ptx + MBG but not Ptx + G-CSF treatment compared to Ptx alone (p < 0.01). MBG increased CFU-GM activity in bone marrow and spleen (p < 0.001, p = 0.002) 2 days after Ptx. After two additional days (Ptx post-day 4), MBG restored granulocyte/monocyte ROS response to normal levels compared to Ptx-alone and increased ROS response compared to Ptx-alone or Ptx + G-CSF (p < 0.01, both). The studies indicate that oral MBG promoted maturation of HPC to become functionally active myeloid cells and enhanced peripheral blood leukocyte recovery after chemotoxic bone marrow injury. PMID:20140432

  12. Subnormal Peripheral Blood Leukocyte Counts Are Related to the Lowest Prevalence and Incidence of Metabolic Syndrome: Tianjin Chronic Low-Grade Systemic Inflammation and Health Cohort Study

    PubMed Central

    Sun, Shaomei; Wu, Hongmei; Zhang, Qing; Wang, Chongjin; Guo, Yinting; Du, Huanmin; Liu, Li; Jia, Qiyu; Wang, Xing; Song, Kun

    2014-01-01

    Few studies have assessed the relationship between a subnormal inflammatory status and metabolic syndrome (MS). We therefore designed a cross-sectional and 5-year cohort study to evaluate how a subnormal peripheral blood leukocyte count is related to MS. Participants were recruited from Tianjin Medical University General Hospital-Health Management Centre. Both a baseline cross-sectional (n = 46,179) and a prospective assessment (n = 13,061) were performed. Participants without a history of MS were followed up for 5 years. Leukocyte counts and MS components were assessed at baseline and yearly during the follow-up. Adjusted logistic and Cox proportional hazards regression models were used to assess relationships between the categories of leukocyte counts and MS. The subnormal leukocyte counts group (1,100–3,900?cells/mm3) had the lowest prevalence and incidence of MS. The odds ratio and hazard ratio (95% confidence interval) of the highest leukocyte counts were 1.98 (1.57–2.49) and 1.50 (1.22–1.84) (both P for trend <0.0001), respectively, when compared to the subnormal leukocyte counts group after adjusting for potential confounders. This study has shown that subnormal leukocyte counts are independently related to the lowest prevalence and incidence of MS. The findings suggest that it is necessary to restudy and discuss the clinical or preventive value of subnormal leukocyte counts. PMID:24876672

  13. White blood cell counts, leukocyte ratios, and eosinophils as inflammatory markers in patients with coronary artery disease.

    PubMed

    Kounis, Nicholas G; Soufras, George D; Tsigkas, Grigorios; Hahalis, George

    2015-03-01

    Inflammation is a key feature of atherosclerosis and its clinical manifestations. The leukocyte count has emerged as a marker of inflammation that is widely available in clinical practice. Since inflammation plays a key role in atherosclerosis and its end results, discovering new biomarkers of inflammation becomes important in order to help diagnostic accuracy and provide prognostic information about coronary cardiac disease. In acute coronary syndromes and percutaneous coronary intervention, elevated levels of almost all subtypes of white blood cell counts, including eosinophils, monocytes, neutrophils, and lymphocytes, and neutrophil-lymphocyte ratio and eosinophil-leukocyte ratio constitute independent predictors of adverse outcomes. Eosinophil count and eosinophil-leukocyte ratio, in particular, emerge as novel biomarkers for risk stratification in patients with coronary artery disease. Since the presence of eosinophils denotes hypersensitivity inflammation and hypersensitivity associated with Kounis syndrome, this reality is essential for elucidating the etiology of inflammation in order to consider predictive and preventive measures and to apply the appropriate therapeutic methods. PMID:24770327

  14. [The study of aberrant methylation in blood leukocytes of liquidators of the Chernobyl accident].

    PubMed

    Kuz'mina, N S; Miazin, A E; Lapteva, N Sh; Rubanovich, A V

    2014-01-01

    The study of aberrant methylation of CpG islands in the promoter regions of genes (P16/CDKN2A, P14/ARF, RASSF1A, GSTP1) in blood leukocytes of liquidators of the Chernobyl accident (n = 83, 38-76 years of age) and control subjects of two groups (n = 48, age ? 35 and n = 65, age > 35) was carried out using methylation-sensitive restriction endonuclease analysis followed by PCR. The total number of AciI sites in the analyzed fragments ranged from 2 to 7 for different genes. Only 1 subject (2.1%) from the control group (healthy young individuals, age ? 35) has methylation of the studied CpG--dinucleotides of RASSF1A gene. Promoter methylation of at least one of the genes analyzed was observed in 28.92% liquidators and significantly exceeded (p = 0.016) such rate in a one-age (> 35 years of age) control group (12.31%). A significantly elevated frequency (p = 0.023) of individuals with abnormal methylation of GSTP1 gene in the group of liquidators as compared to the control group was revealed. The occurrence of promoter methylation of RASSF1A gene significantly correlated with aging both in the control group (r = 0.214; p = 0.023) and in the liquidators of the Chernobyl accident (r = 0.230; p = 0.036). No similar trend was found for other genes. Multiple regression analysis showed that the growth in the number of methylated loci of a set of genes p16, p14 and GSTP1 is exclusively due to the fact of exposure (OR = 7.32, 95% CI = 2.49-25.83, p-value = 2.7 x 10(-5)). The results obtained demonstrate for the first time the reality of the radiation-induced aberrant methylation of CpG islands in promoters of genes involved in the basic protective, functions of cells in the human body in remote periods after radiation exposure. PMID:25764814

  15. Bacteria and zymosan opsonized with histone, dextran sulfate, and polyanetholesulfonate trigger intense chemiluminescence in human blood leukocytes and platelets and in mouse macrophages

    Microsoft Academic Search

    Isaac Ginsburg; Ruth Borinsky; Meir Lahav; K. E. Gillert; Sabina Falkenberg; Michael Winkler; Sybille Muller

    1982-01-01

    Human blood leukocytes and platelets and mouse peritoneal macrophages emit very rapid and very intense Luminol-dependent chemiluminescence (CL) signals when treated with streptococci, staphylococci, or with zymosan, which have been preopsonized with arginine-rich histone, dextran sulfate or polyanetholesulfonate (liquoid). Liquoid alone at 10–30µg\\/2×105 leukocytes also triggers intense CL responses in the absence of a carrier. Strong CL can also be

  16. Nadolol inhibits reactive oxygen species generation by leukocytes and linoleic acid oxidation

    Microsoft Academic Search

    Cesar H Magsino; Wael Hamouda; Vini Bapna; Husam Ghanim; Ihab A Abu-Reish; Ahmad Aljada; Paresh Dandona

    2000-01-01

    We studied the effect of short-term nadolol administration on the reactive oxygen species (ROS) generation by polymorphonuclear leukocytes and mononuclear cells in 8 normal subjects. At a oral dose of 40 mg\\/day for 5 days, nadolol produced a decrease in the ROS generation by leukocytes. ROS generation by polymorphonuclear leukocytes decreased by 38% from 134 ± 44 mV at baseline

  17. Repetitive element hypomethylation in blood leukocyte DNA and cancer incidence, prevalence, and mortality in elderly individuals: the Normative Aging Study

    Microsoft Academic Search

    Zhong-Zheng Zhu; David Sparrow; Lifang Hou; Letizia Tarantini; Valentina Bollati; Augusto A. Litonjua; Antonella Zanobetti; Pantel Vokonas; Robert O. Wright; Andrea Baccarelli; Joel Schwartz

    2011-01-01

    Background  Global genomic hypomethylation is a common epigenetic event in cancer that mostly results from hypomethylation of repetitive\\u000a DNA elements. Case–control studies have associated blood leukocyte DNA hypomethylation with several cancers. Because samples\\u000a in case–control studies are collected after disease development, whether DNA hypomethylation is causal or just associated\\u000a with cancer development is still unclear.\\u000a \\u000a \\u000a \\u000a \\u000a Methods  In 722 elderly subjects from the

  18. Chemiluminescence response of whole blood in patients undergoing urological operations

    Microsoft Academic Search

    M. Sakumoto; T. Matsumoto; O. Mochida; S. Kubo; Y. Mizunoe; J. Kumazawa

    1997-01-01

    Polymorphonuclear leukocytes (PMNs) are one of the most important components of the defence mechanisms against bacterial infection.\\u000a The functions of PMNs are believed to be impaired in patients during the perioperative period. Bactericidal function of PMNs\\u000a was investigated together with the luminol-dependent chemiluminescence (CL) reaction of whole blood in 23 patients, 12 undergoing\\u000a open surgery and 11 undergoing endoscopic surgery.

  19. Proficiency Testing of Human Leukocyte Antigen-DR and Human Leukocyte Antigen-DQ Genetic Risk Assessment for Type 1 Diabetes Using Dried Blood Spots

    PubMed Central

    Dantonio, Paul; Meredith-Molloy, Nancy; Hagopian, William A.; She, Jin Xiong; Akolkar, Beena; Cordovado, Suzanne K.; Hendrix, Miyono; Henderson, L. Omar; Hannon, W. Harry; Vogt, Robert F.

    2010-01-01

    Background The plurality of genetic risk for developing type 1 diabetes mellitus (T1DM) lies within the genes that code for the human leukocyte antigens (HLAs). Many T1DM studies use HLA genetic risk assessment to identify higher risk individuals, and they often conduct these tests on dried blood spots (DBSs) like those used for newborn bloodspot screening. One such study is The Environmental Determinants of Diabetes in the Young (TEDDY), a long-term prospective study of environmental risk factors. To provide quality assurance for T1DM studies that employ HLA genetic risk assessment, the Centers for Disease Control and Prevention (CDC) conducts both a voluntary quarterly proficiency testing (VQPT) program available to any laboratory and a mandatory annual proficiency testing (PT) challenge for TEDDY laboratories. Methods Whole blood and DBS samples with a wide range of validated HLA-DR and HLA-DQ genotypes were sent to the participating laboratories. Results were evaluated on the basis of both the reported haplotypes and the HLA genetic risk assessment. Results Of the reported results from 24 panels sent out over six years in the VQPT, 94.7% (857/905) were correctly identified with respect to the relevant HLA-DR or HLA-DQ alleles, and 96.4% (241/250) were correctly categorized for risk assessment. Significant improvement was seen over the duration of this program, usually reaching 100% correct categorization during the last three years. Of 1154 reported results in four TEDDY PT challenges, 1153 (99.9%) were correctly identified for TEDDY eligibility. Conclusions The different analytical methods used by T1DM research centers all provided accurate (>99%) results for genetic risk assessment. The two CDC PT programs documented the validity of the various approaches to screening and contributed to overall quality assurance. PMID:20663459

  20. [The role of blood plasma porphyrins in the effects of He-Ne-laser on human leukocytes].

    PubMed

    Klebanov, G I; Chichuk, T V; Osipov, A N; Vladimirov, Iu A

    2005-01-01

    The He-Ne-laser induced effects in human blood leukocytes in the presence of autologic plasma were investigated. Experiments were performed in two ways: (1) He-Ne-laser irradiation of cells in the presence of autologic plasma or (2) laser irradiation and subsequent addition of autologic plasma to the cell suspension. The concentration dependencies of plasma additions were evaluated. To obtain different concentrations of porphyrins in plasma samples, we either diluted the samples with PBS or selected patients with different porphyrin plasma content. The effects of He-Ne-laser irradiation were characterized by the maximum effect dose (Dmax) of irradiation and the degree of maximum cell activation (Amax, priming index). In the first series of experiments, we irradiated leukocytes in the presence of autologic plasma taken from patients with pneumonia and bronchial asthma. It was found that Dmax decreased with increasing porphyrin concentration in plasma. It was observed that, at low porphyrin concentrations, Amax increased severalfold with increasing photosensitizer concentration. At a porphyrin concentration of 0.46 pmol a decrease in Amax was detected as the porhyrin concentration increased. The same effects were revealed at high doses of laser irradiation. Very similar effects were found in experiments with the addition of irradiated plasma to cells. However, the Amax value was considerably less compared to that after irradiation in the presence of plasma (160% vs. 230 - 270% upon combined irradiation of cells and plasma). The Dmax value was higher in the series of experiments in which plasma was irradiated separately. The results suggest that laser-induced leukocyte activation can be mediated by blood plasma porphyrins and the products of lipid peroxidation formed as a result of porphyrin-photosensitized lipid oxidation. PMID:16212065

  1. Secretory Leukocyte Protease Inhibitor: A Macrophage Product Induced by and Antagonistic to Bacterial Lipopolysaccharide

    Microsoft Academic Search

    Fen-yu Jin; Carl Nathan; Danuta Radzioch; Aihao Ding

    1997-01-01

    To explore regulation of potentially lethal responses to bacterial lipopolysaccharide (LPS), we used differential display under LPS-free conditions to compare macrophage cell lines from two strains of mice congenic for a locus affecting LPS sensitivity. LPS- hyporesponsive cells, primary macrophages, and polymorphonuclear leukocytes transcribed secretory leukocyte protease inhibitor (SLPI), a known epithelial cell-derived inhibitor of leukocyte serine proteases. Transfection of

  2. Leukocyte Recruitment of Airways by Cigarette Smoke and Particle Phase in Contrast to Cytotoxicity of Vapor

    Microsoft Academic Search

    Kaye H. Kilburn; Wayland McKenzie

    1975-01-01

    After hamsters had breathed fresh cigarette smoke in a miniature chamber, airways of the lung showed recruitment of polymorphonuclear leukocytes. Exposure to particles alone by removal of the vapor phase with charcoal did not change the leukocyte response. However, exposure to cigarette smoke vapor after removal of particles with Cambridge filters did not recruit leukocytes but produced nuclear and cytoplasmic

  3. Effect of two-chambered bicarbonate lactate-buffered peritoneal dialysis fluids on peripheral blood mononuclear cell and polymorphonuclear cell function in vitro.

    PubMed

    Sundaram, S; Cendoroglo, M; Cooker, L A; Jaber, B L; Faict, D; Holmes, C J; Pereira, B J

    1997-11-01

    Low pH, high osmolality, increasing glucose concentration, and glucose degradation products (GDP) formed during heat sterilization of conventional peritoneal dialysis (PD) fluids have been shown to have a detrimental effect on cells involved in peritoneal host defense. The two-chambered PD fluid bag in which glucose at pH approximately 3 is separated from a bicarbonate (25 mmol/L)-lactate (15 mmol/L) buffer during heat sterilization permits PD fluids with lower GDP to be delivered to the patient at neutral pH. To establish the possible benefit of two-chambered bag PD fluids on peripheral blood mononuclear cell (PBMC) and polymorphonuclear (PMN) cell function, we compared conventional 1.5% Dianeal (1.5%D) with 1.5% two-chambered bag bicarbonate-lactate (1.5%D-B), and conventional 4.25% Dianeal (4.25%D) with 4.25% two-chambered bag bicarbonate-lactate (4.25%D-B). Furthermore, to study the effect of the sterilization process on PBMC and PMN function, we compared filter-sterilized 4.25%D (4.25%D-F) with 4.25%D and 4.25%D-B. PBMC were harvested by Ficoll-Hypaque separation, and 2.5 x 10(6) cells in RPMI were incubated with an equal volume of the test fluids for 4 hours, pelleted, and resuspended in RPMI containing 10 ng endotoxin for a further 20 hours. Tumor necrosis factor alpha (TNF-alpha) production by endotoxin-stimulated PBMC was not significantly different (P = 0.10) between 1.5%D-B and 1.5%D, but was significantly higher (P = 0.01) with 4.25%D-B compared with 4.25%D. PBMC exposed to filter-sterilized fluid (4.25%D-F) showed significantly higher endotoxin-stimulated TNF-alpha production compared with 4.25%D (P = 0.02), but was not significantly different from 4.25%D-B (P = 0.40). PMN were harvested by Ficoll-Hypaque separation and 10 x 10(6) cells incubated with test fluids for 30 minutes. After incubation, phagocytosis (phagocytosis index) was determined by the uptake of 14C-labeled Staphylococcus aureus, oxidative burst by reduction of ferricytochrome C to ferrocytochrome C on stimulation with PMA, and enzyme release by measurement of endotoxin-stimulated bactericidal/permeability increasing protein (BPI). Bicarbonate-lactate two-chambered fluids of similar osmolality and glucose concentration conferred a significant improvement in phagocytosis (P = 0.02 for 1.5%D-B and P < 0.001 for 4.25%D-B). Oxidative burst and BPI release were significantly higher in 4.25%D-B compared with 4.25%D (P < 0.001). Filter-sterilized 4.25%D-F conferred a significant improvement in phagocytosis and oxidative burst compared with 4.25%D (P < 0.001) or 4.25%D-B (P < 0.001). Furthermore, conventional 4.25%D was associated with significantly lower BPI release compared with 4.25%D-F (P = 0.01). GDP's acetaldehyde and 5-HMF were analyzed in 4.25%D-B, 4.25%D, and 4.25%D-F. Acetaldehyde was below the lower limit (0.79 ppm) of the standard curve in 4.25%D-B and 4.25%D-F fluids but was detected (3.76 to 5.12 ppm) in all of the 4.25%D fluids. Relative levels of 5-HMF in the 4.25%D-B (0.032 to 0.041 Abs @ 284 nm) and 4.25%D (0.031 to 0.036 Abs @ 284 nm) were similar. The lowest levels (0.001 Abs @ 284 nm) were observed in the filter-sterilized 4.25%D-F. The beneficial effects of two-chambered bicarbonate lactate-buffered PD fluids on PBMC and PMN function are probably related to reduction of GDP from heat sterilization of glucose in a separate chamber at a lower pH. This improvement in biocompatibility could have a beneficial affect on peritoneal defenses. PMID:9370184

  4. The effects of oil exposure on peripheral blood leukocytes and splenic melano-macrophage centers of Gulf of Mexico fishes.

    PubMed

    Ali, Ahmad Omar; Hohn, Claudia; Allen, Peter J; Ford, Lorelei; Dail, Mary Beth; Pruett, Stephen; Petrie-Hanson, Lora

    2014-02-15

    In August and November 2010 we collected and examined peripheral blood and tissues from three species of Gulf of Mexico fish. Findings were compared to non-exposed control fish. The leukocyte counts of exposed alligator gar were not significantly different from controls, while exposed Gulf killifish and sea trout had significantly decreased lymphocyte counts. Liver ethoxyresorufin-O-deethylase (EROD) values from sea trout were significantly greater than control sea trout EROD values, suggesting poly aromatic hydrocarbon exposure. Splenic melano-macrophage centers (MMCs) from exposed sea trout and Gulf killifish showed a significant increase in number compared to non-exposed fish. Sea trout splenic MMCs were also significantly greater in size. These findings suggest that Gulf fish sampled were exposed to crude oil from the Macondo well and were in a lymphopenic or immuno-compromised state. PMID:24405733

  5. Expression of Adiponectin Receptors on Peripheral Blood Leukocytes of Hypertensive Children Is Associated with the Severity of Hypertension

    PubMed Central

    Gackowska, Lidia; Litwin, Mieczyslaw; Trojanek, Joanna; Eljaszewicz, Andrzej; Kubiszewska, Izabela; Niemirska, Anna; Wierzbicka, Aldona; Michalkiewicz, Jacek

    2015-01-01

    The aim of the study was to find out whether peripheral blood leukocyte adiponectin receptors 1 and 2 (AdipoR1, AdipoR2) protein expression patterns (flow cytometry) differ between the primary hypertension children (n = 57) and healthy controls (n = 19) and if their expression levels are related to selected clinical parameters. The group of 26 patients [AdipoR(?)] showed lower and the group of 31 patients [AdipoR(+)] showed higher AdipoRs protein expression than the control and each other (P < 0.01 for neutrophils, P < 0.05 for monocytes). The AdipoR(+) leukocytes expressed higher AdipoR1 mRNA levels (RT-PCR) than AdipoR(?) ones and controls (P = 0.022 and P = 0.007, resp.). Despite greater BMI, the AdipoR(?) patients had unchanged serum adiponectin levels. In contrast, AdipoR(+) patients had lower serum adiponectin concentrations than the AdipoR(?) ones and controls (P < 0.001). The AdipoR(+) patients had higher blood pressure (P = 0.042) and greater carotid intima-media thickness (P = 0.017) than the AdipoR(?) ones. The stage of hypertension was associated with increased neutrophil but not monocyte AdipoR1 density (AdipoR1 MFI) (P < 0.05). Severe ambulatory hypertension was presented more often in AdipoR(+) patients than in AdipoR(?) ones (51.6% versus 26.9%, resp.; P < 0.01). In conclusion, neutrophil AdipoRs upregulation was associated with early stages of vascular injury, hypertension severity, and low serum levels of adiponectin. PMID:26146630

  6. Increased expression of peripheral blood leukocyte genes implicate CD14+ tissue macrophages in cellular intestine allograft rejection.

    PubMed

    Ashokkumar, Chethan; Ningappa, Mylarappa; Ranganathan, Sarangarajan; Higgs, Brandon W; Sun, Qing; Schmitt, Lori; Snyder, Sara; Dobberstein, Jennifer; Branca, Maria; Jaffe, Ronald; Zeevi, Adriana; Squires, Robert; Alissa, Feras; Shneider, Benjamin; Soltys, Kyle; Bond, Geoffrey; Abu-Elmagd, Kareem; Humar, Abhinav; Mazariegos, George; Hakonarson, Hakon; Sindhi, Rakesh

    2011-10-01

    Recurrent rejection shortens graft survival after intestinal transplantation (ITx) in children, most of whom also experience early acute cellular rejection (rejectors). To elucidate mechanisms common to early and recurrent rejection, we used a test cohort of 20 recipients to test the hypothesis that candidate peripheral blood leukocyte genes that trigger rejection episodes would be evident late after ITx during quiescent periods in genome-wide gene expression analysis and would achieve quantitative real-time PCR replication pre-ITx (another quiescent period) and in the early post-ITx period during first rejection episodes. Eight genes were significantly up-regulated among rejectors in the late post-ITx and pre-ITx periods, compared with nonrejectors: TBX21, CCL5, GNLY, SLAMF7, TGFBR3, NKG7, SYNE1, and GK5. Only CCL5 was also up-regulated in the early post-ITx period. Among resting peripheral blood leukocyte subsets in randomly sampled nonrejectors, CD14(+) monocytes expressed the CCL5 protein maximally. Compared with nonrejectors, rejectors demonstrated higher counts of both circulating CCL5(+)CD14(+) monocytes and intragraft CD14(+) monocyte-derived macrophages in immunohistochemistry of postperfusion and early post-ITx biopsies from the test and an independent replication cohort. Donor-specific alloreactivity measured with CD154(+) T-cytotoxic memory cells correlated with the CCL5 gene and intragraft CD14(+) monocyte-derived macrophages at graft reperfusion and early post-ITx. CCL5 gene up-regulation and CD14(+) macrophages likely prime cellular ITx rejection. Infiltration of reperfused intestine allografts with CD14(+) macrophages may predict rejection events. PMID:21854741

  7. A simple method for preparing neocyte-enriched leukocyte-poor blood for transfusion-dependent patients.

    PubMed

    Hogan, V A; Blanchette, V S; Rock, G

    1986-01-01

    Recent treatment for patients with thalassemia and chronic anemia involves transfusion of young red cells (YRBCs) or "neocytes." We developed a technique enabling YRBCs to be separated based on their buoyant density in autologous plasma during centrifugation. Following this procedure, measurement of pyruvate kinase (PK), an age-dependent red cell enzyme, showed neocyte enrichment in the top one-third of the RBC layer corresponding to a mean of 47.5 percent of the total PK present in the unfractionated unit. To provide a neocyte transfusion preparation with an acceptable hemoglobin content, the top one-third fraction from each of three bags of blood was pooled. Leukocytes were removed from this "neocyte unit" by an initial sedimentation with 6 percent hydroxyethyl starch (HES) followed by filtration through a filter (IG 500, Imugard). This process resulted in removal of 99.3 +/- 0.5 percent (mean +/- SD) of the leukocytes with a mean RBC recovery of 89.5 +/- 5.5 percent and a final hemoglobin content of 53.4 +/- 2.3 g. Tests for plasma-free hemoglobin and HES in the supernatant of the final transfusion product gave acceptable mean values of 28 mg per dl and 3.0 mg per ml. Autologous mean RBC survival of Cr51-labeled YRBC fractions was 41.8 +/- 2.9 days (n = 5). This technique yields neocyte enrichment superior to that achieved using a cell processor (model 2991, IBM) and has the added advantage of being less costly to prepare ($45.00 [1984] U.S. per YRBC unit as compared to an estimated $135.00 [1984] U.S., IBM) and more economical in terms of blood use. PMID:3705144

  8. Predictive Levels of CD24 in Peripheral Blood Leukocytes for the Early Detection of Colorectal Adenomas and Adenocarcinomas

    PubMed Central

    Kraus, Sarah; Shapira, Shiran; Kazanov, Dina; Naumov, Inna; Moshkowitz, Menachem; Santo, Erwin; Galazan, Lior; Shmueli, Einat; Hallack, Aharon; Arber, Nadir

    2015-01-01

    CD24 is expressed in 90% of colorectal adenomas and adenocarcinomas. Colorectal cancer (CRC) can be mostly prevented but average risk population screening by stool testing or colonoscopy faces many hurdles. Blood testing is clinically needed. We aimed to evaluate the utility of CD24 expression in peripheral blood leukocytes (PBLs). Two independent case studies were conducted in eligible individuals undergoing colonoscopy. Protein extracted from PBLs was subjected to immunoblotting using anti-CD24 monoclonal antibodies. CD24 sensitivity and specificity were determined using receiver operating characteristic (ROC) analysis. Initially, 150 subjects were examined: 63 had CRC, 19 had adenomas, and 68 had normal colonoscopies. The sensitivity and specificity of CD24 for distinguishing CRC from normal subjects were 70.5% (95% CI, 54.8–83.2%) and 83.8% (95% CI, 74.6–92.7%) and for adenomas 84.2% (95% CI, 60.4–96.4%) and 73.5% (95% CI, 61.4–83.5%), respectively. In the second trial (n = 149), a similar specificity but higher sensitivity was achieved: 80.0% (95% CI, 63.1–91.6%) for CRC and 89.2% (95% CI, 74.6–97%) for adenomas. A simple noninvasive blood test evaluating CD24 levels has high sensitivity and specificity for detecting colorectal adenomas and cancer in patients undergoing colonoscopy at an urban medical center. Larger multicenter studies are warranted to establish the potential of this promising test.

  9. Hydrodynamic Shear Rate Regulates Melanoma-Leukocyte Aggregation, Melanoma Adhesion to the Endothelium, and Subsequent Extravasation

    E-print Network

    Dong, Cheng

    Hydrodynamic Shear Rate Regulates Melanoma-Leukocyte Aggregation, Melanoma Adhesion that polymorphonuclear neutrophils (PMNs) may enhance melanoma adhesion to the endothelium (EC) and subsequent microenvironment within the microcirculation. In this study, effects of hydrodynamic flow on regulating melanoma

  10. Hydrodynamic Shear Rate Regulates Melanoma-Leukocyte Aggregation, Melanoma Adhesion to the Endothelium, and

    E-print Network

    Simon, Scott I.

    Hydrodynamic Shear Rate Regulates Melanoma-Leukocyte Aggregation, Melanoma Adhesion that polymorphonuclear neutrophils (PMNs) may enhance melanoma adhesion to the endothelium (EC) and subsequent microenvironment within the microcirculation. In this study, effects of hydrodynamic flow on regulating melanoma

  11. Recombinant Gamma Interferon Increases the Binding of Peripheral Blood Mononuclear Leukocytes and a Leu3+ T Lymphocyte Clone to Cultured Keratinocytes and to a Malignant Cutaneous Squamous Carcinoma Cell Line That Is Blocked by Antibody Against the LFA-1 Molecule

    Microsoft Academic Search

    Brian J. Nickoloff; David M. Lewinsohn; Eugene C. Butcher; Alan M. Krensky; Carol Clayberger

    1988-01-01

    Because keratinocytes (KCs) express HLA-DR in a wide variety of skin diseases in which mononuclear leukocytes are observed in close apposition to KCs (i.e., graft-versus-host disease), and since gamma interferon (IFN-?) induces HLA- DR expression on KCs, we asked whether IFN-? treatment of KCs would influence the adherence of mononuclear leukocytes. When allogeneic peripheral blood mononuclear leukocytes (PBML) and a

  12. Effect of oxidatively modified and non-modified human serum albumin on luminol-dependent chemiluminescence of human peripheral blood leukocytes stimulated with opsonized zymosan.

    PubMed

    Piryazev, A P; Azizova, A P; Aseichev, A V; Sergienko, V I

    2014-07-01

    We studied the effects of native and oxidized human serum albumin on luminol-dependent chemiluminescence of human peripheral blood leukocytes stimulated with opsonized zymosan. Human serum albumin was added simultaneously with opsonized zymosan at the beginning of the chemiluminescent reaction. Otherwise, leukocytes were incubated with human serum albumin at 37°C for various periods before addition of opsonized zymosan. Oxidized human serum albumin was obtained by the method of metal-catalyzed oxidation. In control to non-modified albumin, oxidized albumin produced an inhibitory effect on luminol-dependent chemiluminescence of leukocytes. These changes were observed in experiments with addition of oxidized albumin at the beginning of a chemiluminescent reaction and after incubation of study agent with cells. PMID:25065314

  13. IL-6 blockade reverses the abnormal STAT activation of peripheral blood leukocytes from rheumatoid arthritis patients.

    PubMed

    Ortiz, M A; Diaz-Torné, C; Hernández, M V; Reina, D; de la Fuente, D; Castellví, I; Moya, P; Ruiz, J M; Corominas, H; Zamora, C; Cantó, E; Sanmartí, R; Juarez, C; Vidal, S

    2015-06-01

    Considering the interplay of multiple STATs in response to cytokines, we investigated how IL-6 and its blocking affect STAT signaling in rheumatoid arthritis (RA). Leukocytes obtained from RA patients before and after tocilizumab treatment and healthy donors (HDs) were cytokine-stimulated and STAT phosphorylation was analyzed by cytometry. RA patients had significantly fewer pSTAT1+, pSTAT3+, and pSTAT6+ monocytes and pSTAT5+ lymphocytes than HDs. After 24weeks of treatment, percentages of IFN?-induced pSTAT1+ and IL-10-induced pSTAT3+ monocytes in RA patients increased, reaching levels comparable to HDs. pSTAT1+ and pSTAT3+ cells correlated inversely with RA disease activity index and levels of pSTAT+ cells at baseline were higher in patients with good EULAR response to tocilizumab. IFN?-induced pSTAT1+ cells correlated inversely with memory T cells and anti-CCP levels. IL-10-induced pSTAT3+ cells correlated with Treg/Teff ratio. Our findings suggest that IL-6 blocking reduces the inflammatory mechanisms through the correction of STAT1 and STAT3 activation status. PMID:25847223

  14. Periodic oscillation of blood leukocytes, platelets, and hemoglobin in a patient with chronic eosinophilic leukemia.

    PubMed

    Xiao, Zhijian; Hao, Yushu; Qin, Tiejun; Han, Zhongchao

    2003-01-01

    Chronic eosinophilic leukemia (CEL) is a rare myeloproliferative disease in which autonomous, clonal proliferation of eosinophilic precursors results in persistent increase of eosinophils in the blood and bone marrow. A case of CEL spontaneous oscillation of white blood cell (WBC) count is presented. The cycle of WBC variation comprised about 60 days. Similar cyclic variations were noted in his platelet count, hemoglobin level and bone marrow cellularity, as well as in the spleen size, which was directly correlated with the WBC count. The numbers of bone marrow erythroid colony-forming units (CFU-E), granulocyte-macrophage colony-forming units (CFU-GM) and the serum level of colony-stimulating factors (CSFs) were also regularly changed during the oscillation of WBC. Bone marrow hyperplasia was accompanied with the increase in peripheral WBC count, suggesting that the variation of cell production caused the cyclic oscillation. PMID:12479858

  15. An extended convection diffusion model for red blood cell-enhanced transport of thrombocytes and leukocytes

    NASA Astrophysics Data System (ADS)

    Hund, S. J.; Antaki, J. F.

    2009-10-01

    Transport phenomena of platelets and white blood cells (WBCs) are fundamental to the processes of vascular disease and thrombosis. Unfortunately, the dilute volume occupied by these cells is not amenable to fluid-continuum modeling, and yet the cell count is large enough that modeling each individual cell is impractical for most applications. The most feasible option is to treat them as dilute species governed by convection and diffusion; however, this is further complicated by the role of the red blood cell (RBC) phase on the transport of these cells. We therefore propose an extended convection-diffusion (ECD) model based on the diffusive balance of a fictitious field potential, ?, that accounts for the gradients of both the dilute phase and the local hematocrit. The ECD model was applied to the flow of blood in a tube and between parallel plates in which a profile for the RBC concentration field was imposed and the resulting platelet concentration field predicted. Compared to prevailing enhanced-diffusion models that dispersed the platelet concentration field, the ECD model was able to simulate a near-wall platelet excess, as observed experimentally. The extension of the ECD model depends only on the ability to prescribe the hematocrit distribution, and therefore may be applied to a wide variety of geometries to investigate platelet-mediated vascular disease and device-related thrombosis.

  16. A dolphin peripheral blood leukocyte cDNA microarray for studies of immune function and stress reactions.

    PubMed

    Mancia, Annalaura; Lundqvist, Mats L; Romano, Tracy A; Peden-Adams, Margie M; Fair, Patricia A; Kindy, Mark S; Ellis, Blake C; Gattoni-Celli, Sebastiano; McKillen, David J; Trent, Harold F; Chen, Yian Ann; Almeida, Jonas S; Gross, Paul S; Chapman, Robert W; Warr, Gregory W

    2007-01-01

    A microarray focused on stress response and immune function genes of the bottlenosed dolphin has been developed. Random expressed sequence tags (ESTs) were isolated and sequenced from two dolphin peripheral blood leukocyte (PBL) cDNA libraries biased towards T- and B-cell gene expression by stimulation with IL-2 and LPS, respectively. A total of 2784 clones were sequenced and contig analysis yielded 1343 unigenes (archived and annotated at ). In addition, 52 dolphin genes known to be important in innate and adaptive immune function and stress responses of terrestrial mammals were specifically targeted, cloned and added to the unigene collection. The set of dolphin sequences printed on a cDNA microarray comprised the 1343 unigenes, the 52 targeted genes and 2305 randomly selected (but unsequenced) EST clones. This set was printed in duplicate spots, side by side, and in two replicates per slide, such that the total number of features per microarray slide was 19,200, including controls. The dolphin arrays were validated and transcriptomic profiles were generated using PBL from a wild dolphin, a captive dolphin and dolphin skin cells. The results demonstrate that the array is a reproducible and informative tool for assessing differential gene expression in dolphin PBL and in other tissues. PMID:17084893

  17. Effect of oral administration of kynurenic acid on the activity of the peripheral blood leukocytes in mice

    PubMed Central

    Siwicki, Andrzej Krzysztof; Wójcik, Roman Marcin; Kaczorek, Edyta; Turski, Waldemar Andrzej

    2014-01-01

    Kynurenic acid (KYNA), an endogenous tryptophan metabolite, is a selective ligand of the GPR35 receptor, expressed mainly on the immune cells. In inflammatory conditions, by affecting this receptor, KYNA inhibits the synthesis of pro-inflammatory cytokines and probably protects tissues from oxidative damage. However, we lack data regarding the effect of exogenous KYNA on the activity of immune cells in healthy individuals. The objective of this study has been to determine the influence of kynurenic acid administered to mice in different doses (2.5, 25 or 250 mg/l) and for different time periods (3, 7, 14, 28 days) in drinking water, on the activity of their peripheral blood leukocytes. The determinations comprised the proliferative activity of lymphocytes (MTT assay) and the phagocytic activity as well as the respiratory burst activity of granulocytes and monocytes (Phagotest, Phagoburst). It was only the lowest KYNA dose that influenced the mitogenic response of lymphocytes, namely by increasing the proliferation of T cells. The impact on the phagocytic activity was varied with KYNA dose and administration time. However, all the KYNA doses significantly lowered the activity of oxidative burst in phagocytes, which was probably associated with its antioxidant properties. In the light of the research results, kynurenic acid may find applications as an immuno-modulating agent able to correct an excessive or insufficient response of phagocytizing cells, protecting an organism from oxidative stress.

  18. Dry olive leaf extract counteracts L-thyroxine-induced genotoxicity in human peripheral blood leukocytes in vitro.

    PubMed

    Topalovi?, Dijana Žukovec; Živkovi?, Lada; ?abarkapa, Andrea; Djeli?, Ninoslav; Baji?, Vladan; Dekanski, Dragana; Spremo-Potparevi?, Biljana

    2015-01-01

    The thyroid hormones change the rate of basal metabolism, modulating the consumption of oxygen and causing production of reactive oxygen species, which leads to the development of oxidative stress and DNA strand breaks. Olive (Olea europaea L.) leaf contains many potentially bioactive compounds, making it one of the most potent natural antioxidants. The objective of this study was to evaluate the genotoxicity of L-thyroxine and to investigate antioxidative and antigenotoxic potential of the standardized oleuropein-rich dry olive leaf extract (DOLE) against hydrogen peroxide and L-thyroxine-induced DNA damage in human peripheral blood leukocytes by using the comet assay. Various concentrations of the extract were tested with both DNA damage inducers, under two different experimental conditions, pretreatment and posttreatment. Results indicate that L-thyroxine exhibited genotoxic effect and that DOLE displayed protective effect against thyroxine-induced genotoxicity. The number of cells with DNA damage, was significantly reduced, in both pretreated and posttreated samples (P < 0.05). Comparing the beneficial effect of all tested concentrations of DOLE, in both experimental protocols, it appears that extract was more effective in reducing DNA damage in the pretreatment, exhibiting protective role against L-thyroxine effect. This feature of DOLE can be explained by its capacity to act as potent free radical scavenger. PMID:25789081

  19. Dry Olive Leaf Extract Counteracts L-Thyroxine-Induced Genotoxicity in Human Peripheral Blood Leukocytes In Vitro

    PubMed Central

    Žukovec Topalovi?, Dijana; Živkovi?, Lada; ?abarkapa, Andrea; Djeli?, Ninoslav; Baji?, Vladan; Spremo-Potparevi?, Biljana

    2015-01-01

    The thyroid hormones change the rate of basal metabolism, modulating the consumption of oxygen and causing production of reactive oxygen species, which leads to the development of oxidative stress and DNA strand breaks. Olive (Olea europaea L.) leaf contains many potentially bioactive compounds, making it one of the most potent natural antioxidants. The objective of this study was to evaluate the genotoxicity of L-thyroxine and to investigate antioxidative and antigenotoxic potential of the standardized oleuropein-rich dry olive leaf extract (DOLE) against hydrogen peroxide and L-thyroxine-induced DNA damage in human peripheral blood leukocytes by using the comet assay. Various concentrations of the extract were tested with both DNA damage inducers, under two different experimental conditions, pretreatment and posttreatment. Results indicate that L-thyroxine exhibited genotoxic effect and that DOLE displayed protective effect against thyroxine-induced genotoxicity. The number of cells with DNA damage, was significantly reduced, in both pretreated and posttreated samples (P < 0.05). Comparing the beneficial effect of all tested concentrations of DOLE, in both experimental protocols, it appears that extract was more effective in reducing DNA damage in the pretreatment, exhibiting protective role against L-thyroxine effect. This feature of DOLE can be explained by its capacity to act as potent free radical scavenger. PMID:25789081

  20. Human Recombinant Antibodies against Plasmodium falciparum Merozoite Surface Protein 3 Cloned from Peripheral Blood Leukocytes of Individuals with Immunity to Malaria Demonstrate Antiparasitic Properties

    Microsoft Academic Search

    Rasmus Lundquist; Leif Kofoed Nielsen; Ali Jafarshad; Daw SoeSoe; Lars Harder Christensen; P. Druilhe; M. H. Dziegiel

    2006-01-01

    Immunoglobulins from individuals with immunity to malaria have a strong antiparasitic effect when trans- ferred to Plasmodium falciparum malaria infected patients. One prominent target of antiparasitic antibodies is the merozoite surface antigen 3 (MSP-3). We have investigated the antibody response against MSP-3 residues 194 to 257 (MSP-3194-257) on the molecular level. mRNA from peripheral blood leukocytes from clinically immune individuals

  1. Psychological Mediation of a Type of Oxidative DNA Damage, 8-Hydroxydeoxyguanosine, in Peripheral Blood Leukocytes of Non-Smoking and Non-Drinking Workers

    Microsoft Academic Search

    Masahiro Irie; Shinya Asami; Shoji Nagata; Masakazu Miyata; Hiroshi Kasai

    2002-01-01

    Background: The present study investigates whether the formation of 8-hydroxydeoxyguanosine (8-OH-dG), a known oxidative DNA damage relevant to carcinogenicity, can be associated with psychological factors, in order to clarify the possible stress-cancer linkage from a genetic viewpoint. Methods: We performed a cross-sectional study in which we examined the relationships of the levels of 8-OH-dG in peripheral blood leukocytes to various

  2. In vitro study of interactions between silicon-containing nanoparticles and human peripheral blood leukocytes.

    PubMed

    Andreeva, E R; Rudimov, E G; Gornostaeva, A N; Beklemyshev, V I; Makhonin, I I; Maugeri, U O G; Buravkova, L B

    2013-07-01

    The effects of silicon dioxide-based nanoparticles on the viability and proliferative activity of human peripheral blood cultured lymphocytes were studied. All nanoparticles in a concentration of 100 ?g/ml produced a significant cytotoxic effect, its intensity depending on particles' structure: SiO2 nanoparticles were least toxic, while Ce3(+)-intercaled montmorillonite nanoparticles were most toxic. The cells died mainly by apoptosis and postapoptotic necrosis. Incubation with nanoparticles in a concentration of 100 ?g/ml for 72 h caused death of all phytohemagglutinin-activated lymphocytes, while in concentrations of 1 and 10 ?g/ml the nanoparticles had no effect of proliferative activity of cells. The results suggest that the effects of nanoparticles on cells are determined by the nanoparticle concentration and size, as well as by their structure. PMID:24137611

  3. The effect of injectable trace minerals (selenium, copper, zinc, and manganese) on peripheral blood leukocyte activity and serum superoxide dismutase activity of lactating Holstein cows.

    PubMed

    Machado, V S; Oikonomou, G; Lima, S F; Bicalho, M L S; Kacar, C; Foditsch, C; Felippe, M J; Gilbert, R O; Bicalho, R C

    2014-05-01

    The objective of this study was to evaluate the effect of subcutaneous supplementation of 300?mg of zinc, 50?mg of manganese, 25?mg of selenium, and 75?mg of copper on peripheral blood leukocyte activity and serum ?-hydroxybutyrate (BHBA) concentrations at 10?±?2 days in milk (DIM), and on serum superoxide dismutase (SOD) activity during the transition period and subsequent lactation of multiparous Holstein cows. A total of 250 multiparous cows were randomly allocated into one of two treatments groups, namely, trace mineral supplemented (TMS) or control. Cows in the TMS group were injected at 230 and 260 days of gestation, and 35 days postpartum. Serum SOD activity was measured at enrollment, and 10, 60 and 100 DIM. Serum BHBA concentration and leukocyte function were assessed at 10 DIM. Overall serum SOD activity for TMS and control was 16.01 and 12.71?U/mL, respectively. The interaction between treatment and time of serum collection was significant. Additionally, overall serum SOD activity was 12.85 and 14.78?U/mL for cows diagnosed with mastitis and unaffected cows, respectively. Treatment did not affect leukocyte function. For parity >2, TMS cows had lower serum BHBA concentrations than control cows; BHBA concentrations were 0.41 and 0.27?mmol/L for control and TMS cows, respectively. In conclusion, cows diagnosed with mastitis had decreased serum SOD activity, and trace mineral supplementation increased serum SOD activity although leukocyte function was not affected by supplementation. PMID:24685102

  4. Selective Activation of Cannabinoid Receptor 2 in Leukocytes Suppresses Their Engagement of the Brain Endothelium and Protects the Blood-Brain Barrier

    PubMed Central

    Rom, Slava; Zuluaga-Ramirez, Viviana; Dykstra, Holly; Reichenbach, Nancy L.; Pacher, Pal; Persidsky, Yuri

    2014-01-01

    Cannabinoid receptor 2 (CB2) is highly expressed in immune cells and stimulation decreases inflammatory responses. We tested the idea that selective CB2 activation in human monocytes suppresses their ability to engage the brain endothelium and migrate across the blood-brain barrier (BBB), preventing consequent injury. Intravital videomicroscopy was used to quantify adhesion of leukocytes to cortical vessels in lipopolysaccharide-induced neuroinflammation, after injection of ex vivo CB2–activated leukocytes into mice; CB2 agonists markedly decreased adhesion of ex vivo labeled cells in vivo. In an in vitro BBB model, CB2 activation in monocytes largely attenuated adhesion to and migration across monolayers of primary human brain microvascular endothelial cells and diminished BBB damage. CB2 stimulation in monocytes down-regulated active forms of integrins, lymphocyte function-associated antigen 1 (LFA-1), and very late antigen 4 (VLA-4). Cells treated with CB2 agonists exhibited increased phosphorylation levels of inhibitory sites of the actin-binding proteins cofilin and VASP, which are upstream regulators of conformational integrin changes. Up-regulated by relevant stimuli, Rac1 and RhoA were suppressed by CB2 agonists in monocytes. CB2 stimulation decreased formation of lamellipodia, which play a key role in monocyte migration. These results indicate that selective CB2 activation in leukocytes decreases key steps in monocyte–BBB engagement, thus suppressing inflammatory leukocyte responses and preventing neuroinflammation. PMID:24055259

  5. Synthesis and Secretion of Cystic Fibrosis Ciliary Dyskinesia Substances by Purified Subpopulations of Leukocytes

    PubMed Central

    Wilson, Gregory B.; Bahm, Valorie J.

    1980-01-01

    Cultured peripheral blood leukocytes (PBL) from individuals homozygous or heterozygous for the defective gene causing the inherited disease cystic fibrosis (CF) secrete three different ciliary dyskinesia substances (CDS), which can be detected by their activity in vitro in a rabbit mucociliary bioassay. Their PBL also release substances that promote mucus expulsion and destruction of the ciliated epithelium. In the present study the relative numbers of lymphocytes (T, B, and null), monocytes-macrophages (M?), and polymorphonuclear neutrophils were found to be normal in subjects with the CF gene, as were the responses of their PBL to phytohemagglutinin and pokeweed mitogen. Using purified subpopulations of leukocytes, we obtained evidence that both monocytes and T lymphocytes can secrete CDS in vitro with no requirement for cooperation with other lymphocyte subsets, whereas B and “null” lymphocytes probably require either differentiation or cellular cooperation for optimal secretion of CDS. Mucus expulsion and tissue destruction were produced by substances released primarily from polymorphonuclear neutrophils and secondarily from M?. Using cycloheximide and actinomycin D, we obtained evidence that CDS accumulation requires active protein synthesis and is not dependent on newly synthesized RNA, at least in short-term cultures. Gel filtration chromatography of active culture supernates showed that T lymphocytes synthesized only a CF-specific CDS, whereas M? synthesized all three CDS found in PBL cultures. Evidence is presented that one CDS is related structurally to C3a, since it can be removed with rabbit antisera specific for human C3a. PMID:7430342

  6. Calcium phosphate coating of nickel-titanium shape-memory alloys. Coating procedure and adherence of leukocytes and platelets.

    PubMed

    Choi, Jongsik; Bogdanski, Denise; Köller, Manfred; Esenwein, Stefan A; Müller, Dietmar; Muhr, Gert; Epple, Matthias

    2003-09-01

    Nickel-titanium shape-memory alloys (NiTi-SMA) were coated with calcium phosphate by dipping in oversaturated calcium phosphate solution. The layer thickness (typically 5-20 micrometer) can be varied by choice of the immersion time. The porous nature of the layer of microcrystals makes it mechanically stable enough to withstand both the shape-memory transition upon cooling and heating and also strong bending of the material (superelastic effect). This layer may improve the biocompatibility of NiTi-SMA, particulary for osteosynthetic devices by creating a more physiological surface and by restricting a potential nickel release. The adherence of human leukocytes (peripheral blood mononuclear cells and polymorphonuclear neutrophil granulocytes) and platelets to the calcium phosphate layer was analyzed in vitro. In comparison to non-coated NiTi-SMA, leukocytes and platelets showed a significantly increased adhesion to the coated NiTi-SMA. PMID:12818540

  7. Immunological characterization of peripheral blood leukocytes using vaccine for mycoplasmal pneumonia of swine (MPS) in swine line selected for resistance to MPS.

    PubMed

    Shimazu, Tomoyuki; Borjigin, Liushiqi; Katayama, Yuki; Li, Meihua; Satoh, Takumi; Watanabe, Kouichi; Kitazawa, Haruki; Roh, Sang-gun; Aso, Hisashi; Katoh, Kazuo; Suda, Yoshihito; Sakuma, Akiko; Nakajo, Mituru; Suzuki, Keiichi

    2013-10-01

    This study was conducted to evaluate immunological changes in peripheral blood leukocytes in pigs that were genetically selected for their improved resistance to mycoplasmal pneumonia of swine (MPS), using MPS vaccine as an antigen. Twelve castrated MPS-selected Landrace pigs were compared with the same number of pigs from a nonselected line by using a time-course analysis at the hematological level. After the second sensitization with MPS vaccine, the percentages of B cells, CD4(+) T cells, and natural killer (NK) cells in total leukocytes were lower in the selected line than in the nonselected line, whereas the percentage of granulocytes in total leukocytes increased in the MPS-selected line. We also assessed the proliferative ability of peripheral blood mononuclear cells (PBMCs) stimulated with Mycoplasma hyopneumoniae, lipopolysaccharide or concanavalin A, and found that although the proliferative ability of the PBMC was not different between the two lines at a steady state, the nonselected line showed a significantly higher proliferative ability after sensitization with MPS vaccine than the selected line regardless of antigens used. These results thus indicate that the selection of pigs on the basis of MPS resistance changes their immunophenotype, and would give us beneficial information for the prevention of MPS infection. PMID:23607374

  8. Biotin deficiency reduces expression of SLC19A3, a potential biotin transporter, in leukocytes from human blood.

    PubMed

    Vlasova, Tatyana I; Stratton, Shawna L; Wells, Amanda M; Mock, Nell I; Mock, Donald M

    2005-01-01

    In evaluating potential indicators of biotin status, we quantitated the expression of biotin-related genes in leukocytes from human blood of normal subjects before and after inducing marginal biotin deficiency. Biotin deficiency was induced experimentally by feeding an egg-white diet for 28 d. Gene expression was quantitated for the following biotin-related proteins: methylcrotonyl-CoA carboxylase chains A (MCCA) and B (MCCB); propionyl-CoA carboxylase chains A (PCCA) and B (PCCB); pyruvate carboxylase (PC); acetyl-CoA carboxylase isoforms A (ACCA) and B (ACCB); holocarboxylase synthetase (HCS); biotinidase; and 2 potential biotin transporters: sodium-dependent multivitamin transporter (SMVT) and solute carrier family 19 member 3 (SLC19A3). For 7 subjects who successfully completed the study, the abundance of the specific mRNAs was determined by quantitative real-time RT-PCR at d 0 and 28. At d 28, SLC19A3 expression had decreased to 33% of d 0 (P < 0.02 by two-tailed, paired t test). Expression of MCCA, PCCA, PC, ACCA, ACCB, HCS, biotinidase, and SMVT decreased to approximately 80% of d 0 (P < 0.05). Expression of the MCCB and PCCB chains that do not carry the biotin-binding motif did not change significantly; we speculate that expression of the biotin-binding chains of biotin-dependent carboxylases is more responsive to biotin status changes. These data provide evidence that expression of SLC19A3 is a relatively sensitive indicator of marginal biotin deficiency. PMID:15623830

  9. Activation of Cannabioid Receptor 2 Attenuates Leukocyte-Endothelial Interactions and Blood-Brain Barrier Dysfunction under Inflammatory Conditions

    PubMed Central

    Ramirez, Servio H.; Haskó, János; Skuba, Andrew; Fan, Shongshan; McCormick, Ryan; Dykstra, Holly; Reichenbach, Nancy; Krizbai, Istvan; Mahadevan, Anu; Zhang, Ming; Tuma, Ronald; Son, Young-jin; Persidsky, Yuri

    2012-01-01

    Previous studies have shown that modulation of the receptor-mediated cannabinoid system during neuroinflammation can produce potent neuroprotective and anti-inflammatory effects. However in this context, little is known about how selective activation of the cannabinoid type-2 receptor (CB2R) affects the activated state of the brain endothelium and blood brain barrier (BBB) function. Using human brain tissues and primary human brain endothelial cells (BMVEC) we demonstrate that the CB2R is highly upregulated during inflammatory insult. We then examined whether the CB2R agonists could attenuate inflammatory responses at the BBB using a mouse model of LPS-induced encephalitis and highly selective CB2R agonists. Visualization by intravital microscopy revealed that administration of JWH133 or a novel resorcinol-based compound O-1966, greatly attenuated leukocyte adhesion in surface pial vessels and in deep ascending cortical post-capillary venules. BBB permeability assessments with small and large fluorescent tracers showed that CB2R agonists were effective at preventing barrier leakiness after LPS administration. To determine whether the effects by CB2R agonists on barrier protection are not only due to the CB2R modulation of immune cell function, we tested the agonists in-vitro with barrier forming primary BMVEC. Remarkably, the addition of CB2R agonist increased trans-endothelial electrical resistance and increased the amount of tight junction protein present in membrane fractions. Furthermore, CB2R agonists decreased the induction of ICAM-1 and VCAM-1 surface expression in BMVEC exposed to various pro-inflammatory mediators. Together, these results suggest that pharmacological CB2R ligands offer a new strategy for BBB protection during neuroinflammation. PMID:22442067

  10. Integrin ?D?2 (CD11d/CD18) Is Expressed by Human Circulating and Tissue Myeloid Leukocytes and Mediates Inflammatory Signaling

    PubMed Central

    Miyazaki, Yasunari; Vieira-de-Abreu, Adriana; Harris, Estelle S.; Shah, Amrapali M.; Weyrich, Andrew S.; Castro-Faria-Neto, Hugo C.; Zimmerman, Guy A.

    2014-01-01

    Integrin ?D?2 is the most recently identified member of the leukocyte, or ?2, subfamily of integrin heterodimers. Its distribution and functions on human leukocytes have not been clearly defined and are controversial. We examined these issues and found that ?D?2 is prominently expressed by leukocytes in whole blood from healthy human subjects, including most polymorphonuclear leukocytes and monocytes. We also found that ?D?2 is displayed by leukocytes in the alveoli of uninjured and inflamed human lungs and by human monocyte-derived macrophages and dendritic cells, indicating broad myeloid expression. Using freshly-isolated human monocytes, we found that ?D?2 delivers outside-in signals to pathways that regulate cell spreading and gene expression. Screening expression analysis followed by validation of candidate transcripts demonstrated that engagement of ?D?2 induces mRNAs encoding inflammatory chemokines and cytokines and secretion of their protein products. Thus, ?D?2 is a major member of the integrin repertoire of both circulating and tissue myeloid leukocytes in humans. Its broad expression and capacity for outside-in signaling indicate that it is likely to have important functions in clinical syndromes of infection, inflammation, and tissue injury. PMID:25415295

  11. Chemiluminescence and superoxide anion production by leukocytes from chronic hemodialysis patients

    Microsoft Academic Search

    Elizabeth E Ritchey; John D Wallin; Sudhir V Shah

    1981-01-01

    Chemiluminescence and superoxide anion production by leukocytes from chronic hemodialysis patients. During phagocytosis or in response to a soluble stimulus, polymorphonuclear leukocytes (PMN) undergo a burst of oxidative metabolism involved intimately in antimicrobial activity. Superoxide anion produced during the burst is bactericidal either directly or as an intermediate metabolite. In addition, stimulated PMN's emit light or chemiluminescence (CL). CL is

  12. In Vitro Expression of Adhesion Receptors and Diapedesis by Polymorphonuclear Neutrophils during Experimentally Induced Streptococcus uberis Mastitis

    Microsoft Academic Search

    ELKE SMITS; CHRISTIAN BURVENICH; ALBERT J. GUIDRY; EDDY ROETS

    1998-01-01

    The expression of adhesion receptors and diapedesis by polymorphonuclear neutrophils (PMN) were studied before and during experimentally induced Streptococcus uberis mastitis. Both quarters of the left half of the ud- ders of five midlactation cows were inoculated with a suspension containing approximately 500 CFU of S. uberis 0140J. Clinical signs of an inflammatory reaction and leukocyte influx were observed 24

  13. Cytoplasmic strains and strain rates in motile polymorphonuclear leukocytes.

    PubMed Central

    Simon, S I; Schmid-Schönbein, G W

    1990-01-01

    A new method is presented to measure local cytoplasmic deformation and rate of deformation in motile active neutrophils. The deformation is expressed in terms of biomechanical strains and strain rates. For this purpose small phagocytosed latex microspheres were used as intracellular markers. Planar Lagrangian and Eulerian strains and the rate of strain were estimated from the positions of a triad of internalized markers. Principal strains, stretch ratios, and principal directions were computed. The intracellular strains were found to be large relative to the overall cell shape change. Principal cytoplasmic stretch ratios showed large extension in the direction of pseudopod formation and cell locomotion and contraction in perpendicular directions. Regional strain analysis showed contractile strains to predominate in the vicinity of the pseudopod or leading edge of motion. The transitional region between the pseudopod and the main cell body exhibited large shear strains. The posterior region, where the uropod is located, also revealed large extensions but small contractile strains. The rate of strains are relatively small, nonuniform in time, and largely independent of the strain. The method we propose to measure cytoplasmic strain can be applied to a variety of problems in cell mechanics. Images FIGURE 3 PMID:2207240

  14. Damage of human polymorphonuclear leukocytes by Junin virus

    Microsoft Academic Search

    R. P. Laguens; P. H. Gonzalez; C. Ponzinibbio; J. Chambo

    1986-01-01

    One of the most constant manifestations of Argentine Haemorrhagic Fever (AHF) is a marked leukopenia, which is also observed in experimental infection of guinea pigs [6] and monkeys [9] with pathogenic strains of Junin virus (JV), the etiologic agent of AHF. The pathogenic mechanism leading to leukopenia is at present poorly understood. Although destruction of lymphatic tissue, characteristic of lethal

  15. Expression of tlr4, md2 and cd14 in equine blood leukocytes during endotoxin infusion and in intestinal tissues from healthy horses.

    PubMed

    Fossum, C; Hjertner, B; Olofsson, K M; Lindberg, R; Ahooghalandari, P; Camargo, M M; Bröjer, J; Edner, A; Nostell, K

    2012-12-15

    The expression of tlr4, md2 and cd14 was studied in equine blood leukocytes and in intestinal samples using real time PCR. The stability of three commonly used reference genes, glyceraldehyde-3P-dehydrogenase (GAPDH), hypoxantine ribosyltransferase (HPRT) and succinate dehydrogenase complex subunit A (SDHA), was evaluated using qbase(PLUS). The equine peripheral blood mononuclear cells (eqPBMC) examined were either stimulated in vitro with Phorbol 12-myristate 13-acetate (PMA) and ionomycin or with the CpG oligodeoxynuclotide 2216 (CpG-ODN 2216) or obtained from horses before, during and after infusion of endotoxin. Intestinal tissue from healthy horses was sampled at ileum, right dorsal colon and rectum. Ranking of the three reference genes used for normalisation identified the combination HPRT/SDHA as most suitable both when determined ex vivo in leukocytes obtained from experimentally induced endotoxaemia and in eqPBMC activated in vitro while HPRT/GAPDH were most appropriate for the intestinal samples. The relative amounts of mRNA for TLR4 and MD-2 increased threefold during in vitro activation of the cells with CpG-ODN 2216 but was decreased in cultures stimulated with PMA/ionomycin. A transient elevation in the transcription of tlr4 and md2 was also evident for equine blood leukocytes following endotoxaemia. The levels of mRNA for CD14 on the other hand remained unaffected both during the induction of endotoxaemia and in the in vitro stimulated PBMCs. A low steady expression of TLR4, MD-2 and CD14 mRNA was demonstrated for the intestinal samples with no variation between the intestinal segments analysed. Thus, the foundation for real time PCR based levels of analysis of mRNA for all three components in the equine LPS receptor complex in different intestinal segments was set, making it possible to carry out future expression studies on clinical material. PMID:23036528

  16. Effect of peg-filgrastim-supported dose-dense adjuvant chemotherapy on the peripheral blood leukocyte phenotype in breast cancer patients.

    PubMed

    Collovŕ, Elena; Rovati, Bianca; Grasso, Donatella; Bencardino, Katia; Manzoni, Mariangela; Danova, Marco

    2009-01-01

    The aim of this study was to evaluate the effect of dose-dense adjuvant chemotherapy regimens with peg-filgrastim support on the phenotype of peripheral blood leukocytes in breast cancer patients. We evaluated the leukocyte phenotype of 14 patients aged 46-67 years undergoing 4 courses of chemotherapy with either epirubucin/cyclophosphamide (n=7) or 5-fluorouracil/epirubucin/cyclophosphamide (n=7) followed by 4 courses of taxol supported by peg-filgrastim (6 mg) administered 72 h after each chemotherapy course. The overall leukocyte number significantly increased from the first treatment course, while total lymphocytes tended to decrease with a negative peak following the 6th course (p=0.03). B (CD19+, CD20+) and early B lymphocyte subsets (CD20+/CD38+) significantly decreased during treatment (p<0.05), while T lymphocyte subsets did not show significant changes, except a decrease in T helper (CD4+) cells. Immature T lymphocytes (CD4+/CD8+ subset), dendritic cells (CD11c+) and NK cells (CD56+) increased with respect to the baseline. Our results suggest that dose-dense chemotherapy programs with the support of peg-filgrastim did not significantly impair the immune system of breast cancer patients and allowed for a rapid restoration of most immune competent cells. These observations may have important clinical implications with a view to vaccination or other immunotherapeutic approaches to solid tumours. PMID:21475795

  17. Neuroimmunity and the Blood–Brain Barrier: Molecular Regulation of Leukocyte Transmigration and Viral Entry into the Nervous System with a Focus on NeuroAIDS

    PubMed Central

    Buckner, Clarisa M.; Luers, Aimée J.; Calderon, Tina M.; Eugenin, Eliseo A.

    2015-01-01

    HIV infection of the central nervous system (CNS) can result in neurologic dysfunction with devastating consequences in a significant number of individuals with AIDS. Two main CNS complications in individuals with HIV are encephalitis and dementia, which are characterized by leukocyte infiltration into the CNS, microglia activation, aberrant chemokine expression, blood–brain barrier (BBB) disruption, and eventual damage and/or loss of neurons. One of the major mediators of NeuroAIDS is the transmigration of HIV-infected leukocytes across the BBB into the CNS. This review summarizes new key findings that support a critical role of the BBB in regulating leukocyte transmigration. In addition, we discuss studies on communication among cells of the immune system, BBB, and the CNS parenchyma, and suggest how these interactions contribute to the pathogenesis of Neuro-AIDS. We also describe some of the animal models that have been used to study and characterize important mechanisms that have been proposed to be involved in HIV-induced CNS dysfunction. Finally, we review the pharmacologic interventions that address neuroinflammation, and the effect of substance abuse on HIV-1 related neuroimmunity. PMID:18040782

  18. Mononuclear leukocyte infiltrate in extraplacental membranes and preterm delivery.

    PubMed

    Holzman, Claudia; Senagore, Patricia K; Wang, Jianling

    2013-05-15

    Large numbers of polymorphonuclear leukocytes in the amnion and chorion define histological chorioamnionitis (HCA), a condition linked to spontaneous preterm delivery (PTD). Less is known about placental patterns of mononuclear leukocyte (MNL) density and PTD. In this prospective study (1998-2004), women were sampled from 52 clinics in 5 Michigan communities and enrolled at 16-27 weeks' gestation. HCA and MNL distributions in delivered placentas were evaluated microscopically in a subcohort (290 preterm, 823 term). Midpregnancy biomarkers from maternal blood (i.e., C-reactive protein (CRP), corticotropin-releasing hormone, and cytokines) were compared among term and PTD subjects grouped by presence/absence of HCA and high MNL density. A density of more than 10 MNLs per high-power field in the chorion of the membrane roll, referred to as MNL-CMR, was associated with medically indicated PTD (odds ratio = 2.2, 95% confidence interval: 1.3, 3.6) and spontaneous PTD (odds ratio = 2.5, 95% confidence interval: 1.7, 3.7). Associations persisted after removal of women with HCA-positive placentas, abruption, hypertensive disorders, or obesity. HCA-associated PTD showed higher CRP and cytokine levels. MNL-CMR-associated PTD showed higher CRP and corticotropin-releasing hormone levels. These data suggest that an MNL infiltrate in the chorion of the membrane roll marks PTD pathways that are distinct from HCA and not entirely explained by pregnancy complications. PMID:23429723

  19. Prothrombotic roles of substance-P, neurokinin-1 receptors and leukocytes in the platelet-dependent clot formation in whole blood

    Microsoft Academic Search

    Toshiharu Azma; Yuki Matsubara; Hiroyuki Kinoshita; Ikuhiro Hidaka; Seiji Shiraishi; Masakazu Nakao; Masashi Kawamoto; Osafumi Yuge; Yoshio Hatano

    2009-01-01

    A number of types of non-neuronal cells including leukocytes have been confirmed to possess substance-P and its specific neurokinin-1\\u000a receptor (NK1R), while the pathophysiological roles of substance-P in these cells remain to be established. Effects of substance-P\\u000a through NK1R on platelet-dependent clot formation were evaluated by using an oscillating-probe viscoelastometer. The clot\\u000a signal, indicative of the clot strength in blood-derived

  20. Effects of Cigarette Smoke Fractions on the Chemotaxis of Polymorphonuclear Leu kocytes

    Microsoft Academic Search

    Raymond B. Bridges; Lily Hsieh

    The effects of cigarette smoking fractions on polymorphonuclear leukocyte (PMN) chemotaxis were determined using the 51Cr-assay. Water-soluble frac- tions (WSF) of cigarette smoke produced from several tobacco types differed in inhibitory potencies (i.e., flue curedMarylandblended > Burley ? Turkish) corresponding to the respective unsaturated aldehyde content of the smoke from these tobaccos. Fractionation of cigarette smoke condensate (CSC) demonstrated that

  1. Hesperidin Displays Relevant Role in the Nutrigenomic Effect of Orange Juice on Blood Leukocytes in Human Volunteers: A Randomized Controlled Cross-Over Study

    PubMed Central

    Milenkovic, Dragan; Deval, Christiane; Dubray, Claude; Mazur, Andrzej; Morand, Christine

    2011-01-01

    Background We previously showed, in healthy, middle-aged, moderately overweight men, that orange juice decreases diastolic blood pressure and significantly improves postprandial microvascular endothelial reactivity and that hesperidin could be causally linked to the observed beneficial effect of orange juice. The objective was to determine the effect of chronic consumption of orange juice on the gene expression profile of leukocytes in healthy volunteers and to assess to what extent hesperidin is involved in the effect of orange juice. Methodology/Principal Findings Volunteers were included in a randomized, controlled, crossover study. Throughout three 4-week periods, volunteers consumed daily: 500 ml orange juice, 500 ml control drink plus hesperidin or 500 ml control drink and placebo. Blood samplings were performed on 10 overnight-fasted subjects after the 4-week treatment period. Global gene expression profiles were determined using human whole genome cDNA microarrays. Both orange juice and hesperidin consumption significantly affected leukocyte gene expression. Orange juice consumption induced changes in expression of, 3,422 genes, while hesperidin intake modulated the expression of 1,819 genes. Between the orange juice and hesperidin consumption groups, 1,582 regulated genes were in common. Many of these genes are implicated in chemotaxis, adhesion, infiltration and lipid transport, which is suggestive of lower recruitment and infiltration of circulating cells to vascular wall and lower lipid accumulation. Conclusions This study shows that regular consumption of orange juice for 4 weeks alters leukocyte gene expression to an anti-inflammatory and anti-atherogenic profile, and hesperidin displays a relevant role in the genomic effect of this beverage. Trial Registration ClinicalTrials.gov NCT 00983086 PMID:22110589

  2. Label-free in vivo imaging of human leukocytes using two-photon excited endogenous fluorescence

    NASA Astrophysics Data System (ADS)

    Zeng, Yan; Yan, Bo; Sun, Qiqi; Teh, Seng Khoon; Zhang, Wei; Wen, Zilong; Qu, Jianan Y.

    2013-04-01

    We demonstrate that two-photon excited endogenous fluorescence enables label-free morphological and functional imaging of various human blood cells. Specifically, we achieved distinctive morphological contrast to visualize morphology of important leukocytes, such as polymorphonuclear structure of granulocyte and mononuclear feature of agranulocyte, through the employment of the reduced nicotinamide adenine dinucleotide (NADH) fluorescence signals. In addition, NADH fluorescence images clearly reveal the morphological transformation process of neutrophils during disease-causing bacterial infection. Our findings also show that time-resolved NADH fluorescence can be potentially used for functional imaging of the phagocytosis of pathogens by leukocytes (neutrophils) in vivo. In particular, we found that free-to-bound NADH ratios measured in infected neutrophils increased significantly, which is consistent with a previous study that the energy consumed in the phagocytosis of neutrophils is mainly generated through the glycolysis pathway that leads to the accumulation of free NADH. Future work will focus on further developing and applying label-free imaging technology to investigate leukocyte-related diseases and disorders.

  3. Enhanced Chemotactic and Phagocytic Activities of Leukocytes in Psoriasis Vulgaris

    Microsoft Academic Search

    A. Wahba; H. A. Cohen; M. Bar-Eli; R. Gallily

    1978-01-01

    Leukocytes derived from the peripheral blood of peripheral patients demonstrated an enhanced chemotactic response compared with leukocytes from healthy subjects. No significant difference was detected between the chemotactic response of leukocytes from patients with minimal or no skin involvement and those from patients with extensive lesions. Psoriatic leukocytes also had a significantly higher capacity to engulf 125I labeled Shigella flexneri

  4. Forces on a Wall-Bound Leukocyte in a Small Vessel Due to Red Cells in the Blood Stream

    E-print Network

    Freund, Jonathan B.

    exerted by the flowing blood. This force is particularly difficult to estimate due to the particulate. Effects of the particulate character of blood have been studied in detail for flow in small vessels (1­11), rolling of leuko- cytes (6,12), dynamics of vascular networks (13), and the design of blood microfluidic

  5. Effect of infection with BHV-1 on peripheral blood leukocytes and lymphocyte subpopulations in calves with subclinical BVD.

    PubMed

    Molina, V; Risalde, M A; Sánchez-Cordón, P J; Pedrera, M; Romero-Palomo, F; Luzzago, C; Gómez-Villamandos, J C

    2013-08-01

    Bovine viral diarrhea virus (BVDV) and bovine herpesvirus-1 (BHV-1) are important cattle pathogens that induce a broad immunosuppression on cell-mediated immune response on its own participating in the bovine respiratory disease complex (BRDC). The aim of our study was to evaluate the quantitative changes in immunocompetent cells in healthy calves and calves with subclinical bovine viral diarrhea (BVD), both inoculated with BHV-1. Total leukocyte counts exhibited changes mainly in neutrophils and lymphocytes that can contribute to the BVDV immunosuppression, thus accounting for some of the intergroup differences. Monocytes did not display numerical changes in either group. Regarding lymphocyte subpopulations, even though CD4+ T lymphocytes and B cells were depleted around 4 dpi in both infected groups, the main difference observed between both groups was in CD8+ T cells which displayed an earlier depletion in BVDV inoculated calves that can promote a greater BHV-1 dissemination, thus aggravating the course of the disease. PMID:23541923

  6. Impaired polymorphonuclear leucocyte chemotaxis in rheumatoid arthritis.

    PubMed Central

    Goddard, D H; Kirk, A P; Kirwan, J R; Johnson, G D; Holborow, E J

    1984-01-01

    This study has investigated the chemotactic activity of polymorphonuclear cells (PMNs) isolated from the blood of patients with either articular rheumatoid arthritis (RA) or RA with extra-articular manifestations. A double fluorochrome immunofluorescent staining test has been employed to identify cell-associated immunoglobulins, probably immune complexes. The results suggest an inverse relationship between PMN chemotaxis and staining for cell-associated immunoglobulins, either surface bound or internalised. PMNs from RA patients showed reduced chemotaxis, and this was further reduced when RA PMNs were incubated for 30 minutes in autologous serum. A similar reduction in chemotaxis of normal PMNs occurred after incubation in RA sera. Preincubation of both RA and normal PMNs in RA serum (but not normal serum) resulted in an increase in the number of cells in which cell-associated immunoglobulins were demonstrable. This further reduction in RA PMN chemotaxis after exposure to autologous serum, together with an increase in immunoglobulin staining, may indicate selection of certain PMNs at the time of venepuncture due to cell margination. Such a selection process would call for a re-evaluation of previous studies of RA PMN function in relation to the disease process. PMID:6324704

  7. Modification of radiation-induced DNA double strand break repair pathways by chemicals extracted from Podophyllum hexandrum: an in vitro study in human blood leukocytes.

    PubMed

    Srivastava, Nitya N; Shukla, Sandeep K; Yashavarddhan, M H; Devi, Memita; Tripathi, Rajendra P; Gupta, Manju L

    2014-06-01

    Radiation exposure is a serious threat to biomolecules, particularly DNA, proteins and lipids. Various exogenous substances have been reported to protect these biomolecules. In this study we explored the effect of pre-treatment with G-002M, a mixture of three active derivatives isolated from the rhizomes of Podophyllum hexandrum, on DNA damage response in irradiated human blood leukocytes. Blood was collected from healthy male volunteers, preincubated with G-002M and then irradiated with various doses of radiation. Samples were analyzed using flow cytometry to quantify DNA double strand break (DSB) biomarkers including ?-H2AX, P53BP1 and levels of ligase IV. Blood samples were irradiated in vitro and processed to determine time and dose-dependent kinetics. Semiquantitative RT-PCR was performed at various time points to measure gene expression of DNA-PKcs, Ku80, ATM, and 53BP1; each of these genes is involved in DNA repair signaling. Pre-treatment of blood with G-002M resulted in reduction of ?-H2AX and P53BP1 biomarkers levels and elevated ligase IV levels relative to non-G-002M-treated irradiated cells. These results confirm suppression in radiation-induced DNA DSBs. Samples pre-treated with G-002M and then irradiated also showed significant up-regulation of DNA-PKcs and Ku80 and downregulation of ATM and 53BP1 gene expressions, suggesting that G-002M plays a protective role against DNA damage. The protective effect of G-002M may be due to its ability to scavange radiation-induced free radicals or assist in DNA repair. Further studies are needed to decipher the role of G-002M on signaling molecules involved in radiation-induced DNA damage repair pathways. PMID:24500925

  8. Cytokine profiles of cord and adult blood leukocytes: differences in expression are due to differences in expression and activation of transcription factors

    PubMed Central

    Nitsche, Andreas; Zhang, Meixia; Clauss, Theresa; Siegert, Wolfgang; Brune, Kay; Pahl, Andreas

    2007-01-01

    Background Stem cell transplantation as therapy for hematological disorders is often hampered by severe graft-versus-host-disease. This may be reduced by umbilical cord blood transplantation, an effect that has been attributed to qualitative differences between neonatal and adult T cells. We compared levels of secreted proteins and cytokine mRNA induced in cord blood leukocytes (CBL) and adult blood leukocytes (ABL) by various stimuli. Results While interleukin-2 (IL-2) levels were similar in CBL and ABL, there was less induction of the Th1 cytokine interferon-? in CBL. Production of the Th2 cytokines IL-4, IL-5, and IL-13 and the hematopoietic cytokine IL-3 was much lower in CBL versus ABL after T-cell receptor-mediated stimulation, whereas production of GM-CSF was comparable in the 2 cell types. The lower levels of Th1 and Th2 cytokines were maintained in CBL during a 4-day time-course study, while after 12 hours IL-3 and GM-CSF reached in CBL levels similar to those in ABL. For all cytokines except IFN?, the IC50 values for inhibition by cyclosporin A were similar in ABL and CBL. In contrast, there was less expression and activation of transcription factors in CBL. Activation of NF-?B by TPA/ionomycin was detected in ABL but not CBL. Furthermore, there was less expression of the Th subset-specific transcription factors T-bet and c-maf in CBL versus ABL, whereas GATA-3 expression was similar. Expression of T-bet and c-maf correlated with expression of the Th1 and Th2 cytokines, respectively. Time course experiments revealed that T-bet expression was stimulated in both cell types, whereas c-maf and GATA-3 were induced only in ABL. Conclusion The diminished capability of CBL to synthesize cytokines is probably due to decreased activation of NF-?B, whereas differences in Th subsets are due to differences in regulation of Th lineage-specific transcriptions factors. We propose that the reduced incidence and severity of GvHD after allogeneic transplantation of umbilical CB cells is due to lesser activation of specific transcription factors and a subsequent reduction in production of certain cytokines. PMID:17764543

  9. Functions and oxidative stress status of leukocytes in patients with nephrotic syndrome

    Microsoft Academic Search

    Taner Akyol; Fatih Bulucu; Osman Sener; Levent Yamanel; Ahmet Aydin; Volkan Inal; Ergun Bozoglu; Erkan Demirkaya; Ayse Eken; Ugur Musabak

    2007-01-01

    This study was conducted to establish the functions and oxidative stress status in leukocytes of adult patients with nephrotic\\u000a syndrome. Thirty adult patients with nephrotic syndrome and 32 controls were included. Phagocytosis ability, the killing ability\\u000a of the micro-organism phagosited of polymorphonuclear leukocytes (PMNL) and monocytes, along with oxidative stress parameters\\u000a of PMNLs were assessed. There was no statistically significant

  10. PCR-based human leukocyte antigen (HLA) DQ alpha typing of blood stained light and dark blue denim fabric.

    PubMed

    Del Rio, S A; Marino, M A; Belgrader, P

    1996-05-01

    Obtaining typable PCR products from DNA purified from blood stained blue denim has been difficult since inhibitors of PCR in blue denim apparently co-purify with the DNA. Organic and chelex extraction methods were tested for their ability to purify typable DNA from either light or dark blue denim fabric both stained with blood. DNA purified from the light blue denim using either method was successfully used in obtaining correct HLA-DQ alpha typing results. The chelex, but not the organic, procedure was able to yield typable DNA when the dark blue denim was the substrate. Therefore, the chelex method may be more effective than the organic method in preventing compounds that inhibit PCR from co-purifying with the DNA. PMID:8656191

  11. Leukocyte transendothelial migration: A junctional affair

    Microsoft Academic Search

    Francis W Luscinskas; Shuo Ma; Asma Nusrat; Charles A Parkos; Sunil K Shaw

    2002-01-01

    A critical function of the inflammatory response is delivery of leukocytes to a site of injury, immune reaction or infection. Considerable information is available concerning the molecular mechanisms that capture flowing leukocytes and initiate their stable arrest on the lumenal surface of the blood vessel wall. In comparison, much less is known about the subsequent step(s) in migration of circulating

  12. Leukocyte recruitment to airways by aldehyde-carbon combinations that mimic cigarette smoke

    Microsoft Academic Search

    K. H. Kilburn; W. N. McKenzie

    1978-01-01

    Exposure of Syrian golden hamsters to formaldehyde (3 to 250 ppM) evaporated onto carbon (21 to 805 mg per cu. m) recruited polymorphonuclear (PMN) leukocytes to the epithelium of tracheas and intrapulmonary airways which peaked at 24 to 48 hours. Acrolein (< 6 ppM) on carbon 593 mg per cu. m) caused PMN recruitment which was maximal at 12 hours.

  13. Identification of an Additional Class of C3Binding Membrane Proteins of Human Peripheral Blood Leukocytes and Cell Lines

    Microsoft Academic Search

    Joe L. Cole; George A. Housley; Thomas R. Dykman; Richard P. MacDermott; John P. Atkinson

    1985-01-01

    Proteins binding the third component of complement (C3) were isolated by affinity chromatography from surface-labeled solubilized membranes of human peripheral blood cells and cell lines. The isolated molecules were subjected to NaDodSO4\\/PAGE, and autoradiographs of these gels indicated that C3-binding proteins could be divided into three groups based on Mr: (i) gp200, an ≈ 200,000 Mr molecule previously identified as

  14. Survival after transplantation of unrelated donor umbilical cord blood is comparable to that of human leukocyte antigen-matched unrelated donor bone marrow: results of a matched-pair analysis

    Microsoft Academic Search

    Juliet N. Barker; Stella M. Davies; Todd DeFor; Norma K. C. Ramsay; Daniel J. Weisdorf; John E. Wagner

    2001-01-01

    Umbilical cord blood (UCB) is being in- creasingly used for hematopoietic stem cell transplantation and has been associ- ated with a reduced incidence of severe graft-versus-host disease (GVHD). To fur- ther investigate the relative merits of unre- lated donor UCB versus bone marrow (BM), a matched-pair analysis comparing the outcomes of recipients of 0 to 3 human leukocyte antigen (HLA)-mis-

  15. Determination of acid alpha-naphthyl acetate esterase enzyme activity in peripheral blood leukocytes of gazelles (Gazella subgutturosa).

    PubMed

    Altunay, H; Harem, I S; Harem, M K; Asti, R N; Kurtdede, N

    2008-12-01

    We examined gazelle peripheral blood leucocytes using the alpha-Naphthyl acetate esterase (ANAE) staining technique (pH 5.8). Our purpose was to determine the percentage of ANAE positive lymphocytes. The proportion of ANAE positive T-lymphocytes was 72%. T-lymphocytes showed an ANAE positive reaction, but eosinophilic granulocytes and monocytes also showed a positive reaction. By contrast, no reaction was detected in B-lymphocytes, neutrophil granulocytes or platelets. The reaction observed in T-lymphocytes was a red-brown coloration, usually 1-2 granules, but enough granules to fill the cytoplasm were detected rarely. As a result of ANAE enzyme staining, we concluded that the staining technique can be used as a cytochemical marker for gazelle T-lymphocytes. PMID:19085516

  16. A melanoma helper peptide vaccine increases Th1 cytokine production by leukocytes in peripheral blood and immunized lymph nodes

    PubMed Central

    2014-01-01

    Background Cancers produce soluble and cell-associated molecules that can suppress or alter antitumor immunity. Preclinical studies suggest the disease burden may alter the cytokine profile of helper T cell responses to cancer antigens. We studied cytokine production by helper T cells responding to vaccination with 6 melanoma helper peptides (6MHP) in blood and lymph nodes. Methods Twenty-three patients with stage IIIB-IV melanoma received a 6MHP vaccine. Antigen-reactive T cells from blood and draining lymph nodes were cultured, exposed to antigen, and then supernatants (days 2 and 5) were assayed for Th1 and Th2 cytokines. Results from 4 time points were compared to pre-vaccine levels. Results Cytokine responses to vaccinating peptides were observed in 83% of patients. Th1 favoring responses were most common (17 of 19 responders). The most abundant cytokines produced were IFN-? and IL-5 in the PBMC’s. IL-2 responses predominated in cells obtained from draining lymph nodes in 2-day culture but not in 5-day cultures. Patients with clinically measurable disease produced similar levels of total cytokine and similar degree of Th1 polarization as patients with no evidence of disease (NED). Conclusions The MHC class II-associated peptides used in this study induced helper T cells with a Th1-biased cytokine response in both PBMC and sentinel immunized nodes. Most patients can mount a Th1 dominant response to these peptides. Future studies are needed to test newer vaccine adjuvants in combination with these peptides. Trial registration CDR0000378171, Clinicaltrials: NCT00089219. PMID:25126421

  17. Purinergic signaling gene network expression in bovine polymorphonuclear neutrophils during the peripartal period.

    PubMed

    Seo, J; Osorio, J S; Loor, J J

    2013-12-01

    An effective immune response relies on efficient activation of polymorphonuclear neutrophilic leukocytes (PMNL). The PMNL release cellular ATP in response to inflammatory mediators. Although extracellular ATP is rapidly degraded to adenosine, both compounds can readily bind to either the purinergic receptor P1 (adenosine) or P2 (ATP). The P1 and P2 receptors are members of the G-protein-coupled receptor family. The peripartal period is characterized by marked changes in metabolic and inflammatory status that are functionally related with immune responses in the cow. We evaluated the mRNA expression of genes associated with purinergic signaling in PMNL during the peripartal period. Seven multiparous Holstein cows were dried off at d -50 relative to expected parturition and fed a controlled-energy diet (net energy for lactation=1.24 Mcal/kg of dry matter) for ad libitum intake during the entire dry period. After calving, all cows were fed a common lactation diet (net energy for lactation=1.65 Mcal/kg of dry matter) until 30 d in milk. Blood PMNL collected at -10, 3, and 21 d in milk were used to study the expression of 22 genes associated with adhesion to endothelium, chemoattractant binding at the plasma membrane, and purinergic signaling. Other blood samples around calving were used to analyze concentrations of insulin, metabolites, and whole-blood phagocytosis. The expression of purinergic receptor P2Y, G-protein coupled, 2 (P2RY2) increased on d 3 and then decreased on d 21. This response suggested that ATP could play a role in the amplification of chemotactic signals. In contrast, the expression of genes encoding cell adhesion [selectin L (SELL) and selectin P ligand (SELPLG)], chemoattractant receptors [complement component 5a receptor 1 (C5AR1), IL-8 receptor ? (CXCR1), IL-8 receptor ? (CXCR2), and platelet-activating factor receptor (PTAFR)], and adenosine receptors [adenosine A1 receptor (ADORA1) and adenosine A3 receptor (ADORA3)] decreased between -10 and 3 d. The decrease coincided with a marked increase in blood nonesterified fatty acids and hydroxybutyrate concentrations, and a decrease in glucose and insulin concentrations. The increase in metabolites also was associated with greater expression of leukotriene B4 receptor (LTB4R) on d 3 and 21 compared with d -10, which is involved in inflammatory prostaglandin synthesis. Most chemoattractant receptors increased by 21 d, but cell adhesion genes and blood leukocyte phagocytosis was lower. The expression of adenosine A2a receptor (ADORA2A), which is associated with immunosuppression of PMNL and that of adenosine uptake channels [solute carrier family 29 (nucleoside transporters), member 1 (SLC29A1) and member 2 (SLC29A2)] and the nucleotidase adenosine deaminase (ADA) was greater at 3 and 21 d compared with -10d. The reduction in key immune responses, such as cell adhesion and chemotaxis, by bovine PMNL could partly be a function of changes in mRNA expression of genes associated with purinergic signaling. PMID:24119811

  18. Disorders of sex development expose transcriptional autonomy of genetic sex and androgen-programmed hormonal sex in human blood leukocytes

    PubMed Central

    Holterhus, Paul-Martin; Bebermeier, Jan-Hendrik; Werner, Ralf; Demeter, Janos; Richter-Unruh, Annette; Cario, Gunnar; Appari, Mahesh; Siebert, Reiner; Riepe, Felix; Brooks, James D; Hiort, Olaf

    2009-01-01

    Background Gender appears to be determined by independent programs controlled by the sex-chromosomes and by androgen-dependent programming during embryonic development. To enable experimental dissection of these components in the human, we performed genome-wide profiling of the transcriptomes of peripheral blood mononuclear cells (PBMC) in patients with rare defined "disorders of sex development" (DSD, e.g., 46, XY-females due to defective androgen biosynthesis) compared to normal 46, XY-males and 46, XX-females. Results A discrete set of transcripts was directly correlated with XY or XX genotypes in all individuals independent of male or female phenotype of the external genitalia. However, a significantly larger gene set in the PBMC only reflected the degree of external genital masculinization independent of the sex chromosomes and independent of concurrent post-natal sex steroid hormone levels. Consequently, the architecture of the transcriptional PBMC-"sexes" was either male, female or even "intersex" with a discordant alignment of the DSD individuals' genetic and hormonal sex signatures. Conclusion A significant fraction of gene expression differences between males and females in the human appears to have its roots in early embryogenesis and is not only caused by sex chromosomes but also by long-term sex-specific hormonal programming due to presence or absence of androgen during the time of external genital masculinization. Genetic sex and the androgen milieu during embryonic development might therefore independently modulate functional traits, phenotype and diseases associated with male or female gender as well as with DSD conditions. PMID:19570224

  19. Separation of uncompromised whole blood mixtures for single source STR profiling using fluorescently-labeled human leukocyte antigen (HLA) probes and fluorescence activated cell sorting (FACS).

    PubMed

    Dean, Lee; Kwon, Ye Jin; Philpott, M Katherine; Stanciu, Cristina E; Seashols-Williams, Sarah J; Dawson Cruz, Tracey; Sturgill, Jamie; Ehrhardt, Christopher J

    2015-07-01

    Analysis of biological mixtures is a significant problem for forensic laboratories, particularly when the mixture contains only one cell type. Contributions from multiple individuals to biologic evidence can complicate DNA profile interpretation and often lead to a reduction in the probative value of DNA evidence or worse, its total loss. To address this, we have utilized an analytical technique that exploits the intrinsic immunological variation among individuals to physically separate cells from different sources in a mixture prior to DNA profiling. Specifically, we applied a fluorescently labeled antibody probe to selectively bind to one contributor in a mixture through allele-specific interactions with human leukocyte antigen (HLA) proteins that are expressed on the surfaces of most nucleated cells. Once the contributor's cells were bound to the probe, they were isolated from the mixture using fluorescence activated cell sorting (FACS)-a high throughput technique for separating cell populations based on their optical properties-and then subjected to STR analysis. We tested this approach on two-person and four-person whole blood mixtures where one contributor possessed an HLA allele (A*02) that was not shared by other contributors to the mixture. Results showed that hybridization of the mixture with a fluorescently-labeled antibody probe complimentary to the A*02 allele's protein product created a cell population with a distinct optical profile that could be easily differentiated from other cells in the mixture. After sorting the cells with FACS, genetic analysis showed that the STR profile of this cell population was consistent with that of the contributor who possessed the A*02 allele. Minor peaks from the A*02 negative contributor(s) were observed but could be easily distinguished from the profile generated from A*02 positive cells. Overall, this indicates that HLA antibody probes coupled to FACS may be an effective approach for generating STR profiles of individual contributors from forensic mixtures. PMID:25796046

  20. The role of G-CSF and IL-6 in the granulopoiesis-stimulating activity of murine blood serum induced by perorally administered ultrafiltered pig leukocyte extract, IMUNOR.

    PubMed

    Vacek, Antonín; Hofer, Michal; Holá, Jirina; Weiterová, Lenka; Streitová, Denisa; Svoboda, Jaroslav

    2007-05-01

    IMUNOR, a low-molecular weight (< 12 kD) ultrafiltered pig leukocyte extract, has been previously found to have significant stimulatory effects on murine hematopoiesis supressed by ionizing radiation or cytotoxic drugs. This communication shows data on the mechanisms of these effects. Using ELISA assay, significantly increased levels of granulocyte colony-stimulating factor (G-CSF) and interleukin-6 (IL-6) were observed. On the contrary, no detectable levels of granulocyte-macrophage colony-stimulating factor (GM-CFC) and interleukin-3 (IL-3) have been found in blood serum of IMUNOR-treated mice. Incubation of the serum from IMUNOR-treated mice with antibodies against G-CSF caused abrogation of the ability of the sera to stimulate in vitro growth of colonies originating from granulocyte-macrophage progenitor cells (GM-CFC). In contrast, incubation of the serum with antibodies against IL-6 did not change its colony-stimulating activity. It may be inferred from these findings that G-CSF is probably the main cytokine responsible for the granulopoiesis-stimulating effects of IMUNOR. When the serum from IMUNOR-treated mice with G-CSF inactivated by anti-G-CSF antibodies (but with elevated IL-6) was added to cultures of bone marrow cells together with a suboptimum concentration of IL-3, a significant increase in the numbers of GM-CFC colonies was found. Moreover, conjoint inactivation of G-CSF and IL-6 significantly decreased the numbers of GM-CFC colonies in comparison with those observed when only G-CSF was inactivated. This observation strongly suggests that though IMUNOR-induced IL-6 is not able to induce the growth of GM-CFC colonies alone, it is able to potentiate the hematopoiesis-stimulating effect of IL-3. These findings represent a new knowledge concerning the hematopoiesis-stimulating action of IMUNOR, a promising immunomodulatory agent. PMID:17386413

  1. ?M?2-integrin-intercellular adhesion molecule-1 interactions drive the flow-dependent trafficking of Guillain-Barré syndrome patient derived mononuclear leukocytes at the blood-nerve barrier in vitro

    PubMed Central

    Yosef, Nejla; Ubogu, Eroboghene E.

    2012-01-01

    The mechanisms of hematogenous leukocyte trafficking at the human blood-nerve barrier (BNB) are largely unknown. Intercellular adhesion molecule-1 (ICAM-1) has been implicated in the pathogenesis of Guillain-Barré syndrome (GBS). We developed a cytokine-activated human in vitro BNB model using primary endoneurial endothelial cells. Endothelial treatment with 10 U/mL tissue necrosis factor-? and 20 U/mL interferon-? resulted in de novo expression of proinflammatory chemokines CCL2, CXCL9, CXCL11 and CCL20, with increased expression of CXCL2-3, CXCL8 and CXCL10 relative to basal levels. Cytokine treatment induced/ enhanced ICAM-1, E- and P-selectin, vascular cell adhesion molecule-1 and the alternatively spliced pro-adhesive fibronectin variant, fibronectin connecting segment-1 expression in a time-dependent manner, without alterations in junctional adhesion molecule-A expression. Lymphocytes and monocytes from untreated GBS patients express ICAM-1 counterligands, ?M- and ?L-integrin, with differential regulation of ?M-integrin expression compared to healthy controls. Under flow conditions that mimic capillary hemodynamics in vivo, there was a >3-fold increase in total GBS patient and healthy control mononuclear leukocyte adhesion/ migration at the BNB following cytokine treatment relative to the untreated state. Function neutralizing monoclonal antibodies against human ?M-integrin (CD11b) and ICAM-1 reduced untreated GBS patient mononuclear leukocyte trafficking at the BNB by 59% and 64.2% respectively. Monoclonal antibodies against ?L-integrin (CD11a) and human intravenous immunoglobulin reduced total leukocyte adhesion/migration by 22.8% and 17.6% respectively. This study demonstrates differential regulation of ?M-integrin on circulating mononuclear cells in GBS, as well as an important role for ?M-integrin-ICAM-1 interactions in pathogenic GBS patient leukocyte trafficking at the human BNB in vitro. PMID:22552879

  2. Polymorphonuclear leucocyte function in Behçet's disease

    Microsoft Academic Search

    J D Sobel; S Haim; N Obedeanu; T Meshulam; D Merzbach

    1977-01-01

    Polymorphonuclear leucocyte function was investigated in 19 patients with active Behçet's disease. Spontaneous free leucocyte migration was found to be significantly reduced, yet after stimulation the leucocyte's chemotactic activity was considerably increase (p less than 0-05) when compared to control leucocytes. Control leucocytes migrated more rapidly when incubated in serum taken from patients with Behçet's disease (p less than 0-005).

  3. Development of leukocyte cell lines from the channel catfish ( Ictalurus punctatus )

    Microsoft Academic Search

    Norman W. Miller; V. Gregory Chinchar; L. William Clem

    1994-01-01

    Summary Techniques are described for the generation of channel catfish long term leukocyte cell lines. These techniques include the isolation of peripheral blood leukocytes, purification of B cells by anti-immunoglobulin panning, mitogen stimulation, and in vitro maintenance and cloning of leukocyte cultures. Once stimulated in vitro, channel catfish leukocytes proliferate continuously without the need for exogenous growth factors or feeder

  4. Interactions through L-selectin between leukocytes and adherent leukocytes nucleate rolling adhesions on selectins and VCAM-1 in shear flow

    PubMed Central

    1996-01-01

    We demonstrate an additional step and a positive feedback loop in leukocyte accumulation on inflamed endothelium. Leukocytes in shear flow bind to adherent leukocytes through L-selectin/ligand interactions and subsequently bind downstream and roll on inflamed endothelium, purified E-selectin, P-selectin, L-selectin, VCAM-1, or peripheral node addressin. Thus adherent leukocytes nucleate formation of strings of rolling cells and synergistically enhance leukocyte accumulation. Neutrophils, monocytes, and activated T cell lines, but not peripheral blood T lymphocytes, tether to each other through L-selectin. L- selectin is not involved in direct binding to either E- or P-selectin and is not a major counterreceptor of endothelial selectins. Leukocyte- leukocyte tethers are more tolerant to high shear than direct tethers to endothelial selectins and, like other L-selectin-mediated interactions, require a shear threshold. Synergism between leukocyte- leukocyte and leukocyte-endothelial interactions introduces novel regulatory mechanisms in recruitment of leukocytes in inflammation. PMID:8909556

  5. Surface modification of polymeric materials and its effect on blood compatibility

    SciTech Connect

    Wrobleski, D.A.; Cash, D.L.; Archuleta, T.; Barthell, B.L.; Kossowsky, R.; London, J.E.; Lehnert, B.E.; Duchane, D.V.

    1987-01-01

    The surfaces of commercially available polymeric materials have been modified through the chemical infusion process and physical vapor deposition. The surfaces of poly(methylmethacrylate) (PMMA) have been modified through a chemical infusion process by treatment of the sample with a solution containing varying amounts of titanium(IV)isopropoxide and polyvinylpyrrolidone (PVP). The surfaces of silicone rubber samples have been coated with a thin coating of titanium dioxide with an ion beam sputtering technique. The treated samples were characterized by scanning electron microscopy, optical microscopy, and neutron activation analysis. The infused samples were evaluated for blood compatibility using two biological assays: an adherence assay in which the adherence of human polymorphonuclear leukocytes to the samples was determined, and a hemolysis assay using rat blood erythrocytes to determine the hemolytic activity of the samples. Based on the results of these assays, the PMMA samples treated with PVP alone resulted in an improvement in reactivity with the blood cells. 16 refs., 4 figs.

  6. Substitution of Aspartate for glycine 1018 in the Type III procollagen (COL3AI) gene causes type IV Ehlers-Danlos Syndrome: The mutated allele is present in most blood leukocytes of the asymptomatic and mosaic mother

    SciTech Connect

    Kontusaari, S.; Tromp, G.; Kuivaniemi, H.; Prockop, D.J. (Thomas Jefferson Univ., Philadelphia, PA (United States)); Stolle, C. (Robert Wood Johnson Medical School, Piscataway, NJ (United States)); Pope, F.M.

    1992-09-01

    A proband with arterial ruptures and skin changes characteristic of the type IV variant of Ehlers-Danlos syndrome was found to have a single-base mutation in the type III procollagen gene, which converted the codon for glycine at amino position 1018 to a codon for aspartate. (Amino acid positions are numbered by the standard convention in which the first glycine of the triple-helical domain of an [alpha] chain is number 1. The numbers of positions in the [alpha]1(III) chains can be converted to positions in the human pro[alpha](III) chain by adding 167.). Nucleotide sequencing of overlapping PCR products in which the two alleles were distinguished demonstrated that the mutation of glycine 1018 was the only mutation that changed the primary structure of type III procollagen. The glycine substitution markedly decreased the amount of type III procollagen secreted into the medium by cultured skin fibroblasts from the proband. It is surprising that the same mutation was found in about 94% of the peripheral blood leukocytes from the proband's asymptomatic 72-year-old mother. Other tissues from the mother contained the mutated allele; it was present in 0%-100% of different samples of hair cells and in about 40% of cells from the oral epithelium. Therefore, the mother was a mosaic for the mutation. Since the mutated allele was present in cells derived from all three germ layers, the results indicated that the mutation arose by the late blastocyst stage of development. The results also indicate that assays of blood leukocytes do not always reveal mosaicism or predict phenotypic involvement of tissues, such as blood vessels, that are derived from the same embryonic cells as are leukocytes. 66 refs., 6 figs., 1 tab.

  7. Multiple Heparanases Are Expressed in Polymorphonuclear Cells

    Microsoft Academic Search

    Mary Ann Kosir; Patricia A. Foley-Loudon; Raphaela Finkenauer; Steven D. Tennenberg

    2002-01-01

    Background. Heparan sulfate proteoglycans are complex cell surface molecules containing polysaccharides called heparan sulfate. Lysosomes, platelet granules, and neutrophils (polymorphonuclear cells) contain heparanases that degrade heparan sulfate. There are at least two groups of heparanases: connective tissue–activating-peptide (CTAP-III) and mammalian heparanase (hpa). The purpose of this study was to quantify the expression of both CTAP-III and hpa in neutrophils and

  8. Severity of localized juvenile periodontitis as related to polymorphonuclear chemotaxis and specific microbial isolates.

    PubMed

    Chinwalla, J; Tosi, M; Bissada, N F

    1998-01-01

    The purpose of this study was twofold: (a) to determine if an association exists between severity of localized juvenile periodontitis (LJP) and impairment of polymorphonuclear leukocyte (PMN) chemotaxis and/or colonization by specific microbial isolates; (b) to determine if the number of specific microbial isolates, i.e., Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, Eikenella corrodens, and Campylobacter rectus correlates to clinical severity of sites with LJP. Thirty-six first molars in nine subjects with LJP were examined. A clinical severity score was computed based on attachment level and vertical bone loss. The mean score of the four sites was designated as the clinical severity score for each subject. A DNA probe was used to quantitate colonization by A. actinomycetemcomitans, P. gingivalis, P. intermedia, F. nucleatum, E. corrodens, and C. rectus. The chemotaxis of isolated peripheral blood PMNs was measured in Boyden chambers using the leading front technique. It was found that the clinical severity score ranged from 7.8 for mild involvement to 32.5 for severe LJP. The chemotaxis result for each patient (micron/90 min) was expressed as a percentage of the value for a paired healthy control and ranged from 40% to 104%. Four of the nine subjects had chemotaxis values less than 70% of control. Linear regression analysis showed: (a) no significant correlation between PMN chemotaxis and severity of LJP in subjects (r = .14); (b) a significant correlation (r = 0.43) between severe sites with LJP and number of specific microbial isolates. Sites with severe LJP had present at least five different microbial isolates as compared with non-severe sites (P < .05). No particular trend of colonization by periodontopathogens was seen in non-severe sites. Further, severe sites of LJP displayed a statistically significant greater colonization by P. intermedia, E. corrodens, and C. rectus than did non-severe sites (P < .05). It may be concluded that neither the degree of PMN chemotactic impairment nor A. actinomycetemcomitans is a reliable indicator of the severity of LJP. PMID:9663117

  9. Measurement of interferon-tau (IFN-?) stimulated gene expression in blood leukocytes for pregnancy diagnosis within 18–20 d after insemination in dairy cattle

    Microsoft Academic Search

    J. C. Green; C. S. Okamura; S. E. Poock; M. C. Lucy

    2010-01-01

    The objective was to diagnose pregnancy within 18d after insemination by measuring interferon-tau (IFN-?)-stimulated genes (ISG) expression in circulatory leukocytes. Based on microarray results, three genes were selected [2?-5? oligoadenylate synthetase 1(Oas1), myxovirus resistance gene 2 (Mx2), and interferon-stimulated gene 15kDa protein (Isg15)] because they were known to be interferon-stimulated genes (ISG) and were also differentially expressed in the samples.

  10. Human leukocyte antigen (HLA)-G during pregnancy part I: correlations between maternal soluble HLA-G at midterm, at term, and umbilical cord blood soluble HLA-G at term.

    PubMed

    Klitkou, Louise; Dahl, Mette; Hviid, Thomas Vauvert F; Djurisic, Snezana; Piosik, Zofia Maria; Skovbo, Peter; Mřller, Anna Margrethe; Steffensen, Rudi; Christiansen, Ole B

    2015-04-01

    Human leukocyte antigen (HLA)-G is a class Ib molecule with restricted tissue distribution expressed on trophoblast cells and has been proposed to have immunomodulatory functions during pregnancy. Soluble HLA-G1 (sHLA-G1) can be generated by the shedding of membrane-bound HLA-G molecules; however, three soluble isoforms also exist (HLA-G5 to -G6). During pregnancy, it is unknown whether there is a correlation between sHLA-G levels in maternal and fetal blood. In 246 pregnancies, we have measured the levels of sHLA-G1/-G5 in maternal blood plasma samples from gestational week 20 (GW20) and at term, as well as in umbilical cord blood samples. Soluble HLA-G levels declined by 38.4% in maternal blood from GW20 to term, and sHLA-G levels were significantly lower in maternal blood at term than in GW20 (P<0.001). At term, the sHLA-G levels were significantly higher in maternal blood than in umbilical blood (P<0.001). HLA-G levels in maternal blood in GW20 and at term, and in maternal blood at term and umbilical cord blood, were correlated (P<0.001 and P<0.01, respectively). This is the first large study simultaneously measuring sHLA-G in both maternal and umbilical cord blood. The finding that sHLA-G levels are significantly lower in fetal compared with maternal blood at term documents for the first time that sHLA-G is not freely transferred over the placental barrier. Soluble HLA-G levels in maternal and fetal blood were found to be correlated, which may be due to shared genetic factors of importance for production of sHLA-G in the mother and child, or it may support the theory that sHLA-G in the pregnant woman and the fetus is partly derived from a "shared organ", the placenta. PMID:25636573

  11. Effects of phorbol myristate acetate-stimulated human leukocytes on rat lung.

    PubMed

    Perry, M L; Kayes, S G; Barnard, J W; Taylor, A E

    1990-01-01

    Human blood was separated into polymorphonuclear (PMN) and mononuclear (MN) leukocyte fractions, and 3 x 10(7) cells (PMN or MN) were added to isolated rat lungs perfused with 5% human albumin in buffer and stimulated with phorbol myristate acetate (PMA). Lungs perfused with either albumin alone, PMN, or MN but not stimulated with PMA showed no change in vascular resistance or endothelial permeability measured as the capillary filtration coefficient (Kf,c). Lungs that were stimulated with PMA with no cells showed no change in Kf,c (0.34 +/- 0.07 vs. 0.37 +/- 0.7), but vascular resistance increased in all segments of the circulation. Capillary pressure, the major force responsible for edema formation, nearly doubled in the absence of cells 40 min after PMA. Lungs perfused with either PMN or MN and stimulated with PMA were injured. Kf,c increased from 0.41 +/- 0.03 to 0.87 +/- 0.10 (PMN) and from 0.36 +/- 0.07 to 0.81 +/- 0.23 (MN) 90 min after PMA. In addition to the increased endothelial permeability, vascular resistances and pressures also increased in the cell-perfused PMA-stimulated lungs. These results demonstrate that cells other than granulocytes are capable of producing severe acute lung injury and cannot be ignored when the effects of PMA on neutrophil-depleted lungs are studied. PMID:2312463

  12. [Adaptability of Opisthorchis felineus metacercariae as a result of altered phagocytic activity of polymorphonuclear neutrophils].

    PubMed

    Iakovleva, V V; Stepanova, T F; Sozonova, T A; Rybakova, M V; Parfenova, G V

    2002-01-01

    The higher absorptivity of neutrophils during their alpha-tocopherol stimulation has been found to promote lower adaptability in the metacercariae of Opisthorchis felineus, a reverse picture was observed when cyclophosphanum was used. The antihelminthic effect of biltricide is probably to be unassociated with the changed functional activity of polymorphonuclear leukocytes, but to be determined by its direct action on helminths. This phenomenon was studied on golden hamsters inoculated with Opisthorchis felineus. On day 2 after inoculation, the following drugs were given for a fortnight: biltricide in a single dose of 60 mg/kg weight; biltricide used in a daily fractionated dose of 60 mg/kg; cyclophosphanum (6 mg/kg, every second day); alpha-tocopherol in a daily dose of 30 mg every day. PMID:12224258

  13. Hematologically important mutations: Leukocyte adhesion deficiency (first update)

    Microsoft Academic Search

    Edith van de Vijver; Anne Maddalena; Özden Sanal; Steven M. Holland; Gulbu Uzel; Manisha Madkaikar; Martin de Boer; Karin van Leeuwen; M. Yavuz Köker; Nima Parvaneh; Alain Fischer; S. K. Alex Law; Nigel Klein; F. Ilhan Tezcan; Ekrem Unal; Turkan Patiroglu; Bernd Belohradsky; Klaus Schwartz; Raz Somech; Taco W. Kuijpers; Dirk Roos

    Leukocyte adhesion deficiency (LAD) is an immunodeficiency caused by defects in the adhesion of leukocytes (especially neutrophils) to the blood vessel wall. As a result, patients with LAD suffer from severe bacterial infections and impaired wound healing, accompanied by neutrophilia. In LAD-I, mutations are found in ITGB2, the gene that encodes the ? subunit of the ?2 integrins. This syndrome

  14. Coupled Flow-Structure-Biochemistry Simulations of Dynamic Systems of Blood Cells Using an Adaptive Surface Tracking Method

    PubMed Central

    Hoskins, M.H.; Kunz, R.F.; Bistline, J.E.; Dong, C.

    2009-01-01

    A method for the computation of low Reynolds number dynamic blood cell systems is presented. The specific system of interest here is interaction between cancer cells and white blood cells in an experimental flow system. Fluid dynamics, structural mechanics, six-degree-of freedom motion control and surface biochemistry analysis components are coupled in the context of adaptive octree-based grid generation. Analytical and numerical verification of the quasi-steady assumption for the fluid mechanics is presented. The capabilities of the technique are demonstrated by presenting several three-dimensional cell system simulations, including the collision/interaction between a cancer cell and an endothelium adherent polymorphonuclear leukocyte (PMN) cell in a shear flow. PMID:20160939

  15. Journal of Leukocyte Biology

    NSDL National Science Digital Library

    The Society for Leukocyte Biology, in conjunction with Stanford University's HighWire Press, has placed online most issues of the Journal of Leukocyte Biology. Devoted to "the exploration of the cellular and molecular biology of leukocytes," the journal is currently free "for a limited time" to all users (registration required). Full online text [.pdf] is available for all issues of 2000 and 2001; abstracts are available from 1984 onwards.

  16. Fast leukocyte image segmentation using shadowed sets.

    PubMed

    Mohapatra, Subrajeet; Patra, Dipti; Kumar, Kundan

    2012-01-01

    Leukocyte image segmentation acts as the foundation for all automated image based hematological disease recognition systems. Perfection in image segmentation is a necessary condition for improving the diagnostic accuracy in automated cytology. Even though much effort has been put in developing suitable segmentation routines, the problem still remains open in areas like pathological imaging. Clustering is an essential image segmentation procedure which segments an image into desired regions. This paper introduces a novel Shadowed C-means (SCM) clustering approach towards leukocyte segmentation in blood microscopic images. The segmented nucleus and cytoplasm of a leukocyte can be used for feature extraction which can lead to acute leukemia detection. Absence of parameter tuning in SCM with acceptable segmentation performance gives the proposed scheme an edge over standard cluster based segmentation techniques. Comparative analysis reveals that the proposed algorithm is fast and robust in segmenting stained blood microscopic images in the presence of outliers. PMID:22436298

  17. Three or more routes for leukocyte migration into the central nervous system

    Microsoft Academic Search

    Pia Kivisäkk; Grahame Kidd; Richard M. Ransohoff

    2003-01-01

    Leukocyte migration into and through tissues is fundamental to normal physiology, immunopathology and host defence. Leukocyte entry into the central nervous system (CNS) is restricted, in part, because of the blood–brain barrier (BBB). During the past decade, crucial components that are involved in the process of leukocyte migration have been identified and progress has been made in understanding the mechanisms

  18. Leukocyte depletion results in improved lung function and reduced inflammatory response after cardiac surgery

    Microsoft Academic Search

    Y. J. Gu; A. J. deVries; P. W. Boonstra; W. van Oeveren

    1996-01-01

    Leukocyte depletion during cardiopulmonary bypass has been demonstrated in animal experiments to improve pulmonary function. Conflicting results have been reported, however, with clinical depletion by arterial line filter of leukocytes at the beginning of cardiopulmonary bypass. In this study, we examined whether leukocyte depletion from the residual heart-lung machine blood at the end of cardiopulmonary bypass would improve lung function

  19. Blood

    MedlinePLUS

    ... a mixture of blood cells and plasma. Continue Red Blood Cells Red blood cells (RBCs, and also ... conditions involving the blood include: Diseases of the Red Blood Cells The most common condition affecting the ...

  20. Oxygen radical production by avian leukocytes.

    PubMed Central

    Conlon, P; Smith, D; Gowlett, T

    1991-01-01

    Oxygen radical production by heterophils of red-tailed hawks and chickens, and by neutrophils of calves, was evaluated in a chemiluminescence microassay. Leukocytes were isolated by centrifugation of blood in capillary tubes and then challenged with opsonized zymosan in the presence of luminol. Avian heterophils produced significantly fewer oxygen radicals than did bovine neutrophils. PMID:1884301

  1. Analysis of blood leukocytes in a naturally occurring immunodeficiency of pigs shows the defect is localized to B and T cells.

    PubMed

    Ewen, C L; Cino-Ozuna, A G; He, H; Kerrigan, M A; Dekkers, J C M; Tuggle, C K; Rowland, R R R; Wyatt, C R

    2014-12-15

    Severe combined immunodeficiency (SCID) is the result of a set of inherited genetic defects which render components of the immune response nonfunctional. In Arabian horses, Jack Russell terriers, and mice, the disorder is a consequence of the absence of T and B lymphocytes, while natural killer (NK) cell and other leukocyte populations remain intact. Preliminary analysis of a naturally acquired form of inherited SCID in a line of pigs showed several defects in the architecture and composition of secondary lymphoid organs. In this study, a quantitative assessment of lymphocyte populations in affected and normal littermates showed depleted T or B lymphocyte populations in affected pigs; however, NK cells and neutrophils were present in numbers comparable to unaffected littermates. The results indicate that the immune defect in pigs shares the same features as other SCID-affected species. PMID:25454085

  2. Endometrial leukocytes and menstruation

    Microsoft Academic Search

    Lois A. Salamonsen; Louise J. Lathbury

    2000-01-01

    This review examines evidence supporting the concept that menstruation occurs as a result of an inflammatory process. In the endometrium, leukocyte numbers rise in the late secretory phase following the fall in serum progesterone concentrations. It is postulated that products released following activation of these leukocytes are critically important for menstruation. Mast cells, eosinophils, neutrophils and macrophages in particular are

  3. Monitoring of benzene-induced hematotoxicity in mice by serial leukocyte counting using a microcavity array.

    PubMed

    Hosokawa, Masahito; Asami, Marie; Yoshino, Tomoko; Tsujimura, Noriyuki; Takahashi, Masayuki; Nakasono, Satoshi; Tanaka, Tsuyoshi; Matsunaga, Tadashi

    2013-02-15

    Monitoring of hematotoxicity, which requires serial blood collection, is difficult to carry out in small animals due to a lack of non-invasive, individual animal-appropriate techniques that enable enumeration of leukocyte subsets from limited amounts of whole blood. In this study, a microfluidic device equipped with a microcavity array that enables highly efficient separation of leukocytes from submicroliters of whole blood was applied for hematotoxicity monitoring in mice. The microcavity array can specifically separate leukocytes from whole blood based on differences in the size and deformability between leukocytes and other blood cells. Mouse leukocytes recovered on aligned microcavities were continuously processed for image-based immunophenotypic analysis. Our device successfully recovered almost 100% of mouse leukocytes in 0.1 ?L of whole blood without the effect of serial blood collection such as changes in body weight and total leukocyte count. We assessed benzene-associated hematotoxicity in mice using this system. Mice were administered with benzene once daily and the depression of leukocyte numbers induced in individual mice was successfully monitored from tail vein blood collected every other day for 2 weeks. Serial monitoring of the leukocyte number in individual mice will contribute to the understanding of hematotoxicity and reduction of the number of animal experiment trials. PMID:22770906

  4. The effects of cannabinoids and cannabispiro compounds on Escherichia coli adhesion to tissue culture cells and on leukocyte functions in vitro.

    PubMed

    Molnár, J; Petri, I; Berek, I; Shoyama, Y; Nishioka, I

    1987-01-01

    delta 9-Tetrahydrocannabinol, cannabidiol, cannabidiolic acid, tetrahydrocannabidiolic acid, cannabispirol, acetylcannabispirol, cannabispirone, and cannabispirenone in a low concentration did not affect the adhesion of Escherichia coli on cultured HEp-2 cells. Cannabinoids at 10(-6) M increased the chemiluminescence of human polymorphonuclear leukocytes, while the cannabispiro compounds failed to enhance the oxidative burst of leukocytes. In lymphocyte and granulocyte function tests (E- and EA-rosette formation, blast transformation of T-lymphocytes in the presence of phytohaemagglutinin and concanavalin-A, ADCC and phagocytosis) all compounds displayed immunosuppressive effect at 1.5 X 10(-5) M. Tetrahydrocannabidiolic acid exerted the weakest immunosuppression on human leukocyte functions. PMID:3329437

  5. Lipoxin A4 and aspirin-triggered 15-epi-lipoxin A4 inhibit peroxynitrite formation, NF-?B and AP-1 activation, and IL-8 gene expression in human leukocytes

    PubMed Central

    József, Levente; Zouki, Christine; Petasis, Nicos A.; Serhan, Charles N.; Filep, János G.

    2002-01-01

    Lipoxin A4 (LXA4) and aspirin-triggered 15-epi-LXA4 (ATL) are emerging as endogenous braking signals for neutrophil-mediated tissue injury. Recent studies indicate that peroxynitrite (ONOO?) may function as an intracellular signal for the production of IL-8, a potent proinflammatory cytokine in human leukocytes. In this study, we evaluated the impact of the metabolically stable analogues of LXA4/ATL on lipopolysaccharide (LPS)-induced ONOO? formation and ONOO?-mediated IL-8 gene expression in human leukocytes. At nanomolar concentrations, LXA4 analogues markedly reduced LPS-stimulated superoxide formation, evoked increases in intracellular diamino-fluorescein fluorescence (an indicator of NO formation), and consequently reduced ONOO? formation in isolated neutrophils, as well as in neutrophils, monocytes, and lymphocytes, in whole blood. LXA4/ATL analogues attenuated nuclear accumulation of activator protein-1 and nuclear factor-?B in both polymorphonuclear and mononuclear leukocytes and inhibited IL-8 mRNA expression and IL-8 release by 50–65% in response to LPS. The LXA4 inhibitory responses were concentration dependent and were not shared by 15-deoxy-LXA4. None of the LXA4 analogues studied affected neutrophil survival, nor reversed the apoptosis delaying action of LPS in neutrophils. In addition, LXA4 analogues had no significant effect on exogenous ONOO?-induced IL-8 gene and protein expression. These findings suggest that by attenuating ONOO? formation, LXA4 and ATL can oppose ONOO? signaling in leukocytes and provide a rationale for using stable synthetic analogues as antiinflammatory compounds in vivo. PMID:12235371

  6. Endogenous Tetrapyrroles Influence Leukocyte Responses to Lipopolysaccharide in Human Blood: Pre-Clinical Evidence Demonstrating the Anti-Inflammatory Potential of Biliverdin.

    PubMed

    Bisht, Kavita; Tampe, Jens; Shing, Cecilia; Bakrania, Bhavisha; Winearls, James; Fraser, John; Wagner, Karl-Heinz; Bulmer, Andrew C

    2014-05-30

    Sepsis is associated with abnormal host immune function in response to pathogen exposure, including endotoxin (lipopolysaccharide; LPS). Cytokines play crucial roles in the induction and resolution of inflammation in sepsis. Therefore, the primary aim of this study was to investigate the effects of endogenous tetrapyrroles, including biliverdin (BV) and unconjugated bilirubin (UCB) on LPS-induced cytokines in human blood. Biliverdin and UCB are by products of haem catabolism and have strong cytoprotective, antioxidant and anti-inflammatory effects. In the present study, whole human blood supplemented with BV and without was incubated in the presence or absence of LPS for 4 and 8 hours. Thereafter, whole blood was analysed for gene and protein expression of cytokines, including IL-1?, IL-6, TNF, IFN-?, IL-1Ra and IL-8. Biliverdin (50 ?M) significantly decreased the LPS-mediated gene expression of IL-1?, IL-6, IFN-?, IL-1Ra and IL-8 (P<0.05). Furthermore, BV significantly decreased LPS-induced secretion of IL-1? and IL-8 (P<0.05). Serum samples from human subjects and, wild type and hyperbilirubinaemic Gunn rats were also used to assess the relationship between circulating bilirubin and cytokine expression/production. Significant positive correlations between baseline UCB concentrations in human blood and LPS-mediated gene expression of IL-1? (R=0.929), IFN-? (R=0.809), IL-1Ra (R=0.786) and IL-8 (R=0.857) were observed in blood samples (all P<0.05). These data were supported by increased baseline IL-1? concentrations in hyperbilirubinaemic Gunn rats (P<0.05). Blood samples were also investigated for complement receptor-5 (C5aR) expression. Stimulation of blood with LPS decreased gene expression of C5aR (P<0.05). Treatment of blood with BV alone and in the presence of LPS tended to decrease C5aR expression (P=0.08). These data indicate that supplemented BV inhibits the ex vivo response of human blood to LPS. Surprisingly, however, baseline UCB was associated with heighted inflammatory response to LPS. This is the first study to explore the effects of BV in a preclinical human model of inflammation and suggests that BV could represent an anti-inflammatory target for the prevention of LPS mediated inflammation in vivo. PMID:25177524

  7. THE BOVINE NEUTROPHIL: STRUCTURE AND FUNCTION IN BLOOD AND MILK

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Polymorphonuclear neutrophil leukocytes (PMN) form the first line of cellular defense against invading pathogens. The PMN is characterized by a polymorphic segmented nucleus, numerous cytoplasmic granules that provide constituents for killing bacteria, large stores of glycogen for energy and a high...

  8. Effects of the Tumor-Leukocyte Microenvironment on Melanoma–Neutrophil Adhesion to the Endothelium in a Shear Flow

    PubMed Central

    Liang, Shile; Hoskins, Meghan; Khanna, Payal; Kunz, Robert F.; Dong, Cheng

    2009-01-01

    The primary cause of cancer mortality is not attributed to primary tumor formation, but rather to the growth of metastases at distant organ sites. Tumor cell adhesion to blood vessel endothelium (EC) and subsequent transendothelial migration within the circulation are critical components of the metastasis cascade. Previous studies have shown polymorphonuclear neutrophils (PMNs) may facilitate melanoma cell adhesion to the EC and subsequent extravasation under flow conditions. The melanoma cell–PMN interactions are found to be mediated by the binding between intercellular adhesion molecule-1 (ICAM-1) on melanoma cells and ?2 integrin on PMNs and by endogenously secreted interleukin 8 (IL-8) within the tumor-leukocyte microenvironment. In this study, the effects of fluid convection on the IL-8-mediated activation of PMNs and the binding kinetics between PMNs and melanoma cells were investigated. Results indicate that the shear rate dependence of PMN–melanoma cell adhesion and melanoma cell extravasation is due, at least partly, to the convection of tumor-secreted proinflammatory cytokine IL-8. PMID:19865613

  9. Defective polymorphonuclear leukocyte formyl peptide receptor(s) in juvenile periodontitis.

    PubMed

    Perez, H D; Kelly, E; Elfman, F; Armitage, G; Winkler, J

    1991-03-01

    Juvenile periodontitis (JP) is a disease characterized by severe gingival infections. PMN from some JP patients exhibit abnormal chemotactic responsiveness when challenged with the synthetic formyl peptide, FMLP. While investigating PMN function in JP, we found a patient in whom abnormal PMN chemotactic responses to FMLP were associated with a defective population of PMN formyl peptide receptor(s) (FPR). JP PMN failed to respond chemotactically when challenged with FMLP, but exhibited normal chemotactic responses upon exposure to purified human C5a. Furthermore, JP PMN were capable of degranulating and generating superoxide anion radicals as well as normal PMN upon exposure to FMLP. Binding studies demonstrated that JP PMN had a diminution in the number of high-affinity FPR. Studies in which FPR was radiolabeled by chemical cross-linking demonstrated that JP PMN FPR exhibited the same molecular weight and N-linked glycosylation as normal PMN FPR. JP PMN FPR, however, was more resistant to papain cleavage than normal PMN FPR. Autoradiograms obtained from 2D-PAGE of normal and JP PMN FPR demonstrated decreased amounts of FPR isoforms in JP PMN. PMID:1999504

  10. Defective polymorphonuclear leukocyte formyl peptide receptor(s) in juvenile periodontitis.

    PubMed Central

    Perez, H D; Kelly, E; Elfman, F; Armitage, G; Winkler, J

    1991-01-01

    Juvenile periodontitis (JP) is a disease characterized by severe gingival infections. PMN from some JP patients exhibit abnormal chemotactic responsiveness when challenged with the synthetic formyl peptide, FMLP. While investigating PMN function in JP, we found a patient in whom abnormal PMN chemotactic responses to FMLP were associated with a defective population of PMN formyl peptide receptor(s) (FPR). JP PMN failed to respond chemotactically when challenged with FMLP, but exhibited normal chemotactic responses upon exposure to purified human C5a. Furthermore, JP PMN were capable of degranulating and generating superoxide anion radicals as well as normal PMN upon exposure to FMLP. Binding studies demonstrated that JP PMN had a diminution in the number of high-affinity FPR. Studies in which FPR was radiolabeled by chemical cross-linking demonstrated that JP PMN FPR exhibited the same molecular weight and N-linked glycosylation as normal PMN FPR. JP PMN FPR, however, was more resistant to papain cleavage than normal PMN FPR. Autoradiograms obtained from 2D-PAGE of normal and JP PMN FPR demonstrated decreased amounts of FPR isoforms in JP PMN. Images PMID:1999504

  11. Factors affecting ability of skim milk to support phagocytosis by bovine polymorphonuclear leukocytes.

    PubMed

    Miller, R H; Paape, M J; Dulin, A M; Schultze, W D; Weinland, B T; Pearson, R E

    1985-11-01

    Milk samples from quarters were collected from 48 Holstein cows at 1.5, 3, 21, and 35 wk postpartum and at 1 wk after drying off. Foremilk samples were obtained for bacteriological examination, somatic cell count of milk, and for assay of phagocytosis. Thirty-two cows in first lactation and 16 in third lactation produced by two criteria of sire selection (high milk versus multiple trait) were studied. In vitro phagocytosis was assayed four times on each original quarter milk sample, yielding 3,840 determinations. Variance components were large for cow-associated variation. Cow, stage X cow, and cow X sample dilution were 18.7, 20.6, and 6.0% of total variance. Phagocytosis was 9% higher in skim milk from third lactations than from first. Phagocytosis was highest in dry period samples, followed by 1.5, 35, 3, and 21-wk postpartum samples. Milk somatic cell count tended to be related more closely to phagocytosis than was current bacteriological status of the quarter. Skim milk from genetically higher producing cows was less conducive to phagocytosis than skim milk from genetically lower producing cows, possibly because of dilution. PMID:4078134

  12. Age-related comparison of superoxide production by canine neutrophilic polymorphonuclear leukocytes 

    E-print Network

    Hanson, Thomas Dale

    1983-01-01

    adequate I diet under conventional conditions in outdoor pens. Many of the aged dogs suffered from various infirmities considered typical of the types of maladies affecting aged mammals. The young beagles were free of any obvious disease. Leukoc te... reported have been shown in humans or in animals with inherited anomalies and/or pathological conditions, i. e. CGS in dogs and CGD in humans (37, 41, 52, 65-67). Reports of age-related differences in PMN oxidative capacity are few. A decrease...

  13. Effects of 60 Hz magnetic field exposure on polymorphonuclear leukocyte activation

    Microsoft Academic Search

    Raddassi Khadir; James L Morgan; John J Murray

    1999-01-01

    We have investigated the effects of a sinusoidal 60 Hz magnetic field on free radical (superoxide anion) production, degranulation (?-glucuronidase and lysozyme release) and viability in human neutrophils (PMNs). Experiments were performed blindly in very controlled conditions to examine the effects of a magnetic field in resting PMNs and in PMNs stimulated with a tumor promoter: phorbol 12-myristate 13-acetate (PMA).

  14. Effects of 60 Hz magnetic field exposure on polymorphonuclear leukocyte activation.

    PubMed

    Khadir, R; Morgan, J L; Murray, J J

    1999-10-18

    We have investigated the effects of a sinusoidal 60 Hz magnetic field on free radical (superoxide anion) production, degranulation (beta-glucuronidase and lysozyme release) and viability in human neutrophils (PMNs). Experiments were performed blindly in very controlled conditions to examine the effects of a magnetic field in resting PMNs and in PMNs stimulated with a tumor promoter: phorbol 12-myristate 13-acetate (PMA). Exposure of unstimulated human PMNs to a 60 Hz magnetic field did not affect the functions examined. In contrast, exposure of PMNs to a 22 milliTesla (mT), 60 Hz magnetic field induced significant increases in superoxide anion (O2-) production (26.5%) and in beta-glucuronidase release (53%) when the cells were incubated with a suboptimal stimulating dose of PMA. Release of lysozyme and lactate dehydrogenase was unchanged by the magnetic field, whether the cells were stimulated or not. A 60 Hz magnetic field did not have any effect on O2- generation by a cell-free system xanthine/xanthine oxidase, suggesting that a magnetic field could upregulate common cellular events (signal transduction) leading to O2- generation and beta-glucuronidase release. In conclusion, exposure of PMNs to a 22 mT, 60 Hz magnetic field potentiates the effect of PMA on O2- generation and beta-glucuronidase release. This effect could be the result of an alteration in the intracellular signaling. PMID:10572957

  15. Fate of surface proteins of rabbit polymorphonuclear leukocytes during phagocytosis. I. Identification of surface proteins

    PubMed Central

    1979-01-01

    To study the fate of external membrane proteins during phagocytosis, rabbit peritoneal neutrophils were labeled by enzymatic iodination. Iodine was incorporated into at least 13 proteins ranging in size from approximately 250,000 to 18,000 daltons as judged from autoradiography of gels after SDS-polyacrylamide gel electrophoresis of labeled cells. The major contractile proteins of neutrophils, actin and myosin, were not labeled when intact cells were iodinated but were labeled when homogenates of these cells were iodinated. Nine of the iodinated proteins were released by mild protease treatment of intact cells. A plasma membrane-rich fraction was isolated by density centrifugation. This fraction was enriched at least 10-fold for lactoperoxidase-labeled acid-insoluble proteins. It was enriched to the same extent for the presence of iodinated wheat germ agglutinin that had been bound to intact cells at 4 degrees C before homogenization. Analysis of SDS- polyacrylamide gel electrophoresis revealed that the proteins of this fraction were predominantly of high molecular weight. However, only 8 of the 13 proteins iodinated on intact cells were found in this fraction. The remaining five were enriched in a dense fraction containing nuclei, intact cells, and membranous vesicles, and may represent a specialized segment of the neutrophil cell surface. PMID:479301

  16. Novel post-translational incorporation of tyrosine in PMA-activated polymorphonuclear leukocytes (PMN)

    SciTech Connect

    Nath, J.; Oliver, C.; Ohno, Y.; Gallin, J.I.

    1986-03-05

    During studies undertaken to determine whether stimulation of tubulin tyrosinolation occurs in PMA-activated PMN, a distinctly different and novel post-translational incorporation of tyrosine into multiple PMN proteins was observed. The reaction also occurred in organelle-depleted neutrophil cytoplasts and was highly exaggerated in organelle-enriched karyogranuloplasts. The incorporation was specific for tyrosine, did not require extracellular Ca/sup 2 +/ and was inhibited in the presence of a variety of reducing agents, intracellular scavengers of oxygen radicals and inhibitors of peroxidase-mediated reactions. The PMA-induced incorporation of tyrosine was completely absent in PMN from patients with chronic granulomatous disease, but occurred normally in PMN of a patient with myeloperoxidase deficiency. Moreover, the incorporation of tyrosine was blocked by N-acetyl-L-tyrosine but not by phenylalanine suggesting a requirement for the phenolic group. A two-fold increase in stable protein carbonyl derivatives was demonstrated suggesting an increased oxidative modification of the proteins. SDS urea PAGE and reversed phase HPLC did not reveal any detectable changes in the extent of protein cross-linking. The PMN tyrosine pool was approximately 900 ..mu..M and yet only 1 ..mu..M tyrosine was added in these experiments. The functional significance of this reaction is not yet clear.

  17. SERUM INHIBITION OF THE OXIDATIVE BURST IN HUMAN POLYMORPHONUCLEAR LEUKOCYTES. (R826781)

    EPA Science Inventory

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  18. SERUM INHIBITION OF THE OXIDATIVE BURST IN HUMAN POLYMORPHONUCLEAR LEUKOCYTES. (R827354C003)

    EPA Science Inventory

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  19. The Fruiting Bodies, Submerged Culture Biomass, and Acidic Polysaccharide Glucuronoxylomannan of Yellow Brain Mushroom Tremella mesenterica Modulate the Immunity of Peripheral Blood Leukocytes and Splenocytes in Rats with Impaired Glucose Tolerance.

    PubMed

    Hsu, Tai-Hao; Lee, Chien-Hsing; Lin, Fang-Yi; Wasser, Solomon P; Lo, Hui-Chen

    2014-01-01

    The prevalence of diabetes mellitus (DM), a chronic disease with hyperglycemia and impaired immune function, is increasing worldwide. Progression from impaired glucose tolerance (IGT) to type 2 DM has recently become a target for early intervention. The fruiting bodies (FB) and submerged culture mycelium (CM) of Tremella mesenterica, an edible and medicinal mushroom, have been demonstrated to have antihyperglycemic and immunomodulatory activities in type 1 DM rats. Herein, we investigated the effects of acidic polysaccharide glucuronoxylomannan (GX) extracted from CM on the immunocyte responses. Male Wistar rats were injected with streptozotocin (65 mg/kg) plus nicotinamide (200 mg/kg) for the induction of IGT, and gavaged daily with vehicle, FB, CM, or GX (1 g/kg/day). Rats injected with saline and gavaged vehicle were used as controls. Two weeks later, peripheral blood leukocytes (PBLs) and splenocytes were collected. Ingestion of FB, CM, and GX significantly decreased blood glucose levels in the postprandial period and in oral glucose tolerance test, and partially reversed T-splenocytic proliferation in IGT rats. CM significantly decreased T-helper lymphocytes in the PBLs and B-splenocytes. In addition, FB, CM, and GX significantly reversed the IGT-induced decreases in tumor necrosis factor-? production; GX significantly increased interleukin-6 production in T-lymphocytes in the PBLs and splenocytes; and CM and GX significantly reversed IGT-induced decrease in interferon-? production in T-lymphocytes in the spleen. In conclusion, FB, CM, and acidic polysaccharide GX of T. mesenterica may increase T-cell immunity via the elevation of proinflammatory and T-helper cytokine production in rats with impaired glucose tolerance. PMID:24872934

  20. The Fruiting Bodies, Submerged Culture Biomass, and Acidic Polysaccharide Glucuronoxylomannan of Yellow Brain Mushroom Tremella mesenterica Modulate the Immunity of Peripheral Blood Leukocytes and Splenocytes in Rats with Impaired Glucose Tolerance

    PubMed Central

    Hsu, Tai-Hao; Lee, Chien-Hsing; Lin, Fang-Yi; Wasser, Solomon P.; Lo, Hui-Chen

    2014-01-01

    The prevalence of diabetes mellitus (DM), a chronic disease with hyperglycemia and impaired immune function, is increasing worldwide. Progression from impaired glucose tolerance (IGT) to type 2 DM has recently become a target for early intervention. The fruiting bodies (FB) and submerged culture mycelium (CM) of Tremella mesenterica, an edible and medicinal mushroom, have been demonstrated to have antihyperglycemic and immunomodulatory activities in type 1 DM rats. Herein, we investigated the effects of acidic polysaccharide glucuronoxylomannan (GX) extracted from CM on the immunocyte responses. Male Wistar rats were injected with streptozotocin (65 mg/kg) plus nicotinamide (200 mg/kg) for the induction of IGT, and gavaged daily with vehicle, FB, CM, or GX (1 g/kg/day). Rats injected with saline and gavaged vehicle were used as controls. Two weeks later, peripheral blood leukocytes (PBLs) and splenocytes were collected. Ingestion of FB, CM, and GX significantly decreased blood glucose levels in the postprandial period and in oral glucose tolerance test, and partially reversed T-splenocytic proliferation in IGT rats. CM significantly decreased T-helper lymphocytes in the PBLs and B-splenocytes. In addition, FB, CM, and GX significantly reversed the IGT-induced decreases in tumor necrosis factor-? production; GX significantly increased interleukin-6 production in T-lymphocytes in the PBLs and splenocytes; and CM and GX significantly reversed IGT-induced decrease in interferon-? production in T-lymphocytes in the spleen. In conclusion, FB, CM, and acidic polysaccharide GX of T. mesenterica may increase T-cell immunity via the elevation of proinflammatory and T-helper cytokine production in rats with impaired glucose tolerance. PMID:24872934

  1. Nitration of tyrosyl-residues from extra- and intracellular proteins in human whole blood.

    PubMed

    Salman-Tabcheh, S; Guérin, M C; Torreilles, J

    1995-11-01

    We measured the amounts of tyrosine and 3-nitrotyrosine (NO2-tyrosine) in proteins of plasma and polymorphonuclear leukocytes (PMN) from human whole blood before and after activation with phorbol ester (PMA) or calcium ionophore (A 23187). In unstimulated blood, no significant nitration of tyrosine was detected into PMN proteins, but a NO2-tyrosine/tyrosine ratio of 0.7% was detected in plasma proteins. When blood was activated with PMA, the NO2-tyrosine/tyrosine ratio stayed at 0.7% in plasma proteins, but it increased to 1.4% in PMN proteins, indicating a peroxynitrite production within the cells. In blood activated with calcium ionophore, the NO2-tyrosine/tyrosine ratio was 1.2% in plasma proteins and 2.1% in PMN proteins. Incubation of blood with a NO-synthase inhibitor before stimulation inhibited such a protein tyrosine nitration. To ensure that NO2-tyrosine detected in intracellular proteins did not result from the enzymatic posttranslational tyrosylation of PMN proteins, the incorporation of 14C labeled tyrosine into PMN proteins after activation with PMA or A23187 was studied. The addition of a 10 fold excess of NO2-tyrosine did not modify the course of protein tyrosylation. Because tyrosine nitration is an irreversible reaction, NO2-tyrosine could be accumulated into proteins and could act as a cumulative index of peroxynitrite production. PMID:8529931

  2. C-type natriuretic peptide inhibits leukocyte recruitment and platelet-leukocyte interactions via suppression of P-selectin expression

    NASA Astrophysics Data System (ADS)

    Scotland, Ramona S.; Cohen, Marc; Foster, Paul; Lovell, Matthew; Mathur, Anthony; Ahluwalia, Amrita; Hobbs, Adrian J.

    2005-10-01

    The multifaceted process of immune cell recruitment to sites of tissue injury is key to the development of an inflammatory response and involved in the pathogenesis of numerous cardiovascular disorders. We recently identified C-type natriuretic peptide (CNP) as an important endothelium-derived mediator that regulates vascular tone and protects against myocardial ischemia/reperfusion injury. Herein, we investigated whether CNP inhibits leukocyte recruitment and platelet aggregation and thereby exerts a potential antiinflammatory influence on the blood vessel wall. We assessed the effects of CNP on leukocyte-endothelial cell interactions in mouse mesenteric postcapillary venules in vivo in animals with high basal leukocyte activation (endothelial nitric oxide synthase knockout mice, eNOS-/-) or under acute inflammatory conditions (induced by interleukin-1 or histamine). CNP suppressed basal leukocyte rolling in eNOS-/- mice in a rapid, reversible, and concentration-dependent manner. These effects of CNP were mimicked by the selective natriuretic peptide receptor-C agonist cANF4-23. CNP also suppressed leukocyte rolling induced by IL-1 or histamine, inhibited platelet-leukocyte interactions, and prevented thrombin-induced platelet aggregation of human blood. Furthermore, analysis of human umbilical vein endothelial cells, leukocytes, and platelets revealed that CNP selectively attenuates expression of P-selectin. Thus, CNP is a modulator of acute inflammation in the blood vessel wall characterized by leukocyte and platelet activation. These antiinflammatory effects appear to be mediated, at least in part, via suppression of P-selectin expression. These observations suggest that endothelial CNP might maintain an anti-atherogenic influence on the blood vessel wall and represent a target for therapeutic intervention in inflammatory cardiovascular disorders. endothelium | natriuretic peptide receptor type C | atherosclerosis | thrombosis

  3. Thermal injury induces impaired function in polymorphonuclear neutrophil granulocytes and reduced control of burn wound infection

    PubMed Central

    Calum, H; Moser, C; Jensen, P Ř; Christophersen, L; Maling, D S; van Gennip, M; Bjarnsholt, T; Hougen, H P; Givskov, M; Jacobsen, G K; Hřiby, N

    2009-01-01

    Severe thermal injury induces immunosuppression, involving all parts of the immune system, especially when large fractions of the total body surface area are affected. An animal model was established to characterize the burn-induced immunosuppression. In our novel mouse model a 6% third-degree burn injury was induced in mice with a hot-air blower. The third-degree burn was confirmed histologically. The mice were allocated into five groups: control, shave, burn, infection and burn infection group. At 48 h, a decline in the concentration of peripheral blood leucocytes was observed in the group of mice with burn wound. The reduction was ascribed to the decline in concentration of polymorphonuclear neutrophil leucocytes and monocytes. When infecting the skin with Pseudomonas aeruginosa, a dissemination of bacteria was observed only in the burn wound group. Histological characterization of the skin showed a more polymorphonuclear neutrophil granulocytes (PMNs)-dominated inflammation in the group of mice with infected burn wound compared with the with burn wound group. In contrast, a higher degree of inflammation was observed in the burn wound group compared with the group of mice with infected burn wound. Furthermore, the oxidative burst and the phagocytic capacity of the PMNs were reduced in the group of mice with burn wound. Using this novel mouse model of thermal injury a decline of peripheral leucocytes was observed, whereas the increased local inflammatory response at the site of infection showed reduced capacity to contain and eliminate the infection. PMID:19210518

  4. The Multiple Chemokine–Binding Bovine Herpesvirus 1 Glycoprotein G (BHV1gG) Inhibits Polymorphonuclear Cell but Not Monocyte Migration into Inflammatory Sites

    PubMed Central

    Liu, Zheng; Bethunaickan, Ramalingam; Sahu, Ranjit; Brenner, Max; Laragione, Teresina; Gulko, Percio S; Davidson, Anne

    2013-01-01

    Chemokines facilitate the recruitment of inflammatory cells into tissues, contributing to target organ injury in a wide range of inflammatory and autoimmune diseases. Targeting either single chemokines or chemokine receptors alters the progression of disease in animal models of rheumatoid arthritis and lupus with varying degrees of efficacy, but clinical trials in humans have been less successful. Given the redundancy of chemokine–chemokine receptor interactions, targeting of more than one chemokine may be required to inhibit active inflammatory disease. To test the effects of multiple chemokine blockade in inflammation, we generated an adenovirus expressing bovine herpesvirus 1 glycoprotein G (BHV1gG), a viral chemokine antagonist that binds to a wide spectrum of murine and human chemokines, fused to the fragment crystallizable (Fc) portion of murine immunoglobulin (IgG)2a. Administration of the adenovirus significantly inhibited thioglycollate-induced migration of polymorphonuclear leukocytes into the peritoneal cavity of BALB/c mice and reduced both clinical severity and articular damage in K/BxN serum transfer-induced arthritis. However, treatment with BHV1gG-Ig fusion protein did not prevent monocyte infiltration into the peritoneum in the thioglycollate model and did not prevent renal monocyte infiltration or nephritis in lupus-prone NZB/W mice. These observations suggest that the simultaneous inhibition of multiple chemokines by BHV1gG has the potential to interfere with acute inflammatory responses mediated by polymorphonuclear leukocytes, but is less effective in chronic inflammatory disease mediated by macrophages.

  5. Stimulation of Ca2+-dependent chemiluminescence in rat polymorphonuclear leucocytes by polystyrene beads and the non-lytic action of complement.

    PubMed Central

    Hallett, M B; Luzio, J P; Campbell, A K

    1981-01-01

    (1) Chemiluminenscence of rat polymorphonuclear leukocytes was stimulated by a phagocytic stimulus, latex beads (diameter = 1.01 micrometer). The maximum chemiluminescent intensity increased with bead concentration in the range 0.2--20 x 10(9) beads/ml. This response was abolished in the absence of extracellular Ca2+ (1 mM EGTA). (2) Chemiluminescence could also be stimulated by the Ca2+ ionophore A23187 in the presence of extra-cellular calcium. (3) Addition of human serum, as a source of complement, to rat polymorphonuclear leukocytes preincubated with anti-5'-nucleotidase serum resulted in a rapid stimulation of chemiluminescence, after a lag of about 40 s. (4) The stimulation of chemiluminescence by antibody plus complement was not the result of cell lysis because (i) no significant release of lactate dehydrogenase was detected at the time of the chemiluminescent response (ii) chemiluminescence was associated with the cells and not the surrounding media (iii) cell lysis did not produce chemiluminescence. (5) Chemiluminescence stimulated by antibody plus complement or by beads was inhibited by the 'calmodulin-blocker', trifluoperazine (50% inhibiton with approximately 20--30 microM). (6) Cu2+ (10(-4) M), which can inhibit C9 action, inhibited the rapid rise in chemiluminescence induced by antibody plus complement, but not the bead-induced chemiluminescence. (7) Depletion of C9 from human serum markedly inhibited the complement induced chemiluminescence response. Addition of purified C9 restored the response. (8) It was concluded that formation of the terminal complement attack complex at the surface of rat polymorphonuclear leucocytes induces a Ca2+-dependent chemiluminescence in the cells, in the absence of cell lysis. PMID:7319554

  6. The road less traveled: regulation of leukocyte migration across vascular and lymphatic endothelium by galectins.

    PubMed

    Thiemann, Sandra; Baum, Linda G

    2011-02-01

    Leukocyte entry from the blood into inflamed tissues, exit into the lymphatics, and migration to regional lymph nodes are all crucial processes for mounting an effective adaptive immune response. Leukocytes must cross two endothelial cell layers, the vascular and the lymphatic endothelial cell layers, during the journey from the blood to the lymph node. The proteins and cellular interactions which regulate leukocyte migration across the vascular endothelium are well studied; however, little is known about the factors that regulate leukocyte migration across the lymphatic endothelium. Here, we will summarize evidence for a role for galectins, a family of carbohydrate-binding proteins, in regulating leukocyte migration across the vascular endothelium and propose that galectins are also involved in leukocyte migration across the lymphatic endothelium. PMID:20859666

  7. Expression of selected proteins of the extrinsic and intrinsic pathways of apoptosis in human leukocytes exposed to N-nitrosodimethylamine.

    PubMed

    Iwaniuk, A; Jab?o?ska, E; Jab?o?ski, J; Ratajczak-Wrona, W; Garley, M

    2015-06-01

    N-nitrosodimethylamine (NDMA) is a xenobiotic widespread in human environment capable of regulating the lifespan of immune cells. In this study, we examined the roles of the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)/death receptor 5 (DR5) complex and the Fas molecule in the induction of the extrinsic apoptosis pathway in human neutrophils (polymorphonuclear neutrophils (PMNs)) and peripheral blood mononuclear cells (PBMCs) exposed to NDMA. Also we assessed these proteins ability to trigger the intrinsic apoptosis pathway in those cells. For this purpose, we examined the expression of Fas-associated protein with death domain, truncated Bid (tBid) proteins, and apoptogenic factors such as apoptosis-inducing factor, Smac/Diablo, Omi/HtrA2, and caspase-3 as an indication of accomplished apoptosis phenomenon. PMNs and PBMCs were isolated from whole blood by density gradient centrifugation using Polymorphrep. Apoptotic cells were assessed with flow cytometry using a ready-made kit. The expression of proapoptotic molecules was investigated by Western blot analysis of PMNs and PBMCs treated with NDMA and/or rhTRAIL. The obtained results confirm the proapoptotic effects of NDMA on the examined human leukocytes and indicate an active participation of the TRAIL/DR5 complex and Fas protein in the process of apoptosis. Moreover, the research revealed distinct mechanisms of intrinsic apoptosis pathway activation between PMNs and PBMCs exposed to NDMA, as confirmed by the different levels of tBid, Smac/Diablo, Omi/HtrA2, and caspase-3 expression in those cells. PMID:25304970

  8. Individual radiosensitivity and its daily variations. [leukocyte reaction to epinephrine load

    NASA Technical Reports Server (NTRS)

    Druzhinin, Y. N.; Grigoryev, Y. G.; Podluzhnaya, G. N.; Pospishil, M.

    1974-01-01

    The effectiveness of determining individual radiosensitivity of rats by total gas exchange measurements, studies of Na/K content in urine, and the reaction of leukocytes to intra-abdominal administration of epinephrine, was studied. The most indicative results of predicting individual reaction to radiation were obtained by the leukocyte reaction to epinephrine load; however, changes in the leukocyte content of peripheral blood after epinephrine administration depended on the initial level during the day.

  9. Adhesion molecule cascades direct lymphocyte recirculation and leukocyte migration during inflammation

    Microsoft Academic Search

    Douglas A. Steeber; Thomas F. Tedder

    2000-01-01

    Leukocyte inter actions with vascular endothelium are high lyorchestrated processes that include the capture of free-flow\\u000a ing leukocytes from the blood with subsequent leukocyte rolling, arrest, firm adhesion, and ensuing diapedesis. These interactions\\u000a occur under high shear stresses within venules and depend on multiple families of adhesion molecules. Many of the adhesion\\u000a molecules involved are now identified. In addition, precise

  10. Effect of cytochalasin B on intracellular free calcium concentration in human polymorphnuclear leukocytes after repeated stimulation with n -formyl-methionyl-leucyl-phenylalanine

    Microsoft Academic Search

    D. Nowak; P. Bialasiewicz; A. Antczak; M. Krol; G. Piasecka

    1995-01-01

    Summary  Cytochalasin B can influence various functions of human polymorphonuclear leukocytes, including chemotaxis, lysosomal enzyme\\u000a release, and reactive oxygen species generation. In this study we investigated the effect of cytochalasin B on the increase\\u000a in intracellular free calcium concentration after three consecutive additions of 10?7 MN-formyl-methionyl-leucyl-phenylalanine. The interval between stimulations was 5 min. Intracellular free calcium concentration\\u000a was monitored using the

  11. [XE-2100: platelet clumps or polymorphonuclear neutrophil aggregates, that is the question].

    PubMed

    Dubois-Galopin, Frédérique; Lamballais, Florence; Rapatel, Chantal; Berger, Marc Gabriel

    2010-01-01

    Leukoagglutination of polymorphonuclear neutrophils is described as a rare phenomenon but its incidence is certainly underestimated. We report here 4 cases showing why this phenomenon is sometimes difficult to detect. In each case, the only flag reported from the hematology analyser XE-2100 (Sysmex corporation) was 'Platelet clumps?'. The graphic patterns were not suggestive of leukoagglutination but rather evoked platelet clumps. The 4 white blood cell differential scattergrams were absolutely normal and the 4 cases were not associated with leukoneutropenia. Two patients had clumps located only at the edges of the smears, and automated imaging processes, scanning only the center of the smears, may also contribute to the failure of recognising this phenomenon. PMID:20650744

  12. Decreased polymorphonuclear leucocyte chemotactic response to leukotriene B4 in cystic fibrosis.

    PubMed Central

    Lawrence, R H; Sorrelli, T C

    1992-01-01

    Evidence that leukotriene B4 (LTB4) is a significant inflammatory mediator in chronic pseudomonal respiratory disease was sought in adolescents and young adults with cystic fibrosis. Specific chemotaxis of peripheral blood polymorphonuclear leucocytes (PMN) was used as an indirect measure of remote in vivo exposure to LTB4. PMN from 17 patients showed a significant decrease in chemotaxis to 10(-7)-10(-9) M LTB4, but normal responses to 10(-8) M n-formyl-methionyl-leucyl-phenylalanine and 4 mg/ml casein, when compared with 17 healthy age- and sex-matched controls. This result is consistent with chronic production of LTB4, and specific deactivation of circulating PMN receptors for LTB4 in patients with cystic fibrosis. Pharmacologic inhibition of LTB4 production in vivo may help elucidate its role in the pathogenesis of lung damage in cystic fibrosis. PMID:1322257

  13. Quantitative analysis of human herpesvirus-6 and human cytomegalovirus in blood and saliva from patients with acute leukemia.

    PubMed

    Nefzi, Faten; Ben Salem, Nabil Abid; Khelif, Abderrahim; Feki, Salma; Aouni, Mahjoub; Gautheret-Dejean, Agnčs

    2015-03-01

    Human herpesvirus-6 (HHV-6) and human cytomegalovirus (HCMV) DNAs were quantified by real-time PCR assays in blood and saliva obtained from 50 patients with acute leukemia at the time of diagnosis (50 of each matrix), aplasia (65 of each matrix), remission (55 of each matrix), and relapse (20 of each matrix) to evaluate which biological matrix was more suitable to identify a viral reactivation, search for a possible link between HHV-6 and HCMV reactivations, and evaluate the relations between viral loads and count of different leukocyte types in blood. The median HHV-6 loads were 136; 219; 226, and 75 copies/million cells in blood at diagnosis, aplasia, remission and relapse, respectively. The HCMV loads were 193 and 317 copies/million cells in blood at diagnosis and remission. In the saliva samples, the HHV-6 loads were 22,165; 15,238; 30,214, and 17,454 copies/million cells at diagnosis, aplasia, remission, and relapse, respectively. The HCMV loads were 8,991; 1,461; 2,980, and 4,283 copies/million cells at diagnosis, aplasia, remission, and relapse, respectively. The HHV-6 load in the blood was correlated to the counts of polymorphonuclear leukocytes (R(2) ?=?0.5; P?blood in the detection of HHV-6 or HCMV reactivations. The HHV-6 and HCMV reactivations were linked only in saliva. PMID:25163462

  14. Carp thrombocyte phagocytosis requires activation factors secreted from other leukocytes.

    PubMed

    Nagasawa, Takahiro; Somamoto, Tomonori; Nakao, Miki

    2015-10-01

    Thrombocytes are nucleated blood cells in non-mammalian vertebrates, which were recently focused on not only as hemostatic cells but also as immune cells with potent phagocytic activities. We have analyzed the phagocytic activation mechanisms in common carp (Cyprinus carpio) thrombocytes. MACS-sorted mAb(+) thrombocytes showed no phagocytic activity even in the presence of several stimulants. However, remixing these thrombocytes with other anti-thrombocyte mAb(-) leukocyte populations restored their phagocytic activities, indicating that carp thrombocyte phagocytosis requires an appropriate exogenous stimulation. Culture supernatant from anti-thrombocyte mAb(-) leukocytes harvested after PMA or LPS stimulation, but not culture supernatant from unstimulated leukocytes, could activate thrombocyte phagocytosis. This proposed mechanism of thrombocyte phagocytosis activation involving soluble factors produced by activated leukocytes suggests that thrombocyte activation is restricted to areas proximal to injured tissues, ensuring suppression of excessive thrombocyte activation and a balance between inflammation and tissue repair. PMID:25978929

  15. Human uterine leukocytes and pregnancy.

    PubMed

    Trundley, A; Moffett, A

    2004-01-01

    In human pregnancy, the embryo implants into the specialized mucosal wall of the uterus (decidua) and the placenta starts to form. Cells from the placenta (trophoblasts) invade into the uterine mucosa in order to open up maternal uterine arteries to ensure an adequate supply of blood to the developing fetus. The trophoblasts have a unique immunological phenotype compared to most cells especially with regard to their expression of major histocompatibility complex (MHC) antigens. On the other side of the interaction, the uterine mucosa (endometrium) differentiates in preparation for implantation. One of the changes that takes place is the appearance in the endometrium of a large number of maternal leukocytes in the final part of the menstrual cycle. If pregnancy ensues, these leukocytes continue to increase in number and are found in close contact with trophoblasts. The composition of this population of maternal immune cells is unusual compared to that seen at other mucosal sites. A lot of research has focused on whether maternal T-cell responses are suppressed or modified during pregnancy. Research has also concentrated on the specialized uterine natural killer (NK) cells, which are found in the decidua in large numbers during early pregnancy. These uterine NK cells have been shown to express receptors for trophoblast MHC antigens, but their role in pregnancy is still mysterious. The purpose of this review is to give an overview of what is known about the immunology at the implantation site and also to provide an update of some of the most recent findings in this field. PMID:14651517

  16. Expression of the human NRAMP1 gene in professional primary phagocytes: studies in blood cells and in HL-60 promyelocytic leukemia.

    PubMed

    Cellier, M; Shustik, C; Dalton, W; Rich, E; Hu, J; Malo, D; Schurr, E; Gros, P

    1997-01-01

    In the mouse, mutations at the natural resistance-associated macrophage protein 1 (Nramp1) gene abrogate resistance to infection with antigenically unrelated intracellular parasites such as Mycobacterium, Salmonella, and Leishmania. Nramp1 expression is restricted to reticuloendothelial organs and peripheral blood leukocytes, where the protein may function as a membrane transporter of an as yet to be identified substrate. To identify the human blood cell type(s) expressing NRAMP1 mRNA and determine how Nramp1 expression is regulated in these cells, we have examined separated populations of peripheral blood leukocytes and in vitro cell lines. We observed that polymorphonuclear leukocytes (PMN) are the major site of NRAMP1 expression, followed to a lesser degree by monocytes (MN). Migration of MN to tissues (alveolar macrophages) or maturation in vitro (long-term culture) was associated with a higher level of NRAMP1 expression compared with blood MN. Northern analyses of RNA from model cultured cells showed absence of NRAMP1 expression in transformed cell lines from either erythroid or lymphoid T or B lineages as well as progenitors of the monocyte/macrophage pathway (KG1, U937, THP1), and the HL-60 promyelocytic leukemia. Induction of differentiation of HL-60 cells toward either the monocyte/macrophage (vitamin D3, phorbol ester) or the granulocyte pathways (DMF, DMSO), as measured by induction of IL8-Rb, c-FMS, and CD14 marker gene expression, was concomitant with a strong induction of NRAMP1 expression. These results suggest that NRAMP1 expression is specific to the myeloid lineage and is acquired during the maturation of PMN and MN. The possibility that NRAMP1 may be a component of the phagosomal/endosomal apparatus common to PMN and MN is discussed. PMID:9000542

  17. Red blood cells serve as intravascular carriers of myeloperoxidase.

    PubMed

    Adam, Matti; Gajdova, Silvie; Kolarova, Hana; Kubala, Lukas; Lau, Denise; Geisler, Anne; Ravekes, Thorben; Rudolph, Volker; Tsao, Philip S; Blankenberg, Stefan; Baldus, Stephan; Klinke, Anna

    2014-09-01

    Myeloperoxidase (MPO) is a heme enzyme abundantly expressed in polymorphonuclear neutrophils. MPO is enzymatically capable of catalyzing the generation of reactive oxygen species (ROS) and the consumption of nitric oxide (NO). Thus MPO has both potent microbicidal and, upon binding to the vessel wall, pro-inflammatory properties. Interestingly, MPO - a highly cationic protein - has been shown to bind to both endothelial cells and leukocyte membranes. Given the anionic surface charge of red blood cells, we investigated binding of MPO to erythrocytes. Red blood cells (RBCs) derived from patients with elevated MPO plasma levels showed significantly higher amounts of MPO by flow cytometry and ELISA than healthy controls. Heparin-induced MPO-release from patient-derived RBCs was significantly increased compared to controls. Ex vivo experiments revealed dose and time dependency for MPO-RBC binding, and immunofluorescence staining as well as confocal microscopy localized MPO-RBC interaction to the erythrocyte plasma membrane. NO-consumption by RBC-membrane fragments (erythrocyte "ghosts") increased with incrementally greater concentrations of MPO during incubation, indicating preserved catalytic MPO activity. In vivo infusion of MPO-loaded RBCs into C57BL/6J mice increased local MPO tissue concentrations in liver, spleen, lung, and heart tissue as well as within the cardiac vasculature. Further, NO-dependent relaxation of aortic rings was altered by RBC bound-MPO and systemic vascular resistance significantly increased after infusion of MPO-loaded RBCs into mice. In summary, we find that MPO binds to RBC membranes in vitro and in vivo, is transported by RBCs to remote sites in mice, and affects endothelial function as well as systemic vascular resistance. RBCs may avidly bind circulating MPO, and act as carriers of this leukocyte-derived enzyme. PMID:24976018

  18. Transmigration of polymorphnuclear neutrophils and monocytes through the human blood-cerebrospinal fluid barrier after bacterial infection in vitro

    PubMed Central

    2013-01-01

    Background Bacterial invasion through the blood-cerebrospinal fluid barrier (BCSFB) during bacterial meningitis causes secretion of proinflammatory cytokines/chemokines followed by the recruitment of leukocytes into the CNS. In this study, we analyzed the cellular and molecular mechanisms of polymorphonuclear neutrophil (PMN) and monocyte transepithelial transmigration (TM) across the BCSFB after bacterial infection. Methods Using an inverted transwell filter system of human choroid plexus papilloma cells (HIBCPP), we studied leukocyte TM rates, the migration route by immunofluorescence, transmission electron microscopy and focused ion beam/scanning electron microscopy, the secretion of cytokines/chemokines by cytokine bead array and posttranslational modification of the signal regulatory protein (SIRP) ? via western blot. Results PMNs showed a significantly increased TM across HIBCPP after infection with wild-type Neisseria meningitidis (MC58). In contrast, a significantly decreased monocyte transmigration rate after bacterial infection of HIBCPP could be observed. Interestingly, in co-culture experiments with PMNs and monocytes, TM of monocytes was significantly enhanced. Analysis of paracellular permeability and transepithelial electrical resistance confirmed an intact barrier function during leukocyte TM. With the help of the different imaging techniques we could provide evidence for para- as well as for transcellular migrating leukocytes. Further analysis of secreted cytokines/chemokines showed a distinct pattern after stimulation and transmigration of PMNs and monocytes. Moreover, the transmembrane glycoprotein SIRP? was deglycosylated in monocytes, but not in PMNs, after bacterial infection. Conclusions Our findings demonstrate that PMNs and monoctyes differentially migrate in a human BCSFB model after bacterial infection. Cytokines and chemokines as well as transmembrane proteins such as SIRP? may be involved in this process. PMID:23448224

  19. Junctional Adhesion Molecule-C Regulates the Early Influx of Leukocytes into Tissues during Inflammation1

    Microsoft Academic Search

    Michel Aurrand-Lions; Chrystelle Lamagna; John P. Dangerfield; Shijun Wang; Pedro Herrera; Sussan Nourshargh; Beat A. Imhof

    2005-01-01

    Leukocyte recruitment from blood to inflammatory sites occurs in a multistep process that involves discrete molecular interactions between circulating and endothelial cells. Junctional adhesion molecule (JAM)-C is expressed at different levels on endothelial cells of lymphoid organs and peripheral tissues and has been proposed to regulate neutrophil migration by its interaction with the leukocyte integrin Mac-1. In the present study,

  20. Effects of Eugenol on Polymorphonuclear Cell Migration and Chemiluminescence

    Microsoft Academic Search

    P. G. Fotos; C. J. Woolverton; K. Van Dyke; R. L. Powell

    1987-01-01

    In this study, the effects of eugenol on human polymorphonuclear (PMN) cell migration and chemiluminescence were examined in vitro. Utilizing zymosan-activated serum or crude Bacteroides sonicate fractions as chemotractants, we found that eugenol inhibits PMN migration at 6.6 x 10-2to 6.6 x 10-5mol\\/L (P<0.05). Also, similar effects were observed in PMNs pre-incubated in eugenol. Regardless of concentration, eugenol was not

  1. Fibrinogen modulates leukocyte recruitment in vivo during the acute inflammatory response.

    PubMed

    Vitorino de Almeida, V; Silva-Herdade, A; Calado, A; Rosário, H S; Saldanha, C

    2015-01-01

    Besides playing an important role in blood hemostases, fibrinogen also regulates leukocyte function in inflammation. Our previous in vitro studies showed that the adhesive behaviour of the neutrophil is modulated by soluble fibrinogen when present at a physiological concentration. This led us to propose that this plasma glycoprotein might further influence leukocyte recruitment in vivo and thus contribute to the inflammatory response. To address this in vivo, leukocyte recruitment was here investigated under acute inflammatory conditions in the absence of soluble fibrinogen in the blood circulation. For such, intravital microscopy on mesentery post-capillary venules was performed on homozygous fibrinogen ? chain-deficient mice ((?-/-) mice). Acute inflammatory states were induced by perfusing platelet activating factor (PAF) over the exposed tissue. As control animals, two groups of mice expressing soluble fibrinogen in circulation were used, namely, C57BL/6 wild type animals and heterozygous fibrinogen ? chain-deficient mice ((?+/-) mice). Under acute inflammatory conditions, an abnormal pattern of recruitment was observed for leukocytes in homozygous (?-/-) mice in comparison to both control groups. In fact, the former exhibited a significantly decreased number of rolling leukocytes that nevertheless, migrated with increased rolling velocities when compared to leukocytes from control animals. Consistently, homozygous mice further displayed a diminished number of adherent leukocytes than the other groups. Altogether our observations led us to conclude that leukocyte recruitment in homozygous (?-/-) mice is compromised what strongly suggests a role for soluble fibrinogen in leukocyte recruitment in inflammation. PMID:23271199

  2. Dipeptidyl peptidase II and leukocyte cell death.

    PubMed

    Maes, Marie-Berthe; Martinet, Wim; Schrijvers, Dorien M; Van der Veken, Pieter; De Meyer, Guido R Y; Augustyns, Koen; Lambeir, Anne-Marie; Scharpé, Simon; De Meester, Ingrid

    2006-06-28

    Dipeptidyl peptidase (DPP) II (E.C. 3.4.14.2) is an intracellular protease that releases, preferably at acidic pH, N-terminal dipeptides from oligopeptides with Pro or Ala in the penultimate position. The natural substrates and the physiological role of DPPII remain unclear. The aim of the present study was to investigate the involvement of DPPII activity in different forms of cell death (apoptosis, necrosis and autophagy) in human leukocytes. We determined specific DPP activities in leukocytes. Compared to other subpopulations of peripheral blood mononuclear cells (PBMC), we observed relatively high DPPII specific activity in monocytic cells, opening new perspectives for further investigation of the DPPII functions. A second intriguing finding was that DPPII specific activity increased during necrosis, whereas induction of apoptosis or autophagy did not affect any of the dipeptidyl peptidase activities. Finally, we showed that inhibition of DPPII (>90%) using the in vitro applicable, highly potent (K(i) of 0.082+/-0.048 nM) and selective DPPII inhibitor UAMC00039, did not induce any form of cell death in leukocytes. These data are of importance for a more precise interpretation of the in vitro and in vivo effects of other dipeptidyl peptidase inhibitors. PMID:16725115

  3. Leukocyte filtration in lung transplantation.

    PubMed

    Kurusz, Mark; Roach, John D; Vertrees, Roger A; Girouard, Mark K; Lick, Scott D

    2002-05-01

    Controlled reperfusion of the transplanted lung has been used in nine consecutive patients to decrease manifestations of lung reperfusion injury. An extracorporeal circuit containing a roller pump, heat exchanger and leukodepleting filter is primed with substrate-enhanced reperfusion solution mixed with approximately 2000 ml of the patient's blood. This solution is slowly recirculated to remove leukocytes prior to reperfusion. When the pulmonary anastomoses are completed, the pulmonary artery is cannulated through the untied anastomosis using a catheter containing a pressure lumen for measurement of infusion pressure. An atrial clamp is left in place on the patient's native atrial cuff to decrease the risk of systemic air embolism during the brief period of reperfusion from the extracorporeal reservoir. During reperfusion, the water bath to the heat exchanger is kept at 35 degrees C and the flow rate for reperfusion solution is between 150 and 200 m/min, keeping the pulmonary artery pressure <14 mmHg. Eight of nine patients were ventilated on 40% inspired oxygen within a few hours of operation and 7/9 were extubated on or before postoperative day 1. Six of nine patients are long-term survivors. PMID:12009087

  4. Leukocytes in chemotactic-fragment-induced lung inflammation. Vascular emigration and alveolar surface migration.

    PubMed

    Shaw, J O

    1980-11-01

    Lung inflammation was induced in rabbits by intratracheal injections of chemotactic fragments obtained from zymosan-activated serum (CF-ZAS), and the route of vascular emigration and alveolar surface interaction of polymorphonuclear leukocytes (PMNs) and monocytes migrating into the lung was characterized by transmission (TEM) and scanning (SEM) electron-microscopic examination. Leukocytes migrated from capillaries and venules into the alveolar wall interstitium by adherence to the vascular endothelium and migration through the endothelial intracellular junction to attain a position between a reapposed endothelial cell junction and the vascular basement membrane. The cells then migrated into the interstitium through a narrow opening in the basement membrane. Leukocyte entrance into the alveolar space from the interstitium appeared to occur through small openings in the epithelial basement membrane at or near the Type I epithelial intercellular junction. Once in the alveolus, PMNs and macrophages demonstrated surface adherence and spreading along with evidence of migration, pseudopod extension, interalveolar pore transit, and retraction fiber formation. This study indicates the leukocyte influx into the alveolus in acute chemotactic-factor-induced inflammation is via a continuum of migrational activity, beginning at the pulmonary capillary endothelial surface and persisting on the alveolar epithelial surface. PMID:7435538

  5. Leukocytes in chemotactic-fragment-induced lung inflammation. Vascular emigration and alveolar surface migration.

    PubMed Central

    Shaw, J. O.

    1980-01-01

    Lung inflammation was induced in rabbits by intratracheal injections of chemotactic fragments obtained from zymosan-activated serum (CF-ZAS), and the route of vascular emigration and alveolar surface interaction of polymorphonuclear leukocytes (PMNs) and monocytes migrating into the lung was characterized by transmission (TEM) and scanning (SEM) electron-microscopic examination. Leukocytes migrated from capillaries and venules into the alveolar wall interstitium by adherence to the vascular endothelium and migration through the endothelial intracellular junction to attain a position between a reapposed endothelial cell junction and the vascular basement membrane. The cells then migrated into the interstitium through a narrow opening in the basement membrane. Leukocyte entrance into the alveolar space from the interstitium appeared to occur through small openings in the epithelial basement membrane at or near the Type I epithelial intercellular junction. Once in the alveolus, PMNs and macrophages demonstrated surface adherence and spreading along with evidence of migration, pseudopod extension, interalveolar pore transit, and retraction fiber formation. This study indicates the leukocyte influx into the alveolus in acute chemotactic-factor-induced inflammation is via a continuum of migrational activity, beginning at the pulmonary capillary endothelial surface and persisting on the alveolar epithelial surface. Images Figure 10 Figure 11 Figure 12 Figure 1 Figure 2 Figure 3 Figure 13 Figure 14 Figure 4 Figure 5 Figure 6 Figure 15 Figure 7 Figure 8 Figure 16 Figure 9 PMID:7435538

  6. The Road Less Traveled: Regulation of Leukocyte Migration Across Vascular and Lymphatic Endothelium by Galectins

    Microsoft Academic Search

    Sandra Thiemann; Linda G. Baum

    2011-01-01

    Leukocyte entry from the blood into inflamed tissues, exit into the lymphatics, and migration to regional lymph nodes are\\u000a all crucial processes for mounting an effective adaptive immune response. Leukocytes must cross two endothelial cell layers,\\u000a the vascular and the lymphatic endothelial cell layers, during the journey from the blood to the lymph node. The proteins\\u000a and cellular interactions which

  7. Quantitative reconstruction of leukocyte subsets using DNA methylation

    PubMed Central

    2014-01-01

    Background Cell lineage-specific DNA methylation patterns distinguish normal human leukocyte subsets and can be used to detect and quantify these subsets in peripheral blood. We have developed an approach that uses DNA methylation to simultaneously quantify multiple leukocyte subsets, enabling investigation of immune modulations in virtually any blood sample including archived samples previously precluded from such analysis. Here we assess the performance characteristics and validity of this approach. Results Using Illumina Infinium HumanMethylation27 and VeraCode GoldenGate Methylation Assay microarrays, we measure DNA methylation in leukocyte subsets purified from human whole blood and identify cell lineage-specific DNA methylation signatures that distinguish human T cells, B cells, NK cells, monocytes, eosinophils, basophils and neutrophils. We employ a bioinformatics-based approach to quantify these cell types in complex mixtures, including whole blood, using DNA methylation at as few as 20 CpG loci. A reconstruction experiment confirms that the approach could accurately measure the composition of mixtures of human blood leukocyte subsets. Applying the DNA methylation-based approach to quantify the cellular components of human whole blood, we verify its accuracy by direct comparison to gold standard immune quantification methods that utilize physical, optical and proteomic characteristics of the cells. We also demonstrate that the approach is not affected by storage of blood samples, even under conditions prohibiting the use of gold standard methods. Conclusions Cell mixture distributions within peripheral blood can be assessed accurately and reliably using DNA methylation. Thus, precise immune cell differential estimates can be reconstructed using only DNA rather than whole cells. PMID:24598480

  8. Hydrodynamic forces on a wall-bound leukocyte in small vessels due to red cells

    NASA Astrophysics Data System (ADS)

    Isfahani, Amir H. G.; Freund, Jonathan B.

    2010-11-01

    As part of the inflammation response, white blood cells (leukocytes) bind to the vessel wall before they transmigrate across the endothelium. The interactions between the wall-adhered leukocyte and flowing red blood cells (erythrocytes) play a critical role in this process. We provide a quantitative investigation of the forces exerted on a wall-bound leukocyte using a simulation tool that is based on a fast O(N N) boundary integral formulation. This permits simulations of red cells that are both realistically flexible and can approach to very close separation distances. The elastic membranes deform substantially but strongly resist surface dilatation. The no-slip condition is enforced both on the leukocyte and the round vessel walls. Vessel diameters from 10 to 20 microns are studied. At these scales the cellular-particulate nature of blood significantly affects the magnitude of the forces that the leukocyte experiences. For a tube hematocrit (cell volume fraction) of 25% and a spherical protrusion with a diameter 0.75 times that of the tube, the average forces are increased by about 40% and the local forces by more than 100% relative to those expected for a blood model homogenized by its effective viscosity. For a constant pressure gradient, the wall-bound leukocyte causes a blockage in the vessel. Different contact angles for the leukocyte as well as different mechanical properties for the erythrocytes are examined.

  9. Detection of deep venous thrombosis by indium-111 leukocyte scintigraphy

    SciTech Connect

    D'Alonzo, W.A. Jr.; Alavi, A.

    1986-05-01

    Indium-111-labeled leukocyte ((/sup 111/In)WBC) scintigraphy has been used successfully for detection of inflammation. Occasionally, noninflammatory collections of white blood cells such as hematomas or hemorrhage have been localized. We report a case in which unsuspected femoral deep venous thrombosis was diagnosed on an (/sup 111/In)WBC leukocyte scan performed for detection of osteomyelitis. Readers are advised to avoid interpreting all vascular (/sup 111/In)WBC localization as necessarily infectious. This may be of particular significance in patients with vascular grafts.

  10. Observations on phagocytosis of urate crystals by polymorphonuclear leucocytes.

    PubMed Central

    Rajan, K T

    1975-01-01

    The sequence of events that may initiate the inflammatory reaction in acute gout has been investigated with specific reference to phagocytosis of urate crystals by polymorphonuclear leucocytes and the results have shown (1) that neutrophil leucocytes avidly ingest microcrystals of sodium monourate, (2) that this causes the rapid degranulation and disintegration of the leucocytes, (3) that fresh leucocytes ingest the debris and crystals liberated by the dead cells, and in their turn degranulate and die, thus possibly establishing a vicious circle in the system. Images PMID:47739

  11. Observations on phagocytosis of urate crystals by polymorphonuclear leucocytes.

    PubMed

    Rajan, K T

    1975-02-01

    The sequence of events that may initiate the inflammatory reaction in acute gout has been investigated with specific reference to phagocytosis of urate crystals by polymorphonuclear leucocytes and the results have shown (1) that neutrophil leucocytes avidly ingest microcrystals of sodium monourate, (2) that this causes the rapid degranulation and disintegration of the leucocytes, (3) that fresh leucocytes ingest the debris and crystals liberated by the dead cells, and in their turn degranulate and die, thus possibly establishing a vicious circle in the system. PMID:47739

  12. Polymorphonuclear leucocyte motility in men with ankylosing spondylitis.

    PubMed Central

    Pease, C T; Fennell, M; Brewerton, D A

    1989-01-01

    The polymorphonuclear leucocyte (PMN) response to a chemotactic or chemokinetic stimulus is enhanced in men with ankylosing spondylitis (AS). This effect does not parallel the severity of disease activity or the size of the acute phase response, and it is independent of non-steroidal anti-inflammatory drug treatment. Polymorph function is normal in HLA-B27 positive brothers of probands with AS and in other HLA-B27 positive individuals in the absence of disease. Polymorph motility is also normal in patients with psoriasis vulgaris or Crohn's disease, indicating that enhanced PMN motility is not a non-specific consequence of all inflammatory disorders. PMID:2784306

  13. Hydrodynamic forces on a wall-bound leukocyte due to interactions with flowing red cells

    NASA Astrophysics Data System (ADS)

    Isfahani, Amir H. G.; Freund, Jonathan B.

    2011-11-01

    As part of both healthy and pathologically physiological mechanisms sphere-like white blood cells (leukocytes) adhere to the walls of small blood vessels. We use quantitative numerical simulations to compare the forces from flowing red blood cells on a wall-adhered leukocyte to a homogenized model of blood at the same flow conditions. We model the highly flexible red blood cells using a fast O (N log N) boundary integral formulation. These elastic membranes deform substantially but strongly resist surface dilatation. They enclose a higher than plasma viscosity hemoglobin solution. The no-slip condition is enforced on the stationary leukocyte as well as the vessel walls. Vessel diameters of 10 to 20 microns are studied. Different hematocrits, leukocyte shapes, and flow conditions are examined. In vessels comparable to the size of the cells, we show that the particulate character of blood significantly affects the magnitude of the forces that the leukocyte experiences, transiently increasing it well above the homogenized-blood prediction: for example, for a tube hematocrit of 25 % and a spherical protrusion with a diameter 0.75 that of the tube, the average forces are increased by about 40 % and the local forces by more than 100 % relative to those expected for a blood model homogenized by its effective viscosity.

  14. Protein-bound uremic toxins stimulate crosstalk between leukocytes and vessel wall.

    PubMed

    Pletinck, Anneleen; Glorieux, Griet; Schepers, Eva; Cohen, Gerald; Gondouin, Bertrand; Van Landschoot, Maria; Eloot, Sunny; Rops, Angelique; Van de Voorde, Johan; De Vriese, An; van der Vlag, Johan; Brunet, Philippe; Van Biesen, Wim; Vanholder, Raymond

    2013-12-01

    Leukocyte activation and endothelial damage both contribute to cardiovascular disease, a major cause of morbidity and mortality in CKD. Experimental in vitro data link several protein-bound uremic retention solutes to the modulation of inflammatory stimuli, including endothelium and leukocyte responses and cardiovascular damage, corroborating observational in vivo data. However, the impact of these uremic toxins on the crosstalk between endothelium and leukocytes has not been assessed. This study evaluated the effects of acute and continuous exposure to uremic levels of indoxylsulfate (IS), p-cresylsulfate (pCS), and p-cresylglucuronide (pCG) on the recruitment of circulating leukocytes in the rat peritoneal vascular bed using intravital microscopy. Superfusion with IS induced strong leukocyte adhesion, enhanced extravasation, and interrupted blood flow, whereas pCS caused a rapid increase in leukocyte rolling. Superfusion with pCS and pCG combined caused impaired blood flow and vascular leakage but did not further enhance leukocyte rolling over pCS alone. Intravenous infusion with IS confirmed the superfusion results and caused shedding of heparan sulfate, pointing to disruption of the glycocalyx as the mechanism likely mediating IS-induced flow stagnation. These results provide the first clear in vivo evidence that IS, pCS, and pCG exert proinflammatory effects that contribute to vascular damage by stimulating crosstalk between leukocytes and vessels. PMID:24009240

  15. Antibacterial 15-kDa protein isoforms (p15s) are members of a novel family of leukocyte proteins.

    PubMed

    Levy, O; Weiss, J; Zarember, K; Ooi, C E; Elsbach, P

    1993-03-15

    We have previously described the isolation and initial characterization of functionally distinct 15-kDa protein isoforms (p15s) from rabbit polymorphonuclear leukocytes (PMN) that bind with high affinity to Escherichia coli and modulate the antibacterial actions of other leukocyte proteins on this Gram-negative bacterium. We now report the cloning and sequencing of two distinct cDNAs from a rabbit bone marrow library that encode p15s differing at only 2 residues (His-3, Arg-88 versus Arg-3, Trp-88). Tryptophan-directed chemical cleavage of two isoforms purified from a single rabbit confirms the existence of multiple isoforms with distinct function and primary structure in a single rabbit. The p15 cDNAs encode putative signal sequences and studies of cellular and subcellular localization indicate that the p15s are granule-associated proteins of PMN. Both purified isoforms bind avidly to lipopolysaccharide (LPS), the major component of the Gram-negative bacterial outer membrane. Analysis of the deduced primary structures of the p15s reveals homology to three other leukocyte proteins: CAP-18, an 18-kDa LPS-binding protein from rabbit PMN, pro-indolicidin, a 16-kDa precursor of an antibacterial peptide of bovine PMN, and cathelin, an 11-kDa cysteine protease inhibitor from porcine leukocytes, suggesting the existence of a novel family of leukocyte proteins with LPS-binding, antimicrobial, and protease-inhibitory activities. PMID:8449963

  16. Flow resistance and drag forces due to multiple adherent leukocytes in postcapillary vessels.

    PubMed Central

    Chapman, G B; Cokelet, G R

    1998-01-01

    Computational fluid dynamics was used to model flow past multiple adherent leukocytes in postcapillary size vessels. A finite-element package was used to solve the Navier-Stokes equations for low Reynolds number flow of a Newtonian fluid past spheres adhering to the wall of a cylindrical vessel. We determined the effects of sphere number, relative geometry, and spacing on the flow resistance in the vessel and the fluid flow drag force acting to sweep the sphere off the vessel wall. The computations show that when adherent leukocytes are aligned on the same side of the vessel, the drag force on each of the interacting leukocytes is less than the drag force on an isolated adherent leukocyte and can decrease by up to 50%. The magnitude of the reduction depends on the ratio of leukocyte to blood vessel diameter and distance between adherent leukocytes. However, there is an increase in the drag force when leukocytes adhere to opposite sides of the vessel wall. The increase in resistance generated by adherent leukocytes in vessels of various sizes is calculated from the computational results. The resistance increases with decreasing vessel size and is most pronounced when leukocytes adhere to opposite sides of the vessel. PMID:9635783

  17. Scaling deterministic lateral displacement arrays for high throughput and dilution-free enrichment of leukocytes

    NASA Astrophysics Data System (ADS)

    Inglis, David W.; Lord, Megan; Nordon, Robert E.

    2011-05-01

    A disposable device for fractionation of blood into its components that is simple to operate and provides throughput of greater than 1 mL min-1 is highly sought after in medical diagnostics and therapies. This paper describes a device with parallel deterministic lateral displacement devices for enrichment of leukocytes from blood. We show capture of 98% and approximately ten-fold enrichment of leukocytes in whole blood. We demonstrate scaling up through the integration of six parallel devices to achieve a flow rate of 115 µL of undiluted blood per minute per atmosphere of applied pressure.

  18. Imaging leukocytes in vivo with third harmonic generation microscopy

    NASA Astrophysics Data System (ADS)

    Tsai, Cheng-Kun; Chen, Chien-Kuo; Chen, Yu-Shing; Wu, Pei-Chun; Hsieh, Tsung-Yuan; Liu, Han-Wen; Yeh, Chiou-Yueh; Lin, Win-Li; Chia, Jean-San; Liu, Tzu-Ming

    2013-02-01

    Without a labeling, we demonstrated that lipid granules in leukocytes have distinctive third harmonic generation (THG) contrast. Excited by a 1230nm femtosecond laser, THG signals were generated at a significantly higher level in neutrophils than other mononuclear cells, whereas signals in agranular lymphocytes were one order smaller. These characteristic THG features can also be observed in vivo to trace the newly recruited leukocytes following lipopolysaccharide (LPS) challenge. Furthermore, using video-rate THG microscopy, we also captured images of blood cells in human capillaries. Quite different from red-blood-cells, every now and then, round and granule rich blood cells with strong THG contrast appeared in circulation. The corresponding volume densities in blood, evaluated from their frequencies of appearance and the velocity of circulation, fall within the physiological range of human white blood cell counts. These results suggested that labeling-free THG imaging may provide timely tracing of leukocyte movement and hematology inspection without disturbing the normal cellular or physiological status.

  19. Association between diabetes complications and leukocyte counts in Iranian patients

    PubMed Central

    Moradi, Sedigheh; Kerman, Scott Reza Jafarian; Rohani, Farzaneh; Salari, Fereshteh

    2012-01-01

    Background The long term complications of diabetes can be fatal. They are also renowned for being an economic burden. Previous studies have demonstrated a relationship between inflammatory markers and complications of diabetes. The objective of this study was to evaluate the relationship between leukocyte counts and these complications. Methods The study included 184 patients diagnosed with type 2 diabetes. The study was carried out in Iran during 2007 and 2008. Data collected on the subjects were as follows: age, gender, weight, height, blood pressure, smoking history, lipid profile including low density lipoprotein (LDL), high density lipoprotein (HDL), total cholesterol, triglycerides, and leukocyte count, albuminuria, and retinopathy. Furthermore, information on cardiac history for 100 patients was collected. The subjects were split into two groups according to their leukocyte levels: low (?7000/mm3) and high (>7000/mm3); and then analyzed by Student’s t-test or Mann–Whitney U-test as appropriate. Results The average leukocyte count in these patients was 7594 ± 1965/mm3. Leukocyte count was significantly different in patients with and without retinopathy and albuminuria (P < 0.0001). According to this analysis, a leukocyte count of 6750/mm3 with a sensitivity of 80.2% and a specificity of 56.4%, and a count of 7550/mm3 with a sensitivity of 63.2% and a specificity of 74.6% indicated at least one diabetes complication. Conclusion An elevated leukocyte count even within the normal range was associated with chronic complications in type 2 diabetes. PMID:22334791

  20. Effect of permethrin insecticide on rat polymorphonuclear neutrophils.

    PubMed

    Gabbianelli, Rosita; Falcioni, Maria Letizia; Nasuti, Cinzia; Cantalamessa, Franco; Imada, Isuke; Inoue, Masayasu

    2009-12-10

    Polymorphonuclear neutrophils are professional phagocytes whose efficacy depends on a multicomponent NADPH oxidase for generating superoxide anions and bacterial killing. They can be primed and activated by different agents that can impair oxidative burst and phagocytosis with opposite effects: reduced capability to destroy bacteria or hyperactivation that induces the generation of large quantities of toxic reactive oxygen species, which can damage surrounding tissue and participate in inflammation. The present study was designed to evaluate the effect of sub-chronic (60 days) permethrin treatment (1/10 DL(50)) on rat polymorphonuclear neutrophils respiratory burst. The results show that permethrin treatment increases superoxide anion production (33 times) and the activity of hydrogen peroxide-myeloperoxidase system (67 times). In vitro experiments suggest that this effect can be related to permethrin priming and to physico-chemical changes at the plasma membrane level of neutrophils. The antioxidant supplementation with Vitamin E and coenzyme Q(10) can protect against the abnormal respiratory burst in rat treated with permethrin. The in vitro studies show that neutrophil apoptosis begins soon after 1h of incubation with permethrin (0.725% of total cells) or its metabolites (3-phenoxybenzyl alcohol, 3-phenoxybenzaldehyde and 3-phenoxybenzoic acid 1.36, 2.26 and 1.3 of total cells, respectively) and that the level of apoptotic cells is very low. In conclusion, immunotoxicity of permethrin measured in rats could prompt future studies on the consequences of chronic insecticide exposure. PMID:19772857

  1. Initial blood storage experiment

    NASA Technical Reports Server (NTRS)

    Surgenor, Douglas MACN.

    1988-01-01

    The possibility of conducting experiments with the formed elements of the blood under conditions of microgravity opens up important opportunities to improve the understanding of basic formed element physiology, as well as, contribution to improved preservation of the formed elements for use in transfusion. The physiological, biochemical, and physical changes of the membrane of the erythrocyte, platelet, and leukocyte was studied during storage under two specific conditions: standard blood bank conditions and microgravity, utilizing three FDA approved plastic bags. Storage lesions; red cell storage on Earth; platelet storage on Earth; and leukocyte storage Earth were examined. The interaction of biomaterials and blood cells was studied during storage.

  2. Extravasation of leukocytes in comparison to tumor cells

    PubMed Central

    Strell, Carina; Entschladen, Frank

    2008-01-01

    The multi-step process of the emigration of cells from the blood stream through the vascular endothelium into the tissue has been termed extravasation. The extravasation of leukocytes is fairly well characterized down to the molecular level, and has been reviewed in several aspects. Comparatively little is known about the extravasation of tumor cells, which is part of the hematogenic metastasis formation. Although the steps of the process are basically the same in leukocytes and tumor cells, i.e. rolling, adhesion, transmigration (diapedesis), the molecules that are involved are different. A further important difference is that leukocyte interaction with the endothelium changes the endothelial integrity only temporarily, whereas tumor cell interaction leads to an irreversible damage of the endothelial architecture. Moreover, tumor cells utilize leukocytes for their extravasation as linkers to the endothelium. Thus, metastasis formation is indirectly susceptible to localization signals that are literally specific for the immune system. We herein compare the extravasation of leukocytes and tumor cells with regard to the involved receptors and the localization signals that direct the cells to certain organs and sites of the body. PMID:19055814

  3. Endothelial signaling in leukocyte transmigration

    Microsoft Academic Search

    Peter L. Hordijk

    2003-01-01

    Leukocyte transendothelial migration is a multistep process coordinated by chemokine receptors, integrins and cell adhesion\\u000a molecules. The interaction between leukocytes and endothelial cells is accompanied by bidirectional signaling in both cell\\u000a types, which is initiated following formation of specialized, “docking” structures. In recent years, it has become clear that\\u000a signaling in the endothelial cells importantly contributes to the transmigration process.

  4. Leukocyte activation, erythrocyte damage, lipid profile and oxidative stress imposed by high competition physical exercise in adolescents

    Microsoft Academic Search

    Alice Santos-Silva; Maria Irene Rebelo; Elisabeth Molnar Bayer Castro; Luis Belo; António Guerra; Carla Rego; Alexandre Quintanilha

    2001-01-01

    Background: The aim of this study was to evaluate and to compare the lipid profile and the levels of leukocyte activation, red blood cell (RBC) damage and of oxidative stress in two groups of adolescents, with similar body mass index: high competition swimmers and adolescents practising moderate regular physical exercise. Methods: As markers of leukocyte activation, we measured plasma lactoferrin,

  5. Leukocyte recruitment to peritoneal cavity of rats following formalin injection: role of tachykinin receptors.

    PubMed

    Santos, Júlia M M; Tatsuo, Maria Aparecida K F; Turchetti-Maia, Regina Maria M; Lisboa, Marcela C G; de Francischi, Janetti N

    2004-04-01

    The aim of this work was to verify whether formalin would induce leukocyte recruitment following intraperitoneal (i.p.) injection in rats. Formalin (1.25 - 2.5%) induced cell recruitment, which was concentration- and time-dependent (0 - 24 h). Two peaks of leukocyte recruitment were observed. The first peak (from 2 to 4 h) was characterized by a mixed polymorphonuclear and lymphocyte cell population (representing an increase of 100 - 220% and 55 - 60%, respectively), whereas the second peak was characterized by a marked increase in lymphocytes at 24 h (representing an increase of 230%). Pretreatment of animals with specific antagonists for neurokinin NK(1), NK(2), and NK(3) receptors (SR140333, SR48968, and SR142801 compounds, respectively) reduced the early leukocyte increase (representing a significant reduction of 65%, 51%, and 46%, respectively), whereas only the treatment with NK(2)-specific antagonist reduced the late cell increase induced by formalin injection (amounting to a significant reduction of 48%). These results suggested that substance P, neurokinin A, and neurokinin B release accounted for formalin-induced cell migratory activity. The anti-inflammatory drug dexamethasone also reduced cell recruitment, which was mainly related to a reduction in 79% of the neutrophils at 4 h following 1.25% formalin injection, suggesting also a release of lipid mediators (eicosanoids and/or platelet-activating factor) and/or cytokines/chemokines by the formalin injection. PMID:15107578

  6. [Peripheral leukocyte adhesion molecules in patients of Behçet's disease associated with active ocular lesions].

    PubMed

    Kaku, H; Mizukawa, H; Kishi, I; Yanagida, T; Inaba, G

    1994-06-01

    The sequential cascades in leukocyte adhesion and migration are important events in the development of inflammatory and immune responses in Behçet's disease. In an attempt to clarify the relation of active ocular lesions to inflammatory reactions covering entire body, we have detected LECAM-1, Mac-1 and CD44 expressed on the peripheral leukocytes in 24 Behçet patients and 15 healthy adults by flow cytometry. LECAM-1 and CD44 expression was dramatically decreased from the polymorphonuclear leukocytes (PMN) upon ocular attacks, whereas the changes in Mac-1 appeared not so striking. This tendency had continued into the recovery stage of ocular inflammation. T lymphocytes showed, on the other hand, no considerable variability in regard to the expression of cell surface LECAM-1, CD44 and also of Mac-1 even at the peak of active ocular inflammation. The results suggest that interaction between PMN and primed vascular endothelial cells might be apt to precede functional modifications of T lymphocytes, and it is also supposed that facilitation of activated PMN recruitment plays an essential role on ocular inflammation in Behçet's disease. PMID:7519790

  7. Is Chronic Asthma Associated with Shorter Leukocyte Telomere Length at Midlife?

    PubMed Central

    Shalev, Idan; Sears, Malcolm R.; Hancox, Robert J.; Lee Harrington, Hona; Houts, Renate; Moffitt, Terrie E.; Sugden, Karen; Williams, Benjamin; Poulton, Richie; Caspi, Avshalom

    2014-01-01

    Rationale: Asthma is prospectively associated with age-related chronic diseases and mortality, suggesting the hypothesis that asthma may relate to a general, multisystem phenotype of accelerated aging. Objectives: To test whether chronic asthma is associated with a proposed biomarker of accelerated aging, leukocyte telomere length. Methods: Asthma was ascertained prospectively in the Dunedin Multidisciplinary Health and Development Study cohort (n = 1,037) at nine in-person assessments spanning ages 9–38 years. Leukocyte telomere length was measured at ages 26 and 38 years. Asthma was classified as life-course-persistent, childhood-onset not meeting criteria for persistence, and adolescent/adult-onset. We tested associations between asthma and leukocyte telomere length using regression models. We tested for confounding of asthma-leukocyte telomere length associations using covariate adjustment. We tested serum C-reactive protein and white blood cell counts as potential mediators of asthma-leukocyte telomere length associations. Measurements and Main Results: Study members with life-course-persistent asthma had shorter leukocyte telomere length as compared with sex- and age-matched peers with no reported asthma. In contrast, leukocyte telomere length in study members with childhood-onset and adolescent/adult-onset asthma was not different from leukocyte telomere length in peers with no reported asthma. Adjustment for life histories of obesity and smoking did not change results. Study members with life-course-persistent asthma had elevated blood eosinophil counts. Blood eosinophil count mediated 29% of the life-course-persistent asthma-leukocyte telomere length association. Conclusions: Life-course-persistent asthma is related to a proposed biomarker of accelerated aging, possibly via systemic eosinophilic inflammation. Life histories of asthma can inform studies of aging. PMID:24956257

  8. Effects of eugenol on polymorphonuclear cell migration and chemiluminescence.

    PubMed

    Fotos, P G; Woolverton, C J; Van Dyke, K; Powell, R L

    1987-03-01

    In this study, the effects of eugenol on human polymorphonuclear (PMN) cell migration and chemiluminescence were examined in vitro. Utilizing zymosan-activated serum or crude Bacteroides sonicate fractions as chemotractants, we found that eugenol inhibits PMN migration at 6.6 X 10(-2) to 6.6 X 10(-5) mol/L (P less than 0.05). Also, similar effects were observed in PMNs pre-incubated in eugenol. Regardless of concentration, eugenol was not found to induce chemotaxis of PMNs. An examination of PMN membrane activation through chemiluminescence gave results consistent with the chemotaxis data, demonstrating a decrease in light emission at concentrations as low as 6.6 X 10(-6) mol/L (P less than 0.05). In view of these data, the potential effect of eugenol on in vivo (sulcular or periapical) PMN function deserves further study. PMID:3475310

  9. Decreased levels of peripheral leukocytes following sodium selenite treatment in female mice

    SciTech Connect

    Hogan, G.R.

    1986-08-01

    Selenium is known to be an essential micronutrient to a number of animal species (Schwarz 1961). Above its trace levels, however, selenium accumulation has long been known to induce deleterious conditions in domestic animals and in humans. Selenium has been reported to induce a hemolytic anemia, alter the configuration of plasma protein, reduce the synthesis of hemoglobin, depress packed red blood cell volume, and accumulate in renal and hepatic tissues. There are substantial data in regard to selenium effects on the erythrocyte component of peripheral blood, but there is an obvious deficiency in such information concerning the effects of selenium on the leukocyte component of blood. The major purpose of this investigation is to focus upon the effects of selenium on formed elements of the blood, and specifically, the leukocytes. Following three separate treatments of selenium, the number and class of peripheral leukocytes were determined as a function of time following administration of the selenium salt.

  10. The presence and persistence of Streptococcus uberis infections in the bovine udder and its leukocytic reaction 

    E-print Network

    Rahman, Mohammad Ataur

    1969-01-01

    of the cardinal characters of Str. uberis, Host of the Str. uberis are u-hemolytic and some are non-hemolytic. Based on such tests as hydrolysis of sodium-hippurate, splitting of esculin and hemolysis on blood agar, it is possible to distinguish differentiate... leukocytic counts. Table 4 shows the leukocytic reaction of the quarters infected with Str. uberis, Four quarters infected with hemolytic staphylococci gave high leukocytic counts. They were constant ranging from 570, 000 to 1. 5 million or more cells ir...

  11. A new insight into phagocytosis of apoptotic cells: proteolytic enzymes divert the recognition and clearance of polymorphonuclear leukocytes by macrophages

    Microsoft Academic Search

    K Guzik; M Bzowska; J Smagur; O Krupa; M Sieprawska; J Travis; J Potempa

    2007-01-01

    The recognition of phosphatidylserine (PS) on the surface of any apoptotic cell is considered to be a key event for its clearance. We challenge this concept by showing that pretreatment of neutrophils with either host or bacterial protease affects their uptake by human monocyte-derived macrophages without having an effect on cell-surface PS presentation. Specifically, whereas preincubation of apoptotic neutrophils with

  12. Biomaterials differentially regulate Src kinases and phosphoinositide 3-kinase-? in polymorphonuclear leukocyte primary and tertiary granule release.

    PubMed

    Cohen, Hannah Caitlin; Frost, Dustin C; Lieberthal, Tyler Jacob; Li, Lingjun; Kao, W John

    2015-05-01

    In the foreign body response, infiltrating PMNs exocytose granule subsets to influence subsequent downstream inflammatory and wound healing events. In previous studies, we found that PMNs cultured on poly(ethylene glycol) (PEG)-containing hydrogels (i.e., PEG and gelatin + PEG hydrogels) had enhanced primary granule release, yet similar tertiary granule release compared with PMNs cultured on polydimethylsiloxane or tissue culture polystyrene. PMN primary granules contain microbicidal proteins and proteases, which can potentially injure bystander cells, degrade the extracellular matrix, and promote inflammation. Here, we sought to understand the mechanism of the enhanced primary granule release from PMNs on PEG hydrogels. We found that primary granule release from PMNs on PEG hydrogels was adhesion mediated and involved Src family kinases and PI3K-?. The addition of gelatin to PEG hydrogels did not further enhance PMN primary granule release. Using stable-isotope dimethyl labeling-based shotgun proteomics, we identified many serum proteins - including Ig gamma constant chain region proteins and alpha-1-acid glycoprotein 1 - that were absorbed/adsorbed in higher quantities on PEG hydrogels than on TCPS, and may be involved in mediating PMN primary granule release. Ultimately, this mechanistic knowledge can be used to direct inflammation and wound healing following biomaterial implantation to promote a more favorable healing response. PMID:25736495

  13. Effects of Prostaglandins, Epinephrine and NaF on Human Leukocyte, Platelet and Liver Adenyl Cyclase

    Microsoft Academic Search

    ROBERT E. SCore

    1970-01-01

    Circulating leukocytes and platelets were isolated from hepaninized venous blood samples of 20 healthy males. These samples were immediately placed in siliconized containers and were diluted with an equal volume of three per cent dextran in 0.9 per cent saline exactly as described by Skoog and Beck.7 Following agglutination and sedimentation of the erythrocytic elements, the leukocyte-platelet-nich supernatant was decanted

  14. Absence of the Endothelial Oxidase AOC3 Leads to Abnormal Leukocyte Traffic In Vivo

    Microsoft Academic Search

    Craig M. Stolen; Fumiko Marttila-Ichihara; Kaisa Koskinen; Gennady G. Yegutkin; Raisa Turja; Petri Bono; Mikael Skurnik; Arno Hänninen; Sirpa Jalkanen; Marko Salmi

    2005-01-01

    Leukocyte migration from the blood to tissues is a prerequisite for normal immune responses. We produced mice deficient in an endothelial cell-surface oxidase (amine oxidase, copper containing-3 [AOC3], also known as vascular adhesion protein-1 [VAP-1]) and found that this enzyme is needed for leukocyte extravasation in vivo. Real-time imaging shows that AOC3 mediates slow rolling, firm adhesion, and transmigration of

  15. Arachidonic acid metabolism in the platelets and neutrophils of diabetic rabbit and human subjects

    Microsoft Academic Search

    Greco

    1985-01-01

    An alteration of arachidonic acid metabolism to prostaglandins and leukotrienes from platelets and polymorphonuclear leukocytes respectively is evident in subjects with diabetes mellitus. There is evidence of altered platelet\\/vascular wall interactions in diabetes mellitus and evidence that polymorphonuclear leukocytes influence the vascular walls. Theories on the pathogenesis of atherosclerosis include both blood cells. Platelet hypersensitivity is evident in those platelets

  16. The Effects of Psychological Stress on Leukocyte Subset Distribution in Humans: Evidence of Immune Activation

    Microsoft Academic Search

    Michael Maes; Dirk R. Van Bockstaele; Ann Van Gastel; Cai Song; Chris Schotte; Hugo Neels; Ingrid DeMeester; Simon Scharpe; Aleksander Janca

    1999-01-01

    The aim of the present study was to examine the effects of academic examination stress on leukocyte subset distribution in university students. Thirty-eight university students had repeated blood collections for white blood cell differentiation and flow cytometric assay of lymphocytic subsets a few weeks before and after (i.e. two baseline conditions) as well as the day before a difficult academic

  17. The effect of citrus-derived oil on bovine blood neutrophil function and gene expression in vitro.

    PubMed

    Garcia, M; Elsasser, T H; Biswas, D; Moyes, K M

    2015-02-01

    Research on the use of natural products to treat or prevent microbial invasion as alternatives to antibiotic use is growing. Polymorphonuclear leukocytes (PMNL) play a vital role with regard to the innate immune response that affects severity or duration of mastitis. To our knowledge, effect of cold-pressed terpeneless Valencia orange oil (TCO) on bovine PMNL function has not been elucidated. Therefore, the objective of this study was to investigate the effect of TCO on bovine blood PMNL chemotaxis and phagocytosis capabilities and the expression of genes involved in inflammatory response in vitro. Polymorphonuclear leukocytes were isolated from jugular blood of 12 Holstein cows in mid-lactation and were incubated with 0.0 or 0.01% TCO for 120min at 37°C and 5% CO2, and phagocytosis (2×10(6) PMNL) and chemotaxis (6×10(6) PMNL) assays were then performed in vitro. For gene expression, RNA was extracted from incubated PMNL (6×10(6) PMNL), and gene expression was analyzed using quantitative PCR. The supernatant was stored at -80°C for analysis of tumor necrosis factor-?. Data were analyzed using a general linear mixed model with cow and treatment (i.e., control or TCO) in the model statement. In vitro supplementation of 0.01% of TCO increased the chemotactic ability to IL-8 by 47%; however, migration of PMNL to complement 5a was not altered. Treatment did not affect the production of tumor necrosis factor-? by PMNL. Expression of proinflammatory genes (i.e., SELL, TLR4, IRAK1, TRAF6, and LYZ) coding for proteins was not altered by incubation of PMNL with TCO. However, downregulation of TLR2 [fold change (FC=treatment/control)=-2.14], NFKBIA (FC=1.82), IL1B (FC=-2.16), TNFA (FC=-9.43), and SOD2 (FC=-1.57) was observed for PMNL incubated with TCO when compared with controls. Interestingly, expression of IL10, a well-known antiinflammatory cytokine, was also downregulated (FC=-3.78), whereas expression of IL8 (FC=1.93), a gene coding for the cytokine IL-8 known for its chemotactic function, tended to be upregulated in PMNL incubated with TCO. Incubation of PMNL with TCO enhanced PMNL chemotaxis in vitro. The expression of genes involved in the inflammatory response was primarily downregulated. Results showed that 0.01% TCO did not impair the function of PMNL in vitro. Future studies investigating the use of TCO as an alternative therapy for treatment of mastitis, including dose and duration, for cows during lactation are warranted. PMID:25434342

  18. AVIAN POLYMORPHONUCLEAR CELLS CONTRIBUTE TO A DIFFERENTIAL INNATE IMMUNE RESPONSE IN GENETICALLY DEFINED CHICKENS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Heterophils, the primary polymorphonuclear cell (PMN) in chickens, are the avian counterpart to mammalian neutrophils. Heterophils modulate acute innate responses through phagocytosis, respiratory burst, and degranulation. We have been characterizing the heterophil-mediated innate immune response ...

  19. Integrin Regulation during Leukocyte Recruitment.

    PubMed

    Herter, Jan; Zarbock, Alexander

    2013-05-01

    Integrins are recognized as vital players in leukocyte recruitment. Integrin malfunction causes severe disease patterns characterized by the inability to fight pathogens. Although inflammatory reactions are beneficial and necessary for host defense, these reactions have to be controlled to prevent tissue destruction and harmful sequelae. In this review, we discuss the different signaling pathways leading to the change of integrin adhesiveness in neutrophils, monocytes, and lymphocytes. We thereby focus on the importance of integrin activation for the different steps of the leukocyte recruitment cascade, including rolling, adhesion, postadhesion strengthening, intravascular crawling, and transmigration, as each step necessitates the proper functioning of a distinct set of integrin molecules that has to be activated specifically. Additionally, we discuss endogenous mechanisms that balance and counteract integrin activation and limit leukocyte recruitment at the site of inflammation. Further insight into these complex mechanisms may provide new approaches for developing new anti-inflammatory therapies. PMID:23606722

  20. Effect of small cationic leukocyte peptides (defensins) on the permeability barrier of the outer membrane.

    PubMed Central

    Viljanen, P; Koski, P; Vaara, M

    1988-01-01

    Defensins are small cationic antibacterial peptides that are abundant in polymorphonuclear leukocytes from human and other sources (T. Ganz, M. Selsted, D. Szklarek, S. Harwig, K. Daher, D. F. Bainton, and R. J. Lehrer, J. Clin. Invest. 76:1427-1435, 1985). We studied whether subinhibitory concentrations of defensins increase the outer membrane (OM) permeability of Escherichia coli, Salmonella typhimurium, and Pseudomonas aeruginosa to hydrophobic probes, as do many other polycations that have been studied previously. Throughout the study, we used polymyxin B nonapeptide (PMBN) as a reference peptide. PMBN has a known potent OM permeability-increasing action. As a sharp contrast to PMBN, subinhibitory concentrations of defensins did not permeabilize (or, under some test conditions, permeabilized very slightly) the OM to the probes that were used (rifampin and Triton X-100). At bacteriostatic or bactericidal defensin concentrations, some degree of synergism with rifampin was seen. Images PMID:3137167

  1. METABOLIC AND MORPHOLOGICAL OBSERVATIONS ON THE EFFECT OF SURFACE-ACTIVE AGENTS ON LEUKOCYTES

    PubMed Central

    Graham, R. C.; Karnovsky, M. J.; Shafer, A. W.; Glass, E. A.; Karnovsky, Manfred L.

    1967-01-01

    Morphological and metabolic observations have been made on the effects of endotoxin, deoxycholate, and digitonin (at less than 50 µg/ml) on polymorphonuclear leukocytes and mononuclear cells. The agents stimulate the respiration and glucose oxidation of these cells in a manner similar to that seen during phagocytosis. Electron microscopy revealed no morphological changes with the first two agents, but dramatic membrane changes were seen in the case of digitonin. Here tubular projections of characteristic size and shape formed on and split off the membrane. All the agents stimulated uptake of inulin, but efforts to demonstrate increased pinocytosis by electron microscopy have not so far succeeded, probably due to limitations in present experimental techniques. PMID:6034482

  2. Hematologically important mutations: leukocyte adhesion deficiency (first update).

    PubMed

    van de Vijver, Edith; Maddalena, Anne; Sanal, Özden; Holland, Steven M; Uzel, Gulbu; Madkaikar, Manisha; de Boer, Martin; van Leeuwen, Karin; Köker, M Yavuz; Parvaneh, Nima; Fischer, Alain; Law, S K Alex; Klein, Nigel; Tezcan, F Ilhan; Unal, Ekrem; Patiroglu, Turkan; Belohradsky, Bernd H; Schwartz, Klaus; Somech, Raz; Kuijpers, Taco W; Roos, Dirk

    2012-01-15

    Leukocyte adhesion deficiency (LAD) is an immunodeficiency caused by defects in the adhesion of leukocytes (especially neutrophils) to the blood vessel wall. As a result, patients with LAD suffer from severe bacterial infections and impaired wound healing, accompanied by neutrophilia. In LAD-I, mutations are found in ITGB2, the gene that encodes the ? subunit of the ?(2) integrins. This syndrome is characterized directly after birth by delayed separation of the umbilical cord. In the rare LAD-II disease, the fucosylation of selectin ligands is disturbed, caused by mutations in SLC35C1, the gene that encodes a GDP-fucose transporter of the Golgi system. LAD-II patients lack the H and Lewis Le(a) and Le(b) blood group antigens. Finally, in LAD-III (also called LAD-I/variant) the conformational activation of the hematopoietically expressed ? integrins is disturbed, leading to leukocyte and platelet dysfunction. This last syndrome is caused by mutations in FERMT3, encoding the kindlin-3 protein in all blood cells that is involved in the regulation of ? integrin conformation. PMID:22134107

  3. Cytochemical Leukocyte Reactions in Normal Newborn Infants

    Microsoft Academic Search

    J. Corberand; J. Pris; C. Régnier

    1973-01-01

    The scores of five cytochemical leukocyte reactions (myeloperoxydases, leukocyte alkaline phosphatases, nonspecific esterases, cell polysaccharides and Soudan black) were established in a group of 100 healthy neonates from normal pregnancies. Comparisons between infant and adult findings show that leukocyte alkaline phosphatase activity is distinctly higher in infants whereas that of mieloperoxydases and of lymphocyte cell polysaccharides is lower. The interpretation

  4. Tumor necrosis factor and interferon-gamma induce distinct patterns of endothelial activation and associated leukocyte accumulation in skin of Papio anubis.

    PubMed Central

    Munro, J. M.; Pober, J. S.; Cotran, R. S.

    1989-01-01

    Recombinant human interferon (IFN)-gamma (2 X 10(4) or 2 X 10(5) U), tumor necrosis factor (TNF, 10(4) or 10(5) U), or both were injected intracutaneously into baboons (Papio anubis), and biopsies were examined at various intervals for evidence of altered endothelial cell antigen expression, endothelial morphology, and leukocyte infiltration. IFN-gamma induced increased binding of anti-HLA-DP mAb by 24 hours and a mild-to-moderate accumulation of mononuclear cells. TNF induced increased binding of anti-endothelial leukocyte adhesion molecule (ELAM)-1 mAb by 2 hours that was associated with polymorphonuclear leukocyte accumulation, and increased binding of anti-intercellular adhesion molecule (ICAM)-1 mAb by 9 hours that was associated with the onset of progressive mononuclear leukocyte accumulation. TNF also caused endothelial cell hypertrophy and increased vascular permeability. The combination of IFN-gamma and TNF induced a set of changes that qualitatively resemble those of a delayed hypersensitivity reaction to simian agent 8 envelope antigen. These findings are consistent with the concept that cytokine-activated endothelium plays an important role in the adhesion and subsequent extravasation of leukocytes during immune inflammation. Images Figure 1 Figure 3 Figure 4 Figure 5 Figure 6 PMID:2505619

  5. Channel catfish (Ictalurus punctatus) leukocytes express estrogen receptor isoforms ER? and ER?2 and are functionally modulated by estrogens.

    PubMed

    Iwanowicz, Luke R; Stafford, James L; Patińo, Reynaldo; Bengten, Eva; Miller, Norman W; Blazer, Vicki S

    2014-09-01

    Estrogens are recognized as modulators of immune responses in mammals and teleosts. While it is known that the effects of estrogens are mediated via leukocyte-specific estrogen receptors (ERs) in humans and mice, leucocyte-specific estrogen receptor expression and the effects of estrogens on this cell population is less explored and poorly understood in teleosts. Here in, we verify that channel catfish (Ictalurus punctaus) leukocytes express ER? and ER?2. Transcripts of these isoforms were detected in tissue-associated leukocyte populations by PCR, but ER?2 was rarely detected in PBLs. Expression of these receptors was temporally regulated in PBLs following polyclonal activation by concanavalin A, lipopolysaccharide or alloantigen based on evaluation by quantitative and end-point PCR. Examination of long-term leukocyte cell lines demonstrated that these receptors are differentially expressed depending on leukocyte lineage and phenotype. Expression of ERs was also temporally dynamic in some leukocyte lineages and may reflect stage of cell maturity. Estrogens affect the responsiveness of channel catfish peripheral blood leukocytes (PBLs) to mitogens in vitro. Similarly, bactericidal activity and phorbol 12-myristate 13-acetate induced respiratory burst was modulated by 17?-estradiol. These actions were blocked by the pure ER antagonist ICI 182780 indicating that response is, in part, mediated via ER?. In summary, estrogen receptors are expressed in channel catfish leukocytes and participate in the regulation of the immune response. This is the first time leukocyte lineage expression has been reported in teleost cell lines. PMID:24973517

  6. Observing a fictitious stressful event: haematological changes, including circulating leukocyte activation.

    PubMed

    Mian, Rubina; Shelton-Rayner, Graham; Harkin, Brendan; Williams, Paul

    2003-03-01

    The aim of this study was to assess the effect of watching a psychological stressful event on the activation of leukocytes in healthy human volunteers. Blood samples were obtained from 32 healthy male and female subjects aged between 20 and 26 years before, during and after either watching an 83-minute horror film that none of the subjects had previously seen (The Texas Chainsaw Massacre, 1974) or by sitting quietly in a room (control group). Total differential cell counts, leukocyte activation as measured by the nitroblue tetrazolium (NBT) test, heart rate and blood pressure (BP) measurements were taken at defined time points. There were significant increases in peripheral circulating leukocytes, the number of activated circulating leukocytes, haemoglobin (Hb) concentration and haematocrit (Hct) in response to the stressor. These were accompanied by significant increases in heart rate, systolic and diastolic BP (P<0.05 from baseline). This is the first reported study on the effects of observing a psychologically stressful, albeit fictitious event on circulating leukocyte numbers and the state of leukocyte activation as determined by the nitrotetrazolium test. PMID:12637206

  7. Leukocyte integrins: role in leukocyte recruitment and as therapeutic targets in inflammatory disease.

    PubMed

    Mitroulis, Ioannis; Alexaki, Vasileia I; Kourtzelis, Ioannis; Ziogas, Athanassios; Hajishengallis, George; Chavakis, Triantafyllos

    2015-03-01

    Infection or sterile inflammation triggers site-specific attraction of leukocytes. Leukocyte recruitment is a process comprising several steps orchestrated by adhesion molecules, chemokines, cytokines and endogenous regulatory molecules. Distinct adhesive interactions between endothelial cells and leukocytes and signaling mechanisms contribute to the temporal and spatial fine-tuning of the leukocyte adhesion cascade. Central players in the leukocyte adhesion cascade include the leukocyte adhesion receptors of the ?2-integrin family, such as the ?L?2 and ?M?2 integrins, or of the ?1-integrin family, such as the ?4?1-integrin. Given the central involvement of leukocyte recruitment in different inflammatory and autoimmune diseases, the leukocyte adhesion cascade in general, and leukocyte integrins in particular, represent key therapeutic targets. In this context, the present review focuses on the role of leukocyte integrins in the leukocyte adhesion cascade. Experimental evidence that has implicated leukocyte integrins as targets in animal models of inflammatory disorders, such as experimental autoimmune encephalomyelitis, psoriasis, inflammatory bone loss and inflammatory bowel disease as well as preclinical and clinical therapeutic applications of antibodies that target leukocyte integrins in various inflammatory disorders are presented. Finally, we review recent findings on endogenous inhibitors that modify leukocyte integrin function, which could emerge as promising therapeutic targets. PMID:25448040

  8. Derivation of Cinnamon Blocks Leukocyte Attachment by Interacting with Sialosides

    PubMed Central

    Lin, Wei-Ling; Guu, Shih-Yun; Tsai, Chan-Chuan; Prakash, Ekambaranellore; Viswaraman, Mohan; Chen, Hsing-Bao; Chang, Chuan-Fa

    2015-01-01

    Molecules derived from cinnamon have demonstrated diverse pharmacological activities against infectious pathogens, diabetes and inflammatory diseases. This study aims to evaluate the effect of the cinnamon-derived molecule IND02 on the adhesion of leukocytes to host cells. The anti-inflammatory ability of IND02, a pentameric procyanidin type A polyphenol polymer isolated from cinnamon alcohol extract, was examined. Pretreatment with IND02 significantly reduced the attachment of THP-1 cells or neutrophils to TNF-?-activated HUVECs or E-selectin/ICAM-1, respectively. IND02 also reduced the binding of E-, L- and P-selectins with sialosides. Furthermore, IND02 could agglutinate human red blood cells (RBC), and the agglutination could be disrupted by sialylated glycoprotein. Our findings demonstrate that IND02, a cinnamon-derived compound, can interact with sialosides and block the binding of selectins and leukocytes with sialic acids. PMID:26076445

  9. Evaluation of human polymorphonuclear behavior on textured titanium and calcium-phosphate coated surfaces.

    PubMed

    Moura, Camilla C G; Machado, Juliana R; Silva, Marcos V; Rodrigues, Denise B R; Zanetta-Barbosa, Darceny; Jimbo, Ryo; Tovar, Nick; Coelho, Paulo G

    2013-06-01

    Few studies have evaluated the effects of titanium (Ti) surface modifications on polymorphonuclear neutrophils (PMNs). Human PMNs' viability and release of key mediators-such as IL1?, IL6, TNF?, IL12, IL10, IL4, TGF?1, IL8, IP-10, and Mig-were evaluated on three different Ti surface treatments: (1) machined Ti; (2) alumina-blasted and acid-etched Ti (AB/AE); and (3) calcium phosphate coating of 300-500 nm by ion beam onto the AB/AE Ti surface (CaP). A polystyrene surface was used as a negative control. The PMNs were purified from whole human blood and cultured for 6 h. Cell viability was determined by flow cytometry, and the supernatant was evaluated to determine the levels of cytokines and chemokines. Results showed that the percentage of viable cells was significantly lower on the CaP surface compared to the control (p < 0.05) relative to the other groups. No differences in the levels of IL8, MIG, and IP10 were detected between groups. Significantly higher levels of IL1? (p = 0.046) and TNF? (p = 0.016) were detected for the CaP surfaces compared to AB/AE surface only. The levels of IL4, IL10, and TGF?1 secreted from the PMNs in the CaP group were significantly lower than in the control and machined groups (p < 0.05) that were statistically comparable to AB/AE. Overall, the addition of a thin CaP coating to the AB/AE Ti surface influenced the secretion profile of pro-inflammatory cytokines due to the higher release of pro-inflammatory cytokines (IL1? and TNF?) on these surfaces. PMID:23598427

  10. Expression of CD11b and CD18 on polymorphonuclear neutrophils stimulated with interleukin-2

    PubMed Central

    Abdel-Salam, Bahaa K.A.

    2014-01-01

    Background Interleukin-2 (IL-2) is a lymphocyte-activating and growth-promoting factor, and has been widely studied on T-cells and NK-cells. However, the interaction of polymorphonuclear neutrophils (PMNs) with IL-2 is poorly studied and thus, this study aimed at defining IL-2 participation in the expression of CD11b and CD18 on PMNs. Material and methods PMNs were isolated from heparinized whole blood of healthy donors. Purified cells were incubated with IL-2 (10 ng/ml) for 24 hours at 37°C in a humidified incubator with 5% CO2. After 24 hours’ incubation, surface molecules (CD11b and CD18) were measured by flow cytometry. Results Interestingly, the antibodies of IL-2R? chain (CD122-FITC) were found in all observed cells. The induction of CD11b mean fluorescence intensity (MFI) in highly purified PMNs stimulated with IL-2 was clearly increased recording 43% in comparison to the freshly isolated PMNs and the un-stimulated PMNs which were found to be 23% and 28% of CD11b, respectively. Furthermore, flow cytometry analysis demonstrated that the highly purified PMNs exposed to IL-2 showed an increase in CD18 MFI, recording 47% with respect to that of the freshly isolated PMNs and PMNs cultured with the medium alone which showed a small amount of 38% and 27%, respectively. Conclusions Results demonstrated that CD11b and CD18 had been acquired on the surface of the IL-2-in vitro-activated PMNs. These findings indicated that IL-2 may play a crucial role in PMNs migration.

  11. Effect of acetaminophen on the leukocyte-labeling efficiency of indium oxine In 111

    SciTech Connect

    Augustine, S.C.; Schmelter, R.F.; Nelson, K.L.; Petersen, R.J.; Qualfe, M.A.

    1983-11-01

    The effect of acetaminophen on the labeling efficiency of leukocytes with indium oxine In 111 was studied. A blood sample was obtained from eight healthy men before and after they received acetaminophen 650 mg every four hours for 24 hours. After dividing the plasma from each sample into three portions, leukocytes were separated and labeled with indium oxine In 111. In an in vitro study, 200 ml of blood was obtained from one of the men, and the plasma was separated into four portions. Acetaminophen in 95% ethanol was added to three of the plasma fractions to produce acetaminophen concentrations of 4, 20, and 100 micrograms/ml; ethanol was added to the fourth fraction as a control. Each plasma fraction was then subdivided into three aliquots, and leukocytes were labeled as in the in vivo study. Mean leukocyte labeling efficiencies in both studies were calculated from the ratios of leukocyte radioactivity to initial radioactivity in the samples, expressed as percentages. Leukocyte labeling efficiencies before acetaminophen administration ranged from 79 to 85%; after administration, labeling efficiencies ranged from 70 to 87%. No significant differences in mean labeling efficiency before and after acetaminophen administration were noted in any of the subjects. Leukocyte labeling efficiencies in all in vitro plasma fractions were reduced, ranging from 54 to 63%, but no significant differences in labeling efficiency between any of the plasma fractions were found. Using the labeling procedures in this study, exposure of leukocytes from healthy men to acetaminophen in vivo or in vitro does not affect labeling efficiency with indium oxine In 111.

  12. Oxaceprol, an atypical inhibitor of inflammation, reduces leukocyte adherence in mouse antigen-induced arthritis.

    PubMed

    Veihelmann, A; Hofbauer, A; Refior, H J; Messmer, K

    2001-06-01

    Oxaceprol (N-acetyl-L-hydroxyproline), an atypical inhibitor of inflammation, is an established drug forjoint disease without serious side-effects. Recent studies have emphasized that oxaceprol has an effect on the microcirculation. Since the exact mechanism of action remains unclear, the aim of our study was to investigate the leukocyte-endothelial cell interactions in oxaceprol-treated mice with antigen-induced arthritis (AiA) using intravital microscopy. In our study, Balb/c mice were allocated to 4 groups (n 7, 8, 8, 8): 2 control groups with saline or oxaceprol and 2 groups of arthritic animals which received saline or oxaceprol (100 mg/kg twice a day intraperitoneally). The severity of arthritis was quantified by the transverse knee joint diameter. For the intravital fluorescence microscopy measurements on day 10 after inducing arthritis, the patella tendon was partily resected to visualize the intraarticular synovial tissue of the knee joint. The number of rolling and adherent leukocytes as well as RBC velocity and functional capillary density (FCD) were quantified in synovial microvessels. Furthermore, leukocyte infiltration was determined in the histological sections with an established score. No significant changes in mean arterial blood pressure or functional capillary density were found in any of the groups. However, the leukocyte rolling fraction and number of leukocytes adherent to the endothelium were increased in postcapillary venules of the synovium in arthritic animals (0.16 to 0.31, 78 cells/mm2 to 220 cells/mm2). In animals with AiA treated with oxaceprol, leukocyte adherence and swelling were significantly reduced in comparison to the arthritic animals treated with saline. Furthermore, the histological score showed less leukocyte infiltration in the oxaceprol treated arthritic animals. Thus, oxaceprol reduces leukocyte adherence in vivo and leukocyte infiltration in mouse AiA, indicating an effect on synovial microcirculation. PMID:11480608

  13. Leukocyte Recruitment in Inflammation: Basic Concepts and New Mechanistic Insights Based on New Models and Microscopic Imaging Technologies

    PubMed Central

    Leick, Marion; Azcutia, Veronica; Newton, Gail; Luscinskas, Francis W.

    2014-01-01

    The immune cell system is a critical component of host defense. Recruitment of immune cells to sites of infection, immune reaction, or injury is complex and involves coordinated adhesive interactions between the leukocyte and the endothelial cell monolayer that lines blood vessels. This article will review basic mechanisms in the recruitment of leukocytes to tissues, and then selectively review new concepts that are emerging based on advances in live cell imaging microscopy and mouse strains. These emerging concepts are altering the conventional paradigms of inflammatory leukocyte recruitment established in the early 1990’s. Indeed, recent publications have identified previously unrecognized contributions from pericytes and interstitial leukocytes and their secreted products that guide leukocytes to their targets. It is also notable that investigators have begun to design organs on a chip. Recent reports indicate that this avenue of research holds much promise. PMID:24562377

  14. Chemotactic factor influences on the aggregation, swelling, and foreign surface adhesiveness of human leukocytes.

    PubMed Central

    O'Flaherty, J. T.; Kreutzer, D. L.; Ward, P. A.

    1978-01-01

    Chemotactic factors have been shown to induce aggregation and cellular swelling of rabbit polymorphonuclear neutrophils (PMN) obtained from the peritoneum. We examined the ability of the chemotactic fragment of C5 and the synthetic chemotactic tripeptide formyl-methionyl-leucyl-phenylalanine to induce these changes in various preparations of human leukocytes. We found that these factors did induce dextran-sedimented leukocytes and Ficoll-Hypaque-isolated PMN to aggregate and swell. Compared with rabbit peritoneal PMN, however, human PMN responded with more prominent swelling but with less prominent aggregation. Also unlike rabbit peritoneal PMN, human PMN adhered spontaneously to plastic surfaces; the chemotactic factors enhanced this adherence. Certain similarities between the responses of these two cell types were evident: in both rabbit peritoneal and isolated human peripheral PMN, the aggregates had a short life span in the fluid phase; in both, the number of aggregates formed was proportional to the log10 of the PMN concentration; and, in both, the chemotactic activity of the reagents paralleled their aggregating activity. In the system employed, lymphocytes were unresponsive to the chemotactic factors. Ficoll-Hypaque-isolated mononoclear cells (containing varying proportions of monocytes and lymphocytes) were responsive, indicating that human monocytes behave in a manner similar to the human PMN. The results suggest that chemotactic factors induce responsive cells to develop a hyperadherent cytoplasmic membrane. Aggregation and increased adhesiveness to plastic surfaces may reflect this induction. PMID:564610

  15. The leukocyte response to fluid?stress

    PubMed Central

    Moazzam, Fariborz; DeLano, Frank A.; Zweifach, Benjamin W.; Schmid-Schönbein, Geert W.

    1997-01-01

    Leukocyte migration from a hemopoietic pool across marrow endothelium requires active pseudopod formation and adhesion. Leukocytes rarely show pseudopod formation while in circulation. At question then is the mechanism that serves to minimize leukocyte pseudopod formation in the circulation. We tested the hypothesis that fluid shear stress acts to prevent pseudopod formation. When individual human leukocytes (neutrophils, monocytes) spreading on glass surfaces in vitro were subjected to fluid shear stress (?1 dyn/cm2), an instantaneous retraction of pseudopods was observed. Removal of the fluid shear stress in turn led to the return of pseudopod projection and cell spreading. When steady shear stress was prolonged over several minutes, leukocyte swelling occurs together with an enhanced random motion of cytoplasmic granules and a reduction of cytoplasmic stiffness. The response to shear stress could be suppressed by K+ channel blockers and chelation of external Ca2+. In rat mesentery microvessels after occlusion, circulating leukocytes project pseudopods in free suspension or when attached to the endothelium, even though immediately after occlusion only few pseudopods were present. When flow was restored, pseudopods on adhering leukocytes were retracted and then the cells began to roll and detach from the endothelium. In conclusion, plasma shear stress in the circulation serves to reduce pseudopod projection and adhesion of circulating leukocytes and vice versa reduction of shear stress leads to pseudopod projection and spreading of leukocytes on the endothelium. PMID:9144238

  16. Isolation and Analysis of Brain-sequestered Leukocytes from Plasmodium berghei ANKA-infected Mice

    PubMed Central

    Ryg-Cornejo, Victoria; Ioannidis, Lisa J.; Hansen, Diana S.

    2013-01-01

    We describe a method for isolation and characterization of adherent inflammatory cells from brain blood vessels of P. berghei ANKA-infected mice. Infection of susceptible mouse-strains with this parasite strain results in the induction of experimental cerebral malaria, a neurologic syndrome that recapitulates certain important aspects of Plasmodium falciparum-mediated severe malaria in humans 1,2 . Mature forms of blood-stage malaria express parasitic proteins on the surface of the infected erythrocyte, which allows them to bind to vascular endothelial cells. This process induces obstructions in blood flow, resulting in hypoxia and haemorrhages 3 and also stimulates the recruitment of inflammatory leukocytes to the site of parasite sequestration. Unlike other infections, i.e neutrotopic viruses4-6, both malaria-parasitized red blood cells (pRBC) as well as associated inflammatory leukocytes remain sequestered within blood vessels rather than infiltrating the brain parenchyma. Thus to avoid contamination of sequestered leukocytes with non-inflammatory circulating cells, extensive intracardial perfusion of infected-mice prior to organ extraction and tissue processing is required in this procedure to remove the blood compartment. After perfusion, brains are harvested and dissected in small pieces. The tissue structure is further disrupted by enzymatic treatment with Collagenase D and DNAse I. The resulting brain homogenate is then centrifuged on a Percoll gradient that allows separation of brain-sequestered leukocytes (BSL) from myelin and other tissue debris. Isolated cells are then washed, counted using a hemocytometer and stained with fluorescent antibodies for subsequent analysis by flow cytometry. This procedure allows comprehensive phenotypic characterization of inflammatory leukocytes migrating to the brain in response to various stimuli, including stroke as well as viral or parasitic infections. The method also provides a useful tool for assessment of novel anti-inflammatory treatments in pre-clinical animal models. PMID:23329000

  17. Cell-stiffness-induced mechanosignaling - a key driver of leukocyte transendothelial migration.

    PubMed

    Schaefer, Antje; Hordijk, Peter L

    2015-07-01

    The breaching of cellular and structural barriers by migrating cells is a driving factor in development, inflammation and tumor cell metastasis. One of the most extensively studied examples is the extravasation of activated leukocytes across the vascular endothelium, the inner lining of blood vessels. Each step of this leukocyte transendothelial migration (TEM) process is regulated by distinct endothelial adhesion receptors such as the intercellular adhesion molecule 1 (ICAM1). Adherent leukocytes exert force on these receptors, which sense mechanical cues and transform them into localized mechanosignaling in endothelial cells. In turn, the function of the mechanoreceptors is controlled by the stiffness of the endothelial cells and of the underlying substrate representing a positive-feedback loop. In this Commentary, we focus on the mechanotransduction in leukocytes and endothelial cells, which is induced in response to variations in substrate stiffness. Recent studies have described the first key proteins involved in these mechanosensitive events, allowing us to identify common regulatory mechanisms in both cell types. Finally, we discuss how endothelial cell stiffness controls the individual steps in the leukocyte TEM process. We identify endothelial cell stiffness as an important component, in addition to locally presented chemokines and adhesion receptors, which guides leukocytes to sites that permit TEM. PMID:26092932

  18. Further comparisons of endogenous pyrogens and leukocytic endogenous mediators.

    PubMed Central

    Kampschmidt, R F; Upchurch, H F; Worthington, M L

    1983-01-01

    It was recently shown (Murphy et al., Infect. Immun. 34:177-183), that rabbit macrophages produce two biochemically and immunologically distinct endogenous pyrogens. One of these has or copurifies with substances having a molecular weight of 13,000 and a pI of 7.3. This protein was produced by blood monocytes or inflammatory cells elicited in 16-h rabbit peritoneal exudates. These acute peritoneal exudates were produced by the intraperitoneal injection of large volumes of saline containing shellfish glycogen. When the leukocytes in these exudates were washed and incubated at 37 degrees C in saline, they released an endogenous pyrogen. The injection of this pyrogen into rabbits, rats, or mice caused the biological manifestations which have been attributed to leukocytic endogenous mediator. These effects were increases in blood neutrophils, the lowering of plasma iron and zinc levels, and the increased synthesis of the acute-phase proteins. The other rabbit endogenous pyrogen seems to be a family of proteins with isoelectric points between 4.5 and 5.0. These proteins are produced by macrophages in the lung, liver, or in chronic peritoneal exudates. In these experiments, the lower-isoelectric-point endogenous pyrogens were produced by macrophages from the peritoneal cavity of rabbits that had been injected 4 days earlier with 50 ml of light mineral oil. These rabbit pyrogens were found to have leukocytic endogenous mediator activity in mice but to be completely inactive in rats. When injected into rabbits, these proteins produced fever, lowered plasma iron, increased blood neutrophils, but failed to elevate plasma fibrinogen. PMID:6862633

  19. Production of immunoreactive polymorphonuclear leucocyte elastase in human breast cancer cells: possible role of polymorphonuclear leucocyte elastase in the progression of human breast cancer

    Microsoft Academic Search

    J-I Yamashita; M Ogawa; S Ikei; H Omachi; S-I Yamashita; T Saishoji; K Nomura; H Sato

    1994-01-01

    Breast cancer cells are known to express various proteolytic enzymes, which make them invasive and favour their dissemination to distant sites. However, it is unclear whether breast cancer cells have the ability to produce polymorphonuclear leucocyte elastase (PMN-E). We measured immunoreactive (ir) PMN-E content in the conditioned medium of two breast cancer cell lines, MCF-7 and ZR-75-1, and two normal

  20. 8Hydroxy2?Deoxyguanosine of Leukocyte DNA as a Marker of Oxidative Stress in Chronic Hemodialysis Patients

    Microsoft Academic Search

    Der-Cherng Tarng; Tung-Po Huang; Yau-Huei Wei; Tsung-Yun Liu; Haw-Wen Chen; Tzen Wen Chen; Wu-Chang Yang

    2000-01-01

    In contrast to proteins and lipids, oxidative damage to DNA has not been well studied in patients undergoing hemodialysis (HD). We hypothesized that phagocytes are activated after blood-membrane contact during HD, and oxidants from metabolic activation can damage leukocyte DNA. To test this hypothesis, the 8-hydroxy-2?-deoxyguanosine (8-OHdG) content of leukocyte DNA was measured by high-performance liquid chromatography electrochemical detection method

  1. Genetics Home Reference: Leukocyte adhesion deficiency type 1

    MedlinePLUS

    ... Genetic disorder catalog Conditions > Leukocyte adhesion deficiency type 1 On this page: Description Genetic changes Inheritance Diagnosis ... April 2014 What is leukocyte adhesion deficiency type 1? Leukocyte adhesion deficiency type 1 is a disorder ...

  2. Photoperiodic Regulation of Circulating Leukocytes in Juvenile Siberian Hamsters: Mediation by Melatonin and Testosterone

    Microsoft Academic Search

    Brian J. Prendergast; Andrew K. Hotchkiss; Randy J. Nelson

    2003-01-01

    The reproductive system of Siberian hamsters (Phodopus sungorus) undergoes rapid phenotypic responses to changes in day length that occur around the time of weaning. The present experiments tested whether the immune system of Siberian hamsters is similarly photoperiodicearly in life and whether photoperiodic changes in melatonin or gonadal hormone secretions mediate any such responses to day length. Circulating blood leukocyte

  3. Selective reduction of the interaction of magnetic nanoparticles with leukocytes and tumor cells by human plasma

    NASA Astrophysics Data System (ADS)

    Schwalbe, Manuela; Jörke, Cornelia; Buske, Norbert; Höffken, Klaus; Pachmann, Katharina; Clement, Joachim H.

    2005-05-01

    Carboxymethyl-dextran coated magnetic nanoparticles can interact with viable human cells. The interaction of the nanoparticles is cell-type specific. The addition of human plasma led to a dramatic reduction of magnetically separable leukocytes in comparison to tumor cells. We conclude that low plasma concentrations might support an efficient enrichment of circulating epithelial cells from the peripheral blood of tumor patients.

  4. Mechanics of Leukocyte Deformation and Adhesion to Endothelium in Shear Flow

    Microsoft Academic Search

    Cheng Dong; Jian Cao; Erika J. Struble; Herbert H. Lipowsky

    1999-01-01

    The mechanics of leukocyte [white blood cell (WBC)] deformation and adhesion to endothelial cells (EC) in shear flow has been investigated. Experimental data on transient WBC–EC adhesion were obtained from in vivo measurements. Microscopic images of WBC–EC contact during incipient WBC rolling revealed that for a given wall shear stress, the contact area increases with time as new bonds are

  5. The state diagram for cell adhesion under flow: Leukocyte rolling and firm adhesion

    E-print Network

    Tees, David F.J.

    method called ``Adhe- sive Dynamics,'' we have simulated the adhesion of a cell to a surface in flow an unbound blood cell to an adherent one in flow involves a number of steps: initial tethering, transientThe state diagram for cell adhesion under flow: Leukocyte rolling and firm adhesion Kai-Chien Chang

  6. Measurement of oxyradicals from leukocytes lodged in a microchannel array

    NASA Astrophysics Data System (ADS)

    Kikuchi, Yuji; Kikuchi, Hiroko E.

    2001-05-01

    The oxyradical production by leukocytes is crucial for killing invading bacteria, while it has been widely discussed as a tissue injuring factor. Despite such importance of the event, it is still unclear, probably because of lack of simple and reliable measurement methods, to what extent it varies among different subjects and either to what extent it is affected by environmental factors including diet. The present paper describes use of microfabricated channel arrays, that have been developed for use in studies of blood rheology, in the oxyradical measurement by chemiluminescence.

  7. Nanowire array chips for molecular typing of rare trafficking leukocytes with application to neurodegenerative pathology

    PubMed Central

    Kwak, Minsuk; Kim, Dong-Joo; Lee, Mi-Ri; Wu, Yu; Han, Lin; Lee, Sang-Kwon; Fan, Rong

    2014-01-01

    Despite the presence of the blood-brain barrier (BBB) that restricts the entry of immune cells and mediators into the central nervous system (CNS), a small number of peripheral leukocytes can traverse BBB and infiltrate into the CNS. Cerebrospinal fluid (CSF) is one of the major routes through which trafficking leukocytes migrate into the CNS. Therefore, the number of leukocytes and their phenotypic compositions in CSF may represent important sources to investigate immune-to-brain interaction, or diagnose and monitor neurodegenerative diseases. Due to the paucity of trafficking leucocytes in CSF, a technology capable of efficient isolation, enumeration, and molecular typing of these cells in the clinical settings has not been achieved. In this study, we report on a biofunctionalized silicon nanowire array chip for highly efficient capture and multiplexed phenotyping of rare trafficking leukocytes in small quantities (50 microliters) of clinical CSF specimens collected from neurodegenerative disease patients. The antibody-coated 3D nanostructured materials exhibited vastly improved rare cell capture efficiency due to high-affinity binding and enhanced cell-substrate interactions. Moreover, our platform creates multiple cell capture interfaces, each of which can selectively isolate specific leukocyte phenotype. Comparison with the traditional immunophenotyping using flow cytometry demonstrated that our novel silicon nanowire-based rare cell analysis platform can perform rapid detection and simultaneous molecular characterization of heterogeneous immune cells. Multiplexed molecular typing of rare leukocytes in CSF samples collected from Alzheimer’s disease patients revealed the elevation of white blood cell counts and significant alterations in the distribution of major leukocyte phenotypes. Our technology represents a practical tool potentially for diagnosing and monitoring the pathogenesis of neurodegenerative diseases by allowing an effective hematological analysis of CSF from patients. PMID:24705924

  8. Stochastic model of leukocyte chemosensory movement

    Microsoft Academic Search

    R. T. Tranquillo; D. A. Lauffenburger

    1987-01-01

    We propose a hypothesis for a unified understanding of the persistent and biased random walk behavior of leukocytes exhibiting random motility and chemotaxis, respectively. This hypothesis is based on a description of the leukocyte as an integrated system sensing and responding to a “noisy” receptor signal: random fluctuations inherent in receptor-sensing of chemoattractant concentrations underlie the random walk behavior. Noise

  9. Interstitial leukocyte migration and immune function

    Microsoft Academic Search

    Bettina Weigelin; Peter Friedl

    2008-01-01

    The trafficking of leukocytes into and within lymphoid and peripheral tissues is central to immune cell development, immunosurveillance and effector function. Interstitial leukocyte trafficking is the result of amoeboid polarization and migration, guided by soluble or tissue-bound chemoattractant signals for positioning and local arrest. In contrast to other migration modes, amoeboid movement is particularly suited for scanning cellular networks and

  10. Mammary Leukocyte Response to Drug Therapy

    Microsoft Academic Search

    S. C. Nickerson; M. J. Paape; R. J. Harmon; G. Ziv

    1986-01-01

    The possibility exists that antibiotics and anti-inflammatory agents will be used indiscriminately in attempts to reduce leukocyte or somatic cell counts in mammary secretions to conform with Interstate Milk Shippers quality standards for raw milk to be implemented July 1, 1986. Recent in vivo studies evaluating the effect of intramammary drug injection on milk leukocytes confirmed previous in vitro investigations

  11. Role of the cytoskeleton during leukocyte responses

    Microsoft Academic Search

    Miguel Vicente-Manzanares; Francisco Sánchez-Madrid

    2004-01-01

    The cytoskeleton is a cellular network of structural, adaptor and signalling molecules that regulates most cellular functions that are related to the immune response, including migration, extravasation, antigen recognition, activation and phagocytosis by different subsets of leukocytes. Recently, a large number of regulatory elements and structural constituents of the leukocyte cytoskeleton have been identified. In this review, we discuss the

  12. Role of polymorphonuclear neutrophils in atherosclerosis: Current state and future perspectives

    Microsoft Academic Search

    Roberta Baetta; Alberto Corsini

    2010-01-01

    Contrary to the long-standing and widely accepted belief that polymorphonuclear neutrophils (PMN) are of marginal relevance in atherosclerosis, evidence revealing a previously unappreciated role of PMN in the process of atherosclerosis is being accumulating. Systemic inflammation involving activated PMN is clearly associated with unstable conditions of coronary artery disease and an increased number of circulating neutrophils is a well-known risk

  13. Optimizing Viable Leukocyte Sampling from the Female Genital Tract for Clinical Trials: An International Multi-Site Study

    PubMed Central

    De Rosa, Stephen C.; Martinson, Jeffrey A.; Plants, Jill; Brady, Kirsten E.; Gumbi, Pamela P.; Adams, Devin J.; Vojtech, Lucia; Galloway, Christine G.; Fialkow, Michael; Lentz, Gretchen; Gao, Dayong; Shu, Zhiquan; Nyanga, Billy; Izulla, Preston; Kimani, Joshua; Kimwaki, Steve; Bere, Alfred; Moodie, Zoe; Landay, Alan L.; Passmore, Jo-Ann S.; Kaul, Rupert; Novak, Richard M.; McElrath, M. Juliana; Hladik, Florian

    2014-01-01

    Background Functional analysis of mononuclear leukocytes in the female genital mucosa is essential for understanding the immunologic effects of HIV vaccines and microbicides at the site of HIV exposure. However, the best female genital tract sampling technique is unclear. Methods and Findings We enrolled women from four sites in Africa and the US to compare three genital leukocyte sampling methods: cervicovaginal lavages (CVL), endocervical cytobrushes, and ectocervical biopsies. Absolute yields of mononuclear leukocyte subpopulations were determined by flow cytometric bead-based cell counting. Of the non-invasive sampling types, two combined sequential cytobrushes yielded significantly more viable mononuclear leukocytes than a CVL (p<0.0001). In a subsequent comparison, two cytobrushes yielded as many leukocytes (?10,000) as one biopsy, with macrophages/monocytes being more prominent in cytobrushes and T lymphocytes in biopsies. Sample yields were consistent between sites. In a subgroup analysis, we observed significant reproducibility between replicate same-day biopsies (r?=?0.89, p?=?0.0123). Visible red blood cells in cytobrushes increased leukocyte yields more than three-fold (p?=?0.0078), but did not change their subpopulation profile, indicating that these leukocytes were still largely derived from the mucosa and not peripheral blood. We also confirmed that many CD4+ T cells in the female genital tract express the ?4?7 integrin, an HIV envelope-binding mucosal homing receptor. Conclusions CVL sampling recovered the lowest number of viable mononuclear leukocytes. Two cervical cytobrushes yielded comparable total numbers of viable leukocytes to one biopsy, but cytobrushes and biopsies were biased toward macrophages and T lymphocytes, respectively. Our study also established the feasibility of obtaining consistent flow cytometric analyses of isolated genital cells from four study sites in the US and Africa. These data represent an important step towards implementing mucosal cell sampling in international clinical trials of HIV prevention. PMID:24454917

  14. [The noninvasive technique of analysis of aggregation activity of thrombocytes, leukocytes and erythrocytes].

    PubMed

    Kuznik, B I; Fa?n, I A; Kaminski?, A V; Maksimova, O G; Kustovskaia, E M; Bogdanova, Iu V; Zhdanovich, N G; Rodnina, O S; Khasanova, N V

    2013-04-01

    The article considers the noninvasive technique of analysis of aggregation activity of thrombocytes and formation of leukocytic-erythrocytic-thrombocytic aggregates and some particular indicators of hemostasis system. The speckle-analysis of characteristics of coherent light dispersion from surface of erythrocytes moving in artificially isolated vessel section. The study established high correlation ratio between light dispersion index and spontaneous ADP-, adrenalin- and collagen-induced aggregation of thrombocytes, formation of leukocytic-erythrocytic and thrombocytic-erythrocytic aggregations. The established facts permits to consider that analysis noninvasive technique makes it possible to estimate approximately the aggregation activity of blood corpuscles. PMID:23984549

  15. [The phagocytosis of polymorphonuclear neutrophilic granulocytes in progressive periodontitis].

    PubMed

    Konopka, T; Zietek, M

    1995-01-01

    The aim of this paper was the evaluation of the phagocytic activity of neutrophils in blood and in gingival pocket fluid in patients suffering from rapidly progressive periodontitis (RPP) and postjuvenile periodontitis (PJP). Prior to periodontal treatment the authors evaluated the capacity to phagocytose latex particles of peripheral blood neutrophils from 21 patients with RPP, 51 with PJP and 59 healthy subjects (control group) as well as the phagocytic activity of neutrophils in pocket fluid from 21 patients with RPP, 14 with PJP and from 20 healthy subjects. This phagocytic activity was significantly lower in all examined groups in comparison with the control group. A similar evaluation executed 3 months after treatment revealed normal phagocytosis of blood neutrophils from patients with RPP. In patients receiving complementary pharmacotherapy (spiramycine combined with metronidazol), a better improvement of phagocytosis was noted, than that observed in patients treated only surgically. PMID:7481699

  16. Intranuclear crystalloids associated with abnormal granules in eosinophilic leukocytes

    SciTech Connect

    Parmley, R.T.; Crist, W.M.; Roper, M.; Takagi, M.; Austin, R.L.

    1981-12-01

    Ultrastructural evaluation of eosinophilic leukocytes from a 2-yr-old asymptomatic girl with chronic benign neutropenia (CBN) revealed a variety of morphological abnormalities. All eosinophils obtained from blood and marrow specimens contained multiple microcrystalloids in most of the mature cytoplasmic granules. An increase in crystalloid-free, immature granules in late (bilobed nuclei) eosinophils suggested a delay in granule maturation. The eosinophil granules appeared to be of normal size and demonstrated normal acid phosphatase reactivity. Eosinophilic myelocytes contained abnormal cisternae of rough endoplasmic reticulum (RER) and lacked abundant elongated RER cisternae seen in normal cells. A few eosinophilic myelocytes in specimens of bone marrow from the child contained large intranuclear crystalloids measuring up to 3 mu in length. The intranuclear crystalloid contained as cubic lattice of dense material with a periodicity similar to that described for cytoplasmic crystalloids. The ultrastructural morphology of marrow neutrophils was normal, as described in other cases of CBN. Ultrastructural examination of blood eosinophils from the father demonstrated microcrystalloids in cytoplasmic granules identical to those seen in the child. The father was asymptomatic and had normal leukocyte counts. Thus, anomalous crystalloid granule genesis occurred in the father and daughter and was not necessarily associated with neutropenia or clinical symptomatology. This anomaly is associated with the accumulation of intranuclear crystalloid material in eosinophilic myelocytes, which do not appear to be released from the marrow compartment.

  17. Scanning laser ophthalmoscope imaging of fluorescein-labelled blood cells

    Microsoft Academic Search

    Jean-François Le Gargasson; Michel Paques; Jean-Eric Guez; Bernadette Boval; Eric Vicaut; Xin Hou; Yvon Grall; Alain Gaudric

    1997-01-01

    • Purpose: To demonstrate the feasibility of a technique for the visualization by scanning laser ophthalmoscope (SLO) of fluores cein-labelled autologous leukocytes and platelets in retinal vessels. • Method: Individual blood samples from rats and rabbits were centrifuged to isolate platelets and leukocytes, then passively labelled with fluorescein and reinjected into the same animal. An SLO was used to visualize

  18. Morphine inhibits migration of tumor-infiltrating leukocytes and suppresses angiogenesis associated with tumor growth in mice.

    PubMed

    Koodie, Lisa; Yuan, Hongyan; Pumper, Jeffery A; Yu, Haidong; Charboneau, Richard; Ramkrishnan, Sundaram; Roy, Sabita

    2014-04-01

    Tumor cells secrete factors that stimulate the migration of peripheral blood leukocytes and enhance tumor progression by affecting angiogenesis. In these studies, we investigated the effect of morphine, a known immunosuppressant, on leukocyte migration and recruitment to conditioned media derived from long-term cultures of mouse Lewis lung carcinoma cells. Our results indicate that morphine treatment reduced the migration and recruitment of tumor-infiltrating leukocytes into Matrigel plugs and polyvinyl alcohol sponges containing conditioned media derived from long-term cultures of mouse Lewis lung carcinoma cells when compared with placebo. A reciprocal increase in peripheral blood leukocytes was observed at the time of plug or sponge removal in morphine-treated mice. Decreased angiogenesis was observed in conditioned media derived from long-term cultures of mouse Lewis lung carcinoma cells Matrigel plugs taken from morphine-treated wild-type mice when compared with placebo but was abolished in morphine-treated ?-opioid receptor knockout mice. In addition, in vitro studies using trans-well and electric cell substrate impedance sensing system studies reveal for the first time morphine's inhibitory effects on leukocyte migration and their ability to transmigrate across an activated endothelial monolayer. Taken together, these studies indicate that morphine treatment can potentially decrease leukocyte transendothelial migration and reduce angiogenesis associated with tumor growth. The use of morphine for cancer pain management may be beneficial through its effects on angiogenesis. PMID:24495739

  19. Morphine Inhibits Migration of Tumor-Infiltrating Leukocytes and Suppresses Angiogenesis Associated with Tumor Growth in Mice

    PubMed Central

    Koodie, Lisa; Yuan, Hongyan; Pumper, Jeffery A.; Yu, Haidong; Charboneau, Richard; Ramkrishnan, Sundaram; Roy, Sabita

    2015-01-01

    Tumor cells secrete factors that stimulate the migration of peripheral blood leukocytes and enhance tumor progression by affecting angiogenesis. In these studies, we investigated the effect of morphine, a known immunosuppressant, on leukocyte migration and recruitment to conditioned media derived from long-term cultures of mouse Lewis lung carcinoma cells. Our results indicate that morphine treatment reduced the migration and recruitment of tumor-infiltrating leukocytes into Matrigel plugs and polyvinyl alcohol sponges containing conditioned media derived from long-term cultures of mouse Lewis lung carcinoma cells when compared with placebo. A reciprocal increase in peripheral blood leukocytes was observed at the time of plug or sponge removal in morphine-treated mice. Decreased angiogenesis was observed in conditioned media derived from long-term cultures of mouse Lewis lung carcinoma cells Matrigel plugs taken from morphine-treated wild-type mice when compared with placebo but was abolished in morphine-treated ?-opioid receptor knockout mice. In addition, in vitro studies using trans-well and electric cell substrate impedance sensing system studies reveal for the first time morphine's inhibitory effects on leukocyte migration and their ability to transmigrate across an activated endothelial monolayer. Taken together, these studies indicate that morphine treatment can potentially decrease leukocyte transendothelial migration and reduce angiogenesis associated with tumor growth. The use of morphine for cancer pain management may be beneficial through its effects on angiogenesis. PMID:24495739

  20. Weaning management of newly received beef calves with or without continuous exposure to a persistently infected bovine viral diarrhea virus pen mate: effects on health, performance, bovine viral diarrhea virus titers, and peripheral blood leukocytes.

    PubMed

    Richeson, J T; Kegley, E B; Powell, J G; Beck, P A; Ley, B L Vander; Ridpath, J F

    2012-06-01

    Exposure to animals persistently infected (PI) with bovine viral diarrhea virus (BVDV) results in immunomodulation of cohorts that may have health and growth consequences; however, effects may differ in low-risk, preconditioned (PC) vs. high-risk, auction market (AM) beef cattle. Our objective was to compare health and performance of PC or AM management systems with (PI) or without (CON) presence of a PI-BVDV pen mate using a 2 × 2 factorial arrangement. Four shipment blocks of crossbred PC steers (n = 236) from 3 ranch-origins were weaned, dewormed, vaccinated, tested for PI-BVDV, and kept on the ranch for ?42 d. Subsequently, PC steers were transported to a stocker receiving unit (RU), weighed (251 ± 2 kg), blood sampled, stratified by d -1 BW, and assigned randomly to treatment (PCPI or PCCON) with no additional processing. Simultaneously, 4 blocks of crossbred AM calves (n = 292) were assembled from regional auction markets and transported to the RU ± 36 h from PC arrival. The AM calves were weighed (245 ± 1.3 kg), stratified by gender and d -1 BW, processed under the same regimen used for PC steers at their origin ranch except bull calves were castrated, and then assigned randomly to treatment (AMPI or AMCON). Treatment pens (0.45 ha) were arranged spatially such that PI did not have fence-line or water source contact with CON. Calves were fed identically and followed the same antibiotic treatment protocol. Daily BW gain for the entire 42-d receiving trial was greater (P < 0.001) for PC (1.2 kg) compared with AM (0.85 kg). There was an exposure effect (P = 0.002) on ADG from d 28 to 42; CON gained 1.12 kg vs. 0.90 kg BW for PI cohort. Morbidity was markedly greater (P < 0.001) in AM (70%) vs. PC (7%), resulting in (P < 0.001) an antibiotic treatment cost of $20.52 and $2.48/animal, respectively. Treatment with a third antibiotic occurred more often (P = 0.04) for PI cohort, and the percentage of chronically ill cattle was greatest (P = 0.06) for AMPI. Upon arrival, BVDV type 1a, 1b, and 2a titers were greater for PC (treatment × day, P < 0.001), and the percentage seropositive to BVDV type 1a on d 0 was 100% for PC vs. 23% in AM. Platelets increased transiently (P < 0.001) with greater platelets observed in AM (P < 0.001). Results indicate that PC calves gain faster and require fewer antibiotic treatments during the receiving period. Exposure to PI reduced BW gain from d 28 to 42, increased the number of calves treated thrice, and increased chronically ill cattle for AM. PMID:22648754

  1. Leukocyte Telomere Length and Coronary Artery Calcium.

    PubMed

    Hunt, Steven C; Kimura, Masayuki; Hopkins, Paul N; Carr, J Jeffrey; Heiss, Gerardo; Province, Michael A; Aviv, Abraham

    2015-07-15

    Patients with histories of myocardial infarction display shortened leukocyte telomere length (LTL), but conflicting findings have been reported on the relation between LTL and subclinical coronary artery atherosclerosis, as expressed by coronary artery calcium (CAC). The aim of this study was to examine the relation between LTL, measured by Southern blots, and CAC in 3,169 participants in the National Heart, Lung, and Blood Institute Family Heart Study. Participants consisted of 2,556 whites, 613 blacks, 1,790 women, and 1,379 men. The odds of having CAC ?100 for the shortest LTL tertile versus the longest LTL tertile were 1.95 (95% confidence interval [CI] 1.28 to 3.16) in white men and 1.76 (95% CI 1.18 to 2.45) in white women, after adjusting for multiple covariates of CAC. The corresponding odds ratios for blacks were 1.53 (95% CI 0.67 to 3.50) and 0.87 (95% CI 0.37 to 2.00). Significance levels of tests for trend across LTL tertiles were p = 0.002 in white men, p = 0.006 in white women, p = 0.32 in black men, and p = 0.74 in black women. The associations, or lack of associations, were independent of C-reactive protein levels and other risk factors for CAC. As previously shown in other studies, whites displayed shorter LTLs than blacks (p <0.0001). In conclusion, the higher the coronary artery atherosclerotic burden in whites, the shorter the LTL. This LTL-atherosclerosis connection is not found in blacks. The mechanisms for the racial difference in LTL, CAC, and their interrelations do not seem to be related to inflammation and merit further research. PMID:25960381

  2. Measuring leukocyte-endothelial interactions in mice.

    PubMed

    Jain, Rakesh K; Munn, Lance L; Fukumura, Dai

    2013-06-01

    Intravital imaging techniques are powerful tools in the study of tumor pathophysiology. These techniques can be used to measure vascular parameters (e.g., angiogenesis, hemodynamics, and leukocyte-endothelial interactions) in tumors and normal vascular networks in mice. This protocol describes the use of single-photon microscopy and multiphoton laser-scanning microscopy to follow the migration of Rhodamine-6G, a fluorescently labeled tracer, following injection into the tail vein of a mouse. The Rhodamine-6G enters the circulatory system and labels leukocytes. It is possible to monitor changes in the interactions between leukocytes and the endothelium by determining the numbers of rolling and adhering leukocytes as well as the total flux of these cells. PMID:23734020

  3. Exposure to Sodium Fluoride Produces Signs of Apoptosis in Rat Leukocytes

    PubMed Central

    Gutiérrez-Salinas, José; Morales-González, José A.; Madrigal-Santillán, Eduardo; Esquivel-Soto, Jaime; Esquivel-Chirino, César; González-Rubio, Manuel García-Luna y; Suástegui-Domínguez, Sigrit; Valadez-Vega, Carmen

    2010-01-01

    Fluoride is naturally present in the earth’s crust and can be found in rocks, coal, and clay; thus, it can be found in small quantities in water, air, plants, and animals. Therefore, humans are exposed to fluoride through food, drinking water, and in the air they breathe. Flouride is essential to maintain bone strength and to protect against dental decay, but if it is absorbed too frequently, it can cause tooth decay, osteoporosis, and damage to kidneys, bones, nerves, and muscles. Therefore, the present work was aimed at determining the effect of intake of sodium fluoride (NaF) as an apoptosis inducer in leukocytes of rats treated for eight weeks with 1 or 50 parts per million (ppm) NaF. Expression of p53, bcl-2, and caspade-3 were used as apoptotic and general metabolism indicators of leukocyte-like indicators of the (INT) oxidation system. Male rats were exposed to NaF (1 and 500 ppm) for eight weeks, and then sacrificed weekly to obtain blood samples. Expression of p53, bcl-2, and caspase-3 were determined in leukocytes by Western blot, and general metabolism of leukocytes was analyzed with a commercial kit. We found changes in the expression of the proteins described, especially when the animals received 50 ppm of NaF. These results indicate that NaF intoxication can be an apoptosis inducer in rat leukocytes treated with the compound for eight weeks. PMID:20957113

  4. Changes in salivary peroxidase and polymorphonuclear neutrophil leucocyte enzyme activities during the menstrual cycle.

    PubMed

    Cockle, S M; Harkness, R A

    1978-10-01

    An elevation in salivary peroxidase activity has been found about the time of ovulation in 14 menstrual cycles in a total of six women. This peak coincided with the ovulatory luteinizing hormone (LH) and oestrogen peak in the four cycles in which endocrine studies were performed. Rises in some polymorphonuclear neutrophil leucocyte enzymes were also seen around ovulation. The possible use of changes in salivary peroxidase as a method for detection of ovulation is discussed. PMID:708662

  5. Gamma-melanocyte-stimulating hormone-like immunoreactivity in blood cells of human eosinophilic patients.

    PubMed

    Johansson, O; Virtanen, M; Hilliges, M; Hansson, L O

    1991-01-01

    The immunohistochemical localization of the peptide gamma-melanocyte-stimulating hormone (gamma-MSH) within human polymorphonuclear leucocytes of blood from eosinophilic patients is described. The gamma-MSH immunoreactivity was observed only in neutrophilic granulocytes leaving all other cell types immuno-negative. PMID:1805488

  6. Rapid quantitative assessment of phagocytic activity of Indium-111 labeled leukocytes by chemiluminescence

    SciTech Connect

    Juni, J.E.; Petry, N.; Wahl, R.L.; Geatti, O.

    1985-05-01

    Indium-111 labeled leukocyte imaging is gaining widespread acceptance. A rapid method for assaying changes in leukocyte viability and phagocytic function during the labeling process would facilitate the evaluation of new labeling techniques and testing of labeled cells before pt injection. The authors have conducted preliminary investigations of chemiluminescence in the clinical evaluation of leukocyte labeling. The chemiluminescence assay may be performed in 30 minutes with only 0.1 ml of whole blood. Zymossan is rapidly introduced to the blood or cell suspension resulting in the emission of light which is then counted by photometer. The amount of light given off by the reaction reflects both the phagocytic function of the cells and the ability of activated phagocytes to generate activated oxygen species. They have evaluated the chemiluminescent activity of normal human leukocyte suspensions both before and after labeling with Indium-111 oxine. The chemiluminescence assay provides a rapid means of evaluating granulocyte function. Correlations of this activity with image quality may provides clues for optimization of labeling techniques.

  7. Functions and oxidative stress status of leukocytes in patients with nephrotic syndrome.

    PubMed

    Akyol, Taner; Bulucu, Fatih; Sener, Osman; Yamanel, Levent; Aydin, Ahmet; Inal, Volkan; Bozoglu, Ergun; Demirkaya, Erkan; Eken, Ayse; Musabak, Ugur

    2007-06-01

    This study was conducted to establish the functions and oxidative stress status in leukocytes of adult patients with nephrotic syndrome. Thirty adult patients with nephrotic syndrome and 32 controls were included. Phagocytosis ability, the killing ability of the micro-organism phagosited of polymorphonuclear leukocytes (PMNL) and monocytes, along with oxidative stress parameters of PMNLs were assessed. There was no statistically significant difference in phagocytosis function of PMNLs and monocytes of patients when compared to those of controls. PMNL burst activities of the patient and control groups also showed no difference; however, the monocyte burst activities of patients were significant (p = 0.012). The glutathione peroxidase (GSH-Px) activities in PMNLs of the patients with nephrotic syndrome were significantly higher (p = 0.026) when compared to those of controls. In comparison with those of the control subjects, the patients had also higher selenium levels in their PMNLs (p < 0.001). Although PMNL malonyldialdehyde (MDA) levels of the patients seem to be higher than those of controls, the difference had no statistical significance (p = 0.071). Conclusively, in the patients with nephrotic syndrome, PMNLs appear to be exposed to an oxidative stress as indicated by their increased GSH-Px activities and selenium content. However, PMNLs in nephrotic syndrome patients seem to be coping with the insulting oxidative stress, as suggested by their near-normal MDA productions. Furthermore, these data suggest that nephrotic syndrome appears not to have an influence on phagocytosis and killing abilities of granulocytes and monocytes as long as these cells can overcome the oxidative stress to which they are exposed in this disease. PMID:17709904

  8. The inhibitory effects of antirheumatic drugs on the activity of human leukocyte elastase and cathepsin G.

    PubMed

    Steinmeyer, J; Kalbhen, D A

    1996-07-01

    The serine proteinases elastase and cathepsin G from polymorphonuclear granulocytes play a critical role in articular cartilage degradation, not only as proteolytic enzymes able to degrade the extracellular matrix but also by additionally modulating the level of active matrix metalloproteinases, key enzymes of the proteolytic destruction of cartilage during rheumatoid arthritis. The aim of our study was to examine whether anti-inflammatory drugs and selected compounds inhibited elastase and cathepsin G, and also to determine whether it is necessary to use a highly purified elastase preparation to screen drugs for their ability to block the activity of this enzyme. Eglin C and the glycosaminoglycan-peptide complex DAK-16, at concentrations ranging from 10(-9) to 10(-4) M, dose-dependently inhibited elastase and cathepsin G while the nonsteroidal anti-inflammatory drugs oxyphenbutazone, phenylbutazone, sulfinpyrazone and diclofenac-Na required high concentrations to demonstrate some inhibitory effects on the activity of both enzymes. None of the other anti-inflammatory drugs tested at a concentration of 10(-4) M such as acetylsalicylic acid, dexamethasone, indomethacin, ketoprofen, naproxen, oxaceprol, pirprofen and tiaprofenic acid demonstrated any marked inhibitory activity on either of these proteinases. Only a few drugs, when dosed therapeutically, achieved synovial fluid concentrations sufficient to inhibit the activities of both proteinases. The antirheumatic drugs demonstrated similar inhibition profiles in purified or partially purified elastase preparations. Thus the leukocyte extract containing the partially purified elastase and cathepsin G which can be rapidly and easily prepared at low costs appears to be an efficient mean of screening potentially new therapeutic agents for their ability to inhibit leukocyte elastase and cathepsin G. PMID:8841833

  9. Diagnosing bacterial peritonitis made easy by use of leukocyte esterase dipsticks

    PubMed Central

    Chugh, Kiran; Agrawal, Yuthika; Goyal, Vipin; Khatri, Vinod; Kumar, Pradeep

    2015-01-01

    Introduction: Spontaneous bacterial peritonitis (SBP) requires rapid diagnosis for the initiation of antibiotics. Its diagnosis is usually based on manual examination of ascitic fluid (AF) having long reporting time. AF infection is diagnosed when the fluid polymorphonuclear leukocyte (PMNL) concentration ?250 cells/mm3. Aims and Objectives: Aim was to evaluate the diagnostic utility of leukocyte esterase (LE) reagent strip for rapid diagnosis of SBP in patients who underwent abdominal paracentesis and to calculate the sensitivity, specificity, positive, and negative predictive values. Materials and Methods: The study was carried out on 103 patients with ascites. Cell count of AF as determined by colorimetric scale of Multistix 10 SG reagent strip was compared with counting chamber method (PMNL count ?250 cells/mm3 was considered positive). Results and Observations: Of the 103 patients SBP was diagnosed in 20 patients, 83 patients were negative for SBP by manual cell count. The sensitivity and specificity of the LE test for detecting neutrocytic SBP taking grade 2 as cut off were 95% and 96.4% respectively, with a positive predictive value of 86.4% and a negative predictive value of 98.8%. Diagnostic accuracy of LE test was 96.1%. Discussion: There was a good correlation between the reagent strip result and PMNL count. The LE strip test is based on the esterase activity of activated granulocytes which reacts with an ester-releasing hydroxyphenylpyrrole causing a colour change in the azo dye of reagent strip. It is a very sensitive and specific method for the prompt detection of elevated PMNL count, and represents a convenient, inexpensive, simple, and bedside method for diagnosis of SBP. A negative LE test result excludes SBP with a high degree of certainty. PMID:25810962

  10. The influence of donepezil and EGb 761 on the innate immunity of human leukocytes: effect on the NF-?B system.

    PubMed

    Sochocka, Marta; Zaczy?ska, Ewa; Tabo?, Agnieszka; Czarny, Anna; Leszek, Jerzy; Sobczy?ski, Maciej

    2010-12-01

    Ginkgo biloba special extract EGb 761 and donepezil are drugs used in Alzheimer therapy. The influence of donepezil and EGb 761 on two mechanisms of innate immunity, natural antiviral resistance of human leukocytes ex vivo and NF-?B activation, was studied. Correlation between the innate immunity of leukocytes and NF-?B activation was investigated. The effect of the two drugs on resistance of human leukocytes to vesicular stomatitis virus (VSV) infection was also assessed. Two groups of healthy blood donors (n=30) were distinguished: one with resistant leukocytes (n=15) and one (n=15) with leukocytes sensitive to VSV. The degree of natural resistance of human peripheral blood leukocytes (PBLs) was determined by studying the kinetics of VSV replication. NF-?B activation was assayed by immunocytochemical staining. Efficiency of donepezil and EGb 761 was determined by a special regression model. The toxicity of the preparations to PBLs and the cell lines L(929) and A(549) and their effect on the different viruses was established. Results showed that donepezil used in concentrations of 10-50 ?g/ml and EGb761 of 25-100 ?g/ml stimulated resistance of human leukocytes. At the same concentrations both preparations decreased activation of transcriptional factor NF-?B. Correlation between innate immunity of PBLs and NF-?B activation was observed. Comparison of the effects of these two drugs showed that EGb 761 is more effective in stimulating leukocyte resistance. Donepezil and EGb 761 regulated innate immunity of human leukocytes by stimulating resistance and modulating NF-?B activation. The natural drug was more efficient in stimulating innate antiviral immunity of human leukocytes. PMID:20837172

  11. Physical Activity, Sedentary Behavior, and Leukocyte Telomere Length in Women

    PubMed Central

    Du, Mengmeng; Prescott, Jennifer; Kraft, Peter; Han, Jiali; Giovannucci, Edward; Hankinson, Susan E.; De Vivo, Immaculata

    2012-01-01

    Leukocyte telomere length (LTL) is a potential indicator of cellular aging; however, its relation to physical activity and sedentary behavior is unclear. The authors examined cross-sectionally associations among activity, sedentary behavior, and LTL among 7,813 women aged 43–70 years in the Nurses’ Health Study. Participants self-reported activity by questionnaire in 1988 and 1992 and sedentary behavior in 1992. Telomere length in peripheral blood leukocytes, collected in 1989–1990, was measured by quantitative polymerase chain reaction. The least-squares mean telomere length (z-score) was calculated after adjustment for age and other potential confounders. For total activity, moderately or highly active women had a 0.07-standard deviation (SD) increase in LTL (2-sided Ptrend = 0.02) compared with those least active. Greater moderate- or vigorous-intensity activity was also associated with increased LTL (SD = 0.11 for 2–4 vs. <1 hour/week and 0.04 for ?7 vs. <1 hour/week; 2-sided Ptrend = 0.02). Specifically, calisthenics or aerobics was associated with increased LTL (SD = 0.10 for ?2.5 vs. 0 hours/week; 2-sided Ptrend = 0.04). Associations remained after adjustment for body mass index. Other specific activities and sitting were unassociated with LTL. Although associations were modest, these findings suggest that even moderate amounts of activity may be associated with longer telomeres, warranting further investigation in large prospective studies. PMID:22302075

  12. The rainbow trout (Salmo gairdneri Richardson) granular leukocytes and mast-cells system versus the human one. Cytomorphometrical and cytochemical characters in these two species philogenetically poles apart.

    PubMed

    Passantino, L; Patruno, R; Ranieri, G; Cianciotta, A; Sciscioli, V; Passantino, G F

    2000-01-01

    In man, circulating granular leukocytes constitute a cellular system and are able to migrate in the tissues to take part in the immune reactions. This study was to characterize the granular leukocytes and the eventual existence of mast-cells of a low vertebrate, rainbow trout (Salmo gairdneri R.) in order to compare it with man. The blood smears and tissues sections have been tested whit panoptic methods, cytochemistry and toluidine blue. White blood cell count, leukocytes formula and cytomorphometric characterization was performed using an image analyser. Scanning of tissues sections in order to identify mast-cells has been also performed. In this model the granular leukocytes are all neutrophilic like; no eosinophilic, no basophilic no tissue mast-cells, basically existing in allergic and anaphylactic reactions, were found. PMID:11103851

  13. Inhibitory effect of locally administered heparin on leukocyte rolling and chemoattractant-induced firm adhesion in rat mesenteric venules in vivo

    PubMed Central

    Xie, X; Thorlacius, H; Raud, J; Hedqvist, P; Lindbom, L

    1997-01-01

    Anti-inflammatory actions of heparin and related glycosaminoglycans have been described in the literature. Here, we used intravital microscopy of the rat mesentery microcirculation to examine effects of locally administered heparin on leukocyte rolling and chemoattractant-induced firm adhesion. It was found that topical application of heparin reduced N-formyl-methionyl-leucyl-phenylalanine (fMLP)-induced leukocyte adhesion. Notably, the inhibitory action of heparin was not dose-dependent, but rather a biphasic dose-response was found, i.e. low (2 and 20?iu?ml?1) and high (1000?iu?ml?1) concentrations of heparin significantly reduced adhesion, whereas an intermediate dose (200?iu?ml?1) was less effective. Heparin, 2 and 20?iu?ml?1, decreased rolling leukocyte flux, while having no effect on blood flow or total leukocyte flux. By contrast, heparin, 200 and 1000?iu?ml?1, increased total leukocyte flux in parallel with a rise in volume blood flow resulting in recovery of the rolling leukocyte flux at these doses. Thus, the biphasic inhibitory action of heparin on fMLP-induced firm adhesion could in part be attributed to changes in leukocyte delivery (i.e. blood flow) and rolling leukocyte flux induced by heparin. When compensating for the influence of different rolling levels on fMLP-evoked adhesion, a dose-related inhibitory effect of heparin on the firm adhesive response per se was revealed. Similar results were obtained in a static adhesion assay in vitro where heparin 200 and 1000?iu?ml?1 (but not 2 and 20?iu?ml?1) significantly inhibited fMLP-induced leukocyte adhesion in the absence of any modulatory influence on changes in rolling. Our data show that locally administered heparin inhibits leukocyte rolling as well as chemoattractant-induced firm adhesion in vivo which thus may contribute to the postulated anti-inflammatory activity of this compound. However, because of interference with several microvascular functions, strict dose-dependent responses to heparin treatment were not found, which illustrates the complex interplay between local blood flow, leukocyte rolling and chemoattractant-induced adhesion as determinants of leukocyte recruitment to tissues in inflammation. PMID:9384507

  14. A Leukocyte Score to Improve Clinical Outcome Predictions in Bacteremic Pneumococcal Pneumonia in Adults

    PubMed Central

    Blot, Mathieu; Croisier, Delphine; Péchinot, André; Vagner, Ameline; Putot, Alain; Fillion, Aurélie; Baudouin, Nicolas; Quenot, Jean-Pierre; Charles, Pierre-Emmanuel; Bonniaud, Philippe; Chavanet, Pascal; Piroth, Lionel

    2014-01-01

    Background ?Bacteremic pneumococcal pneumonia (BPP) is associated with high and early mortality. A simple procedure to predict mortality is crucial. Methods ?All adult patients with BPP admitted from 2005 through 2013 to the University Hospital of Dijon, France, were enrolled to study 30-day mortality and associated factors, particularly leukocyte counts. A simple leukocyte score was created by adding 1 point each for neutropenia (<1500 cells/mm3), lymphopenia (<400), and monocytopenia (<200). Results ?One hundred and ninety-two adult patients (mean age, 69 years; standard deviation [SD], 19 years) who had developed and were hospitalized for BPP (58% community-acquired) were included. The 30-day crude mortality rate was 21%. The mean Pneumonia Severity Index score was high at 127.3 (SD = 41.3). Among the 182 patients who had a white blood cell count, 34 (19%) had a high leukocyte score (?2). Multivariate analysis revealed that mortality was significantly associated with a high leukocyte score (odds ratio, 6.28; 95% confidence interval, 2.35–16.78), a high respiratory rate, a low serum bicarbonate level, and an altered mental status (all P < .05). The leukocyte score was not significantly dependent on the previous state of immunosuppression, alcoholism, or viral coinfection, but it did correlate with an acute respiratory distress syndrome and a low serum bicarbonate level. Conclusions ?This new leukocyte score, in combination with the well known predictive factors, seems of interest in predicting the risk of death in BPP. A high score correlated with organ dysfunction and probably reflects the level of immunoparalysis. Its predictive value has to be confirmed in other cohorts. PMID:25734145

  15. Triggering of leukocytes by phase contrast in imaging cytometry with scanning fluorescence microscope (SFM)

    NASA Astrophysics Data System (ADS)

    Bocsi, József; Pierzchalski, Arkadiusz; Marecka, Monika; Malkusch, Wolf; Tárnok, Attila

    2009-02-01

    Slide-based cytometry (SBC) leads to breakthrough in cytometry of cells in tissues, culture and suspension. Carl Zeiss Imaging Solutions' new automated SFM combines imaging with cytometry. A critical step in image analysis is selection of appropriate triggering signal to detect all objects. Without correct target cell definition analysis is hampered. DNA-staining is among the most common triggering signals. However, the majority of DNA-dyes yield massive spillover into other fluorescence channels limiting their application. By microscopy objects of >5?m diameter can be easily detected by phase-contrast signal (PCS) without any staining. Aim was to establish PCS - triggering for cell identification. Axio Imager.Z1 motorized SFM was used (high-resolution digital camera, AxioCam MRm; AxioVision software: automatic multi-channel scanning, analysis). Leukocytes were stained with FITC (CD4, CD8) and APC (CD3) labelled antibodies in combinations using whole blood method. Samples were scanned in three channels (PCS/FITC/APC). Exposition-times for PCS were set as low as possible; the detection efficiency was verified by fluorescence. CD45-stained leukocytes were counted and compared to the number of PCS detected events. Leukocyte subtyping was compared with other cytometers. In focus the PCS of cells showed ring-form that was not optimal for cell definition. Out of focus PCS allows more effective qualitative and quantitative cell analyses. PCS was an accurate triggering signal for leukocytes enabling cell counting and discrimination of leukocytes from platelets. Leukocyte subpopulation frequencies were comparable to those obtained by other cytometers. In conclusion PCS is a suitable trigger-signal not interfering with fluorescence detection.

  16. 3,4-di-deoxyglucosone-3-ene promotes leukocyte apoptosis

    Microsoft Academic Search

    MARINA PENÉLOPE CATALAN; BEATRIZ SANTAMARÍA; ANA REYERO; ARTURO ORTIZ; JESÚS EGIDO; ALBERTO ORTIZ

    2005-01-01

    3,4-di-deoxyglucosone-3-ene promotes leukocyte apoptosis.BackgroundHeat-sterilized, single-chambered, glucose-containing peritoneal dialysis solutions promote neutrophil apoptosis and impair the peritoneal antibacterial response. It has been proposed that glucose degradation products may be responsible for this effect. However, the precise contribution of individual glucose degradation products had not been addressed.MethodsThe effect of individual glucose degradation products on apoptosis in cultured human neutrophils and peripheral blood

  17. Characterization of two novel LPS-binding sites in leukocyte integrin  A domain

    Microsoft Academic Search

    Kwong-Fai Wong; John M. Luk; R. Holland Cheng; Lloyd B. Klickstein; Sheung-Tat Fan

    2007-01-01

    Lipopolysaccharide (LPS), a bacterial endotoxin, triggers deleterious systemic inflammatory responses when released into blood circulation, causing organ dysfunction and death. In response to LPS stimulation, CD14 and toll-like receptor (TLR)-4 elicit inflammatory signaling cascades. Although leukocyte integrins (CD11b\\/CD18 and CD11c\\/CD18) were re- ported to bind LPS and induce NF-B translocation, the evidence on such epitope location remains elusive. The present

  18. Leukocytapheresis therapy, performed with leukocyte removal filter, for inflammatory bowel disease

    Microsoft Academic Search

    Koji Sawada; Kunio Ohnishi; Shin Fukui; Kazuhiko Yamada; Makoto Yamamura; Kuniki Amano; Kanzo Amano; Masaaki Wada; Noritoshi Tanida; Masamichi Satomi; Takashi Shimoyama

    1995-01-01

    Leukocytapheresis (LCAP), performed with a leukocyte removal filter, was administered five times, at 1-week intervals, for\\u000a 5 weeks of intensive therapy and five times, at approximately 1-month intervals, for approximately 5 months of maintenance\\u000a therapy, to 13 patients with inflammatory bowel disease (IBD) diagnosed as ulcerative colitis (UC) in 8 and Crohn's disease\\u000a (CD) in 5. Clinical and blood examinations

  19. Osteomyelitis: diagnosis with In-111-labeled leukocytes

    SciTech Connect

    Schauwecker, D.S.

    1989-04-01

    In a retrospective review, 485 patients with suspected osteomyelitis were studied. Of these, 453 patients were studied with both bone and indium-111 leukocyte scanning (173 sequentially and 280 simultaneously). The ability to determine that the infection was in bone rather than in adjacent soft tissue was greater with simultaneous bone scan and In-111 leukocyte studies than with sequential studies. The locations of suspected osteomyelitis were divided into central (containing active bone marrow), peripheral (hands and feet), and middle (between central and peripheral). Specificity remained high (about 90%) regardless of the location. Overall sensitivity was significantly lower in the central location than in the peripheral or middle location. Determination of whether the In-111 leukocyte activity was in bone or adjacent soft tissue was also more difficult when the infection was in the central location. For acute osteomyelitis, sensitivity was high regardless of the location. For chronic osteomyelitis, sensitivity was lower in the central location.

  20. Intravascular leukocyte migration through platelet thrombi: directing leukocytes to sites of vascular injury.

    PubMed

    Ghasemzadeh, M; Hosseini, E

    2015-05-26

    Leukocytes recruitment to thrombi supports an intimate cellular interaction leading to the enhancement of pro-coagulant functions and pro-inflammatory responses at site of vascular injury. Recent observations of neutrophil extracellular traps (NETs) formation and its mutual reactions with platelet thrombi adds more clinical interest to the growing body of knowledge in the field of platelet-leukocyte cross-talk. However, having considered thrombus as a barrier between leukocytes and injured endothelium, the full inflammatory roles of these cells during thrombosis is still ill defined. The most recent observation of neutrophils migration into the thrombi is a phenomenon that highlights the inflammatory functions of leukocytes at the site of injury. It has been hypothesised that leukocytes migration might be associated with the conveyance of highly reactive pro-inflammatory and/or pro-coagulant mediators to sites of vascular injury. In addition, the evidence of neutrophils migration into arterial thrombi following traumatic and ischaemia-reperfusion injury highlights the already described role of these cells in atherosclerosis. Regardless of the mechanisms behind leukocyte migration, whether these migrated cells benefit normal homeostasis by their involvement in wound healing and vascular rebuilding or they increase unwilling inflammatory responses, could be of interest for future researches that provide new insight into biological importance of leukocyte recruitment to thrombi. PMID:25880990

  1. Effect of Commiphora molmol on leukocytes proliferation in relation to histological alterations before and during healing from injury

    PubMed Central

    Haffor, Al-Said A.

    2010-01-01

    Myrrh, Commiphora molmol has been used as anti-inflammatory and wound healing commercial product. Leukocyte count had been reliable indicator for clinicians to monitor progress of healing for their patients. We hypothesized that myrrh supplement participate in the activation of leukocyte proliferation pathway prior and post skin injury and gastric ulcer. The purpose of the present study was to examine long-term effect of myrrh on leukocytes proliferation before injury and during different stages of healing. Results showed that all types of leukocytes were significantly (p < 0.05) higher in the myrrh-treated groups before and during healing. The pretreatment with myrrh offered a time-dependent rise in leukocytes proliferation. Microscopic examination of blood smear from myrrh-treated rats with skin injury, showed an elevated count of middle-sized lymphocytes and neutrophils that were characterized with well-defined nuclear lobules and rich-granules cytoplasm. Furthermore, the microscopic examinations of the spleen and lymph nodes of myrrh-treated rats with skin injury, showed an increased thickness of lymphatic sheath around the arterioles in the white pulp that was associated with high density of the medium-sized lymphocytes in the secondary lymphoid follicles in the lymph nodes with engorged sinusoids. As myrrh enhanced leukocytes proliferation before injury, it can be concluded that myrrh posse’s antigenic-driven responses and that indicated some foreignness or toxicity of some constituents of myrrh. Because myrrh helped to maintain the relative rise of leukocytes counts throughout healing period and that implied it activated late steps of both proliferation and differentiation pathways for all types of leukocytes during effective phase of the specific immune responses. PMID:23961070

  2. Oxidative stress and reduced responsiveness of challenged circulating leukocytes following pulmonary instillation of metal-rich particulate matter in rats.

    PubMed

    Erdely, Aaron; Antonini, James M; Young, Shih-Houng; Kashon, Michael L; Gu, Ja K; Hulderman, Tracy; Salmen, Rebecca; Meighan, Terence; Roberts, Jenny R; Zeidler-Erdely, Patti C

    2014-01-01

    Welding fume is an exposure that consists of a mixture of metal-rich particulate matter with gases (ozone, carbon monoxide) and/or vapors (VOCs). Data suggests that welders are immune compromised. Given the inability of pulmonary leukocytes to properly respond to a secondary infection in animal models, the question arose whether the dysfunction persisted systemically. Our aim was to evaluate the circulating leukocyte population in terms of cellular activation, presence of oxidative stress, and functionality after a secondary challenge, following welding fume exposure. Rats were intratracheally instilled (ITI) with PBS or 2 mg of welding fume collected from a stainless steel weld. Rats were sacrificed 4 and 24 h post-exposure and whole blood was collected. Whole blood was used for cellular differential counts, RNA isolation with subsequent microarray and Ingenuity Pathway Analysis, and secondary stimulation with LPS utilizing TruCulture technology. In addition, mononuclear cells were isolated 24 h post-exposure to measure oxidative stress by flow cytometry and confocal microscopy. Welding fume exposure had rapid effects on the circulating leukocyte population as identified by relative mRNA expression changes. Instillation of welding fume reduced inflammatory protein production of circulating leukocytes when challenged with the secondary stimulus LPS. The effects were not related to transcription, but were observed in conjunction with oxidative stress. These findings support previous studies of an inadequate pulmonary immune response following a metal-rich exposure and extend those findings showing leukocyte dysfunction occurs systemically. PMID:25123171

  3. Oxidative stress and reduced responsiveness of challenged circulating leukocytes following pulmonary instillation of metal-rich particulate matter in rats

    PubMed Central

    2014-01-01

    Welding fume is an exposure that consists of a mixture of metal-rich particulate matter with gases (ozone, carbon monoxide) and/or vapors (VOCs). Data suggests that welders are immune compromised. Given the inability of pulmonary leukocytes to properly respond to a secondary infection in animal models, the question arose whether the dysfunction persisted systemically. Our aim was to evaluate the circulating leukocyte population in terms of cellular activation, presence of oxidative stress, and functionality after a secondary challenge, following welding fume exposure. Rats were intratracheally instilled (ITI) with PBS or 2 mg of welding fume collected from a stainless steel weld. Rats were sacrificed 4 and 24 h post-exposure and whole blood was collected. Whole blood was used for cellular differential counts, RNA isolation with subsequent microarray and Ingenuity Pathway Analysis, and secondary stimulation with LPS utilizing TruCulture technology. In addition, mononuclear cells were isolated 24 h post-exposure to measure oxidative stress by flow cytometry and confocal microscopy. Welding fume exposure had rapid effects on the circulating leukocyte population as identified by relative mRNA expression changes. Instillation of welding fume reduced inflammatory protein production of circulating leukocytes when challenged with the secondary stimulus LPS. The effects were not related to transcription, but were observed in conjunction with oxidative stress. These findings support previous studies of an inadequate pulmonary immune response following a metal-rich exposure and extend those findings showing leukocyte dysfunction occurs systemically. PMID:25123171

  4. Modeling Leukocyte-Leukocyte Non-Contact Interactions in a Lymph Node

    PubMed Central

    Gritti, Nicola; Caccia, Michele; Sironi, Laura; Collini, Maddalena; D'Alfonso, Laura; Granucci, Francesca; Zanoni, Ivan; Chirico, Giuseppe

    2013-01-01

    The interaction among leukocytes is at the basis of the innate and adaptive immune-response and it is largely ascribed to direct cell-cell contacts. However, the exchange of a number of chemical stimuli (chemokines) allows also non-contact interaction during the immunological response. We want here to evaluate the extent of the effect of the non-contact interactions on the observed leukocyte-leukocyte kinematics and their interaction duration. To this aim we adopt a simplified mean field description inspired by the Keller-Segel chemotaxis model, of which we report an analytical solution suited for slowly varying sources of chemokines. Since our focus is on the non-contact interactions, leukocyte-leukocyte contact interactions are simulated only by means of a space dependent friction coefficient of the cells. The analytical solution of the Keller-Segel model is then taken as the basis of numerical simulations of interactions between leukocytes and their duration. The mean field interaction force that we derive has a time-space separable form and depends on the chemotaxis sensitivity parameter as well as on the chemokines diffusion coefficient and their degradation rate. All these parameters affect the distribution of the interaction durations. We draw a successful qualitative comparison between simulated data and sets of experimental data for DC-NK cells interaction duration and other kinematic parameters. Remarkably, the predicted percentage of the leukocyte-leukocyte interactions falls in the experimental range and depends (?25% increase) upon the chemotactic parameter indicating a non-negligible direct effect of the non-contact interaction on the leukocyte interactions. PMID:24204669

  5. Boundary-precise segmentation of nucleus and plasma of leukocytes

    NASA Astrophysics Data System (ADS)

    Zerfaß, Thorsten; Rehn, Thomas; Wittenberg, Thomas

    2008-03-01

    The exact segmentation of nucleus and plasma of a white blood cell (leukocyte) is the basis for the creation of an automatic, image based differential white blood cell count(WBC). In this contribution we present an approach for the according segmentation of leukocytes. For a valid classification of the different cell classes, a precise segmentation is essential. Especially concerning immature cells, which can be distinguished from their mature counterparts only by small differences in some features, a segmentation of nucleus and plasma has to be as precise as possible, to extract those differences. Also the problems with adjacent erythrocyte cells and the usage of a LED illumination are considered. The presented approach can be separated into several steps. After preprocessing by a Kuwahara-filter, the cell is localized by a simple thresholding operation, afterwards a fast-marching method for the localization of a rough cell boundary is defined. To retrieve the cell area a shortest-path-algorithm is applied next. The cell boundary found by the fast-marching approach is finally enhanced by a post-processing step. The concluding segmentation of the cell nucleus is done by a threshold operation. An evaluation of the presented method was done on a representative sample set of 80 images recorded with LED illumination and a 63-fold magnification dry objective. The automatically segmented cell images are compared to a manual segmentation of the same dataset using the Dice-coefficient as well as Hausdorff-distance. The results show that our approach is able to handle the different cell classes and that it improves the segmentation quality significantly.

  6. Misleading findings on indium-111 leukocyte images

    SciTech Connect

    Goodgold, H.M.; Samuels, L.D.

    1986-06-01

    In-111 leukocyte imaging usually yields images which are interpreted easily. Five potentially misleading cases were noted, however, of over 200 scans performed over a four-year period. These included two cases of bladder visualization without bladder inflammation and three cases of abdominal activity without intra-abdominal abscess or inflammatory bowel disease. Migration of leukocytes from other true pathologic sites and technical factors were responsible for the activity seen. Knowledge of all aspects of the patient's clinical status, including other diagnostic modalities performed, was critical in arriving at the correct diagnosis.

  7. Halo sign on indium-111 leukocyte scan in gangrenous cholecystitis

    SciTech Connect

    Bauman, J.M.; Boykin, M.; Hartshorne, M.F.; Cawthon, M.A.; Landry, A.J.

    1986-02-01

    A 56-year-old man with a long history of Crohn's disease was evaluated by In-111 labeled leukocyte scanning. A halo of leukocyte activity was seen around the gallbladder fossa. A gangrenous gallbladder was removed at surgery.

  8. Lipids on the move: phosphoinositide 3-kinases in leukocyte function

    Microsoft Academic Search

    Matthias P Wymann; Silvano Sozzani; Fiorella Altruda; Alberto Mantovani; Emilio Hirsch

    2000-01-01

    All four known cell surface receptor-activated phosphoinositide 3-kinases have been found in leukocytes. Matthias Wymann and colleagues discuss their nonredundant roles in leukocyte function, particularly in chemotaxis, and their promise as targets for therapeutic intervention.

  9. BRIEF COMMUNICATION Automatic Tracking of Rolling Leukocytes in Vivo

    E-print Network

    Acton, Scott

    , and clutter. Correlation track- ers use the correlation of a fixed template with the image to find instancesBRIEF COMMUNICATION Automatic Tracking of Rolling Leukocytes in Vivo Scott T. Acton,* Klaus Wethmar, the leukocyte position must be tracked, typically by manual observation. Automated tracking of leukocytes

  10. Platelets are essential for leukocyte recruitment in allergic inflammation

    Microsoft Academic Search

    Simon C. Pitchford; Hiroshi Yano; Rebecca Lever; Yanira Riffo-Vasquez; Silvia Ciferri; Mark J. Rose; Silvia Giannini; Stefania Momi; Domenico Spina; Brian O'Connor; Paolo Gresele; Clive P. Page

    2003-01-01

    Background: The role of platelets in inflammation is recognized but poorly characterized, and little is known of their interaction with leukocytes. However, platelet-leukocyte interactions have been demonstrated in cardiovascular disease, culminating in enhanced leukocyte recruitment. Objectives: This study was undertaken to assess the possibility and potential role of similar phenomena occurring in asthmatic patients, a murine model of allergic inflammation,

  11. Leukocyte activation in atherosclerosis: correlation with risk factors

    Microsoft Academic Search

    A. M Elneihoum; P Falke; B Hedblad; F Lindgärde; K Ohlsson

    1997-01-01

    Leukocytes have been implicated in the development of atherosclerotic vascular diseases, and numerous abnormalities of leukocytes in conjunction with atherosclerosis have been reported. The aim of this study of middle-aged asymptomatic subjects with early atherosclerosis was to determine whether a relationship exists between the levels of plasma markers of leukocyte activation, i.e. cytokines and proteases and risk factors for atherosclerosis

  12. Donor leukocyte infusions for multiple myeloma

    Microsoft Academic Search

    M Salama; T Nevill; D Marcellus; P Parker; M Johnson; A Kirk; D Porter; S Giralt; JE Levine; W Drobyski; AJ Barrett; M Horowitz; RH Collins

    2000-01-01

    Donor leukocyte infusion (dli) has well-documented activity in cml, but the role of dli in other diseases is less well defined. to evaluate the strategy in multiple myeloma (mm) we evaluated 25 mm patients from 15 centers who were treated with dli. patients with persistent or recurrent disease after allogeneic bmt received dli from the original marrow donor (23 matched

  13. P2X1 expressed on polymorphonuclear neutrophils and platelets is required for thrombosis in mice

    PubMed Central

    Darbousset, Roxane; Delierneux, Céline; Mezouar, Soraya; Hego, Alexandre; Lecut, Christelle; Guillaumat, Isabelle; Riederer, Markus A.; Evans, Richard J.; Dignat-George, Françoise; Panicot-Dubois, Laurence; Oury, Cécile

    2014-01-01

    Adenosine triphosphate (ATP) and its metabolite, adenosine, are key regulators of polymorphonuclear neutrophil (PMN) functions. PMNs have recently been implicated in the initiation of thrombosis. We investigated the role of ATP and adenosine in PMN activation and recruitment at the site of endothelial injury. Following binding to the injured vessel wall, PMNs are activated and release elastase. The recruitment of PMNs and the subsequent fibrin generation and thrombus formation are strongly affected in mice deficient in the P2X1-ATP receptor and in wild-type (WT) mice treated with CGS 21680, an agonist of the A2A adenosine receptor or NF449, a P2X1 antagonist. Infusion of WT PMNs into P2X1-deficient mice increases fibrin generation but not thrombus formation. Restoration of thrombosis requires infusion of both platelets and PMNs from WT mice. In vitro, ATP activates PMNs, whereas CGS 21680 prevents their binding to activated endothelial cells. These data indicate that adenosine triphosphate (ATP) contributes to polymorphonuclear neutrophil (PMN) activation leading to their adhesion at the site of laser-induced endothelial injury, a necessary step leading to the generation of fibrin, and subsequent platelet-dependent thrombus formation. Altogether, our study identifies previously unknown mechanisms by which ATP and adenosine are key molecules involved in thrombosis by regulating the activation state of PMNs. PMID:25150292

  14. Increased Adenosine Triphosphatase in Leukocytes of Asthmatic Children

    PubMed Central

    Coffey, Ronald G.; Hadden, John W.; Middleton, Elliott

    1974-01-01

    Adenosine triphosphatase (ATPase) activities were compared in leukocytes of asthmatic and nonasthmatic children. Both Mg2+- and Ca2+-dependent ATPase activities were significantly elevated in two membrane fractions (59 to 66%) and in a superntant fraction (68 to 72%) prepared from sonicated leukocytes of asthmatic subjects. Intact cell surface or ecto ATPase was also elevated (67 to 76%) in asthmatic leukocytes. Alternate day glucocorticosteroid therapy was associated with leukocyte ATPase activities intermediate between those for asthmatics not receiving steroids and for nonasthmatic control subjects. Incubation of normal leukocytes with 10-8 M hydrocortisone or leukocyte membranes with 10-4-10-3 M hydrocortisone in vitro also resulted in decreased ATPase activities. The elevated leukocyte ATPase activities appear to relate to the adrenergic imbalance in asthma previously characterized by reduced beta adrenergic responsiveness of adenylate cyclase and suggest the possibility of more than one enzymatic abnormality intrinsic to the asthmatic condition. PMID:4276134

  15. Dielectric properties of human leukocyte subpopulations determined by electrorotation as a cell separation criterion.

    PubMed

    Yang, J; Huang, Y; Wang, X; Wang, X B; Becker, F F; Gascoyne, P R

    1999-06-01

    The separation and purification of human blood cell subpopulations is an essential step in many biomedical applications. New dielectrophoretic fractionation methods have great potential for cell discrimination and manipulation, both for microscale diagnostic applications and for much larger scale clinical problems. To discover whether human leukocyte subpopulations might be separable by such methods, the dielectric characteristics of the four main leukocyte subpopulations, namely, B- and T-lymphocytes, monocytes, and granulocytes, were measured by electrorotation over the frequency range 1 kHz to 120 MHz. The subpopulations were derived from human peripheral blood by magnetically activated cell sorting (MACS) and sheep erythrocyte rosetting methods, and the quality of cell fractions was checked by flow cytometry. Mean specific membrane capacitance values were calculated from the electrorotation data as 10.5 (+/- 3.1), 12.6 (+/- 3.5), 15.3 (+/- 4.3), and 11.0 (+/- 3.2) mF/m2 for T- and B-lymphocytes, monocytes, and granulocytes, respectively, according to a single-shell dielectric model. In agreement with earlier findings, these values correlated with the richness of the surface morphologies of the different cell types, as revealed by scanning electron microscopy (SEM). The data reveal that dielectrophoretic cell sorters should have the ability to discriminate between, and to separate, leukocyte subpopulations under appropriate conditions. PMID:10354456

  16. Candida albicans stimulates cytokine production and leukocyte adhesion molecule expression by endothelial cells.

    PubMed Central

    Filler, S G; Pfunder, A S; Spellberg, B J; Spellberg, J P; Edwards, J E

    1996-01-01

    Endothelial cells have the potential to influence significantly the host immune response to blood-borne microbial pathogens, such as Candida albicans. We investigated the ability (of this organism to stimulate endothelial cell responses relevant to host defense in vitro. Infection with C. albicans induced endothelial cells to express mRNAs encoding E-selectin, intercellular adhesion molecule 1, vascular cell adhesion molecule 1, interleukin 6, interleukin 8, monocyte chemoattractant protein 1, and inducible cyclooxygenase (cox2). All three leukocyte adhesion molecule proteins were expressed on the surfaces of the endothelial cells after 8 h of exposure to C. albicans. An increase in secretion of all three cytokines was found after 12 h of infection. Cytochalasin D inhibited accumulation of the endothelial cell cytokine and leukocyte adhesion molecule mRNAs in response to C. albicans, suggesting that endothelial cell phagocytosis of the organism is required to induce this response. Live Candida tropicalis, Candida glabrata, a nongerminating strain of C. albicans, and killed C. albicans did not stimulate the expression of any of the cytokine or leukocyte adhesion molecule mRNAs. These findings indicate that a factor associated with live, germinating C. albicans is required for induction of endothelial cell mRNA expression. Furthermore, since endothelial cells phagocytize killed C. albicans, phagocytosis is likely necessary but not sufficient for this organism to stimulate mRNA accumulation. In conclusion, the secretion of proinflammatory cytokines and expression of leukocyte adhesion molecules by endothelial cells in response to C. albicans could enhance the host defense against this organism by contributing to the recruitment of activated leukocytes to sites of intravascular infection. PMID:8698486

  17. CXCL1-Triggered Interaction of LFA1 and ICAM1 Control Glucose-Induced Leukocyte Recruitment during Inflammation In Vivo

    PubMed Central

    Buschmann, Kirsten; Koch, Lutz; Braach, Natascha; Mueller, Hanna; Frommhold, David; Poeschl, Johannes; Ruef, Peter

    2012-01-01

    It is well acknowledged that proinflammatory stimulation during acute hyperglycemia is able to aggravate inflammatory diseases. However, the mechanisms of proinflammatory effects of glucose are controversially discussed. We investigated leukocyte recruitment after intravenous injection of glucose in different inflammatory models using intravital microscopy. Flow chamber experiments, expression analysis, functional depletion, and knockout of key adhesion molecules gave mechanistic insight in involved pathways. We demonstrated that a single injection of glucose rapidly increased blood glucose levels in a dose-dependent manner. Notably, during tumor necrosis factor (TNF) ?-induced inflammation leukocyte recruitment was not further enhanced by glucose administration, whereas glucose injection profoundly augmented leukocyte adhesion and transmigration into inflamed tissue in the trauma model, indicating that proinflammatory properties of glucose are stimulus dependent. Experiments with functional or genetic inhibition of the chemokine receptor CXCR2, intercellular adhesion molecule 1 (ICAM1), and lymphocyte function antigen 1 (LFA1) suggest that keratino-derived-chemokine CXCL1-triggered interactions of ICAM1 and LFA1 are crucially involved in the trauma model of inflammation. The lacking effect of glucose on ?2 integrin expression and on leukocyte adhesion in dynamic flow chamber experiments argues against leukocyte-driven underlying mechanisms and favours an endothelial pathway since endothelial ICAM1 expression was significantly upregulated in response to glucose. PMID:23093821

  18. Acute loss of renal function abolishes slow leukocyte rolling and transmigration by interfering with intracellular-signaling molecules

    PubMed Central

    Rossaint, Jan; Spelten, Oliver; Kässens, Nadja; Mueller, Helena; Van Aken, Hugo; Singbartl, Kai; Zarbock, Alexander

    2011-01-01

    Acute kidney injury (AKI) is a common clinical problem in critically ill patients and increases in-hospital mortality. Acute loss of renal function (ALRF) reduces leukocyte recruitment into inflamed tissue, but the underlying molecular mechanisms remain unknown. In this study, we investigated the effects of ALRF on the different steps of the leukocyte recruitment cascade by using intravital microscopy, flow chamber experiments, and biochemistry assays. ALRF abolished selectin-induced slow leukocyte rolling on E-selectin/ICAM-1 and P-selectin/ICAM-1 and also reduced transmigration without affecting chemokine-induced arrest. A reduced phosphorylation of spleen tyrosine kinase (Syk), Akt, phospholipase C (PLC) ?2, and p38 MAPK, but not altered expression levels of adhesion molecules on the surface of neutrophils, was responsible for the abolished selectin-mediated slow leukocyte rolling. The results observed in the murine system could be reproduced in flow chamber experiments with human blood. Samples from critically ill patients with sepsis-induced AKI showed a significantly higher rolling velocity on E-selectin/ICAM-1 and P-selectin/ICAM-1 compared to patients with sepsis alone or to healthy volunteers. In conclusion, these data suggest that ALRF inhibits selectin-mediated slow leukocyte rolling by reducing phosphorylation of Syk, Akt, PLC?2, and p38 MAPK and transmigration. PMID:21562471

  19. [Non-invasive methods for the assessment of platelet, leukocyte, erythrocyte aggregation and coagulation hemostasis].

    PubMed

    Kuznik, B I; Fa?n, I A; Kaminski?, A V; Maksimova, O G; Kustovskaia, E M; Martynova, E N; Rodnina, O S; Khasanova, N V

    2013-01-01

    A non-invasive method for the study of platelet aggregation and formation of leukocyte-erythrocyte-platelet aggregates as well as certain hemostatic parameters is proposed. The method is based on the speckle-analysis of coherent light scattering from the surface of erythrocytes moving in an artificially isolated vessel segment. It was shown that light scattering index significantly correlated with ADP-, adrenalin-, or collagen-induced platelet aggregation, with the formation of leukocyte-erythrocyte or platelet-erythrocyte aggregates, and with the levels of fibrinogen, soluble fibrin-monomer complexes and related parameters. It is concluded that the proposed method for the study of hemostatic system can be used to roughly evaluate intensity of intravascular blood coagulation and probability of thrombosis. PMID:24437188

  20. Synthetic nanoparticles functionalized with biomimetic leukocyte membranes possess cell-like functions

    NASA Astrophysics Data System (ADS)

    Parodi, Alessandro; Quattrocchi, Nicoletta; van de Ven, Anne L.; Chiappini, Ciro; Evangelopoulos, Michael; Martinez, Jonathan O.; Brown, Brandon S.; Khaled, Sm Z.; Yazdi, Iman K.; Enzo, Maria Vittoria; Isenhart, Lucas; Ferrari, Mauro; Tasciotti, Ennio

    2013-01-01

    The therapeutic efficacy of systemic drug-delivery vehicles depends on their ability to evade the immune system, cross the biological barriers of the body and localize at target tissues. White blood cells of the immune system--known as leukocytes--possess all of these properties and exert their targeting ability through cellular membrane interactions. Here, we show that nanoporous silicon particles can successfully perform all these actions when they are coated with cellular membranes purified from leukocytes. These hybrid particles, called leukolike vectors, can avoid being cleared by the immune system. Furthermore, they can communicate with endothelial cells through receptor-ligand interactions, and transport and release a payload across an inflamed reconstructed endothelium. Moreover, leukolike vectors retained their functions when injected in vivo, showing enhanced circulation time and improved accumulation in a tumour.

  1. Indium-111 leukocyte scanning and fracture healing

    SciTech Connect

    Mead, L.P.; Scott, A.C.; Bondurant, F.J.; Browner, B.D. (Univ. of Texas Medical School, Houston (USA))

    1990-01-01

    This study was undertaken to determine the specificity of indium-111 leukocyte scans for osteomyelitis when fractures are present. Midshaft tibial osteotomies were performed in 14 New Zealand white rabbits, seven of which were infected postoperatively with Staphylococcus aureus per Norden's protocol. All 14 rabbits were scanned following injection with 75 microCi of indium 111 at 72 h after osteotomy and at weekly intervals for 4 weeks. Before the rabbits were killed, the fracture sites were cultured to document the presence or absence of infection. The results of all infected osteotomy sites were positive, whereas no positive scans were found in the noninfected osteotomies. We concluded from this study that uncomplicated fracture healing does not result in a positive indium-111 leukocyte scan.

  2. Prionemia and leukocyte-platelet-associated infectivity in sheep transmissible spongiform encephalopathy models.

    PubMed

    Lacroux, Caroline; Vilette, Didier; Fernández-Borges, Natalia; Litaise, Claire; Lugan, Séverine; Morel, Nathalie; Corbičre, Fabien; Simon, Stéphanie; Simmons, Hugh; Costes, Pierrette; Weisbecker, Jean-Louis; Lantier, Isabelle; Lantier, Frederic; Schelcher, François; Grassi, Jacques; Castilla, Joaquin; Andréoletti, Olivier

    2012-02-01

    The dynamics of the circulation and distribution of transmissible spongiform encephalopathy (TSE) agents in the blood of infected individuals remain largely unknown. This clearly limits the understanding of the role of blood in TSE pathogenesis and the development of a reliable TSE blood detection assay. Using two distinct sheep scrapie models and blood transfusion, this work demonstrates the occurrence of a very early and persistent prionemia. This ability to transmit disease by blood transfusion was correlated with the presence of infectivity in white blood cells (WBC) and peripheral blood mononucleated cells (PBMC) as detected by bioassay in mice overexpressing the ovine prion protein PrP (tg338 mice) and with the identification of abnormal PrP in WBC after using protein misfolding cyclic amplification (PMCA). Platelets and a large variety of leukocyte subpopulations also were shown to be infectious. The use of endpoint titration in tg338 mice indicated that the infectivity in WBC (per ml of blood) was 10(6.5)-fold lower than that in 1 g of posterior brainstem sample. In both WBC and brainstem, infectivity displayed similar resistance to PK digestion. The data strongly support the concept that WBC are an accurate target for reliable TSE detection by PMCA. The presence of infectivity in short-life-span blood cellular elements raises the question of the origin of prionemia. PMID:22156536

  3. Pentoxifylline-induced modulation of human leukocyte function in vitro.

    PubMed Central

    Josaki, K.; Contrino, J.; Kristie, J.; Krause, P.; Kreutzer, D. L.

    1990-01-01

    We previously demonstrated that pentoxifylline stimulated leukocyte migration in vitro and leukocyte accumulation in vivo and protects neonatal mice from experimentally induced Staphylococcus aureus infections. In the present studies we have investigated pentoxifylline's effect on human leukocyte function in vitro. In these studies we demonstrate that pentoxifylline at low concentrations (ie, 0.01 and 0.1 mg/ml) stimulates both leukocyte migration and microbicidal activity in vitro. Alternatively, low concentrations (0.001 to 0.1 mg/ml) of pentoxifylline had no significant effect on the binding uptake of S. aureus by leukocytes, nor did it enhance phagocytic degranulation. At extremely low concentrations (0.001 mg/ml), pentoxifylline enhanced oxygen metabolism by human leukocytes, as reflected by increased H2O2 production and chemiluminescence (CL). At higher concentrations (ie, 0.1 to 1 mg/ml), pentoxifylline consistently suppressed these leukocyte functions in vitro. Thus, this study supports the following hypothesis: 1) the in vivo effects of pentoxifylline may involve a direct effect on both leukocyte mobilization and microbicidal activity, and 2) the enhanced microbicidal activity induced by pentoxifylline may be a result of enhanced leukocyte oxygen metabolism. In summary, pentoxifylline appears to be an interesting immunomodulator (ie, immunoenhancement and immunosuppression) of leukocyte function in vitro, but additional studies will be required before the efficacy of pentoxifylline in man can be determined. PMID:2316627

  4. Mouse carotid artery ligation induces platelet-leukocyte-dependent luminal fibrin, required for neointima development.

    PubMed

    Kawasaki, T; Dewerchin, M; Lijnen, H R; Vreys, I; Vermylen, J; Hoylaerts, M F

    2001-02-01

    The relationship between platelet and leukocyte activation, coagulation, and neointima development was investigated in noninjured murine blood vessels subjected to blood stasis. The left common carotid artery of C57BL/6J mice was ligated proximal to the bifurcation. Tissue-factor expression in luminal leukocytes progressively increased over 2 weeks. On day 3 after ligation, in addition to infiltrated granulocytes, platelet microthrombi and platelet-covered leukocytes as well as tissue-factor-positive fibrin deposits lined the endothelium. Maximal neointima formation in carotid artery cross sections of control mice equaled 28+/-3.7% (n=11) and 42+/-5.1% (n=8) of the internal elastic lamina cross-sectional area 1 and 2 weeks after ligation. In FVIII(-/-) mice, stenosis was significantly lower 1 (11+/-3.6%, n=8) and 2 (21+/-4.7%, n=7) weeks after ligation (both P:<0.01 versus background-matched controls). In u-PA(-/-) mice, luminal stenosis was significantly higher 1 (38+/-7.0%, n=7) and 2 (77+/-5.6%, n=6) weeks after ligation (P:<0.05 and P:<0.01, respectively, versus matched controls). In alpha(2)-AP(-/-) mice, stenosis was lower at 1 week (14+/-2.6%, n=7, P:<0.01) but not at 2 weeks. Responses in tissue-type plasminogen activator or plasminogen activator inhibitor-1 gene-deficient mice equaled that in controls. Reducing plasma fibrinogen levels in controls with ancrod or inducing partial thrombocytopenia with busulfan resulted in significantly less neointima, but inflammation was inhibited only in busulfan-treated mice. We conclude that stasis induces platelet activation, leading to microthrombosis and platelet-leukocyte conjugate formation, triggering inflammation and tissue-factor accumulation on the carotid artery endothelium. Delayed coagulation then results in formation of a fibrin matrix, which is used by smooth muscle cells to migrate into the lumen. PMID:11157667

  5. Leukocyte profiles for western fence lizards, Sceloporus occidentalis, naturally infected by the malaria parasite Plasmodium mexicanum.

    PubMed

    Motz, Victoria L; Lewis, William D; Vardo-Zalik, Anne M

    2014-10-01

    Plasmodium mexicanum is a malaria parasite that naturally infects the western fence lizard, Sceloporus occidentalis , in northern California. We set out to determine whether lizards naturally infected with this malaria parasite have different leukocyte profiles, indicating an immune response to infection. We used 29 naturally infected western fence lizards paired with uninfected lizards based on sex, snout-to-vent length, tail status, and the presence-absence of ectoparasites such as ticks and mites, as well as the presence-absence of another hemoparasite, Schellackia occidentalis. Complete white blood cell (WBC) counts were conducted on blood smears stained with Giemsa, and the proportion of granulocytes per microliter of blood was estimated using the Avian Leukopet method. The abundance of each WBC class (lymphocytes, monocytes, heterophils, eosinophils, and basophils) in infected and uninfected lizards was compared to determine whether leukocyte densities varied with infection status. We found that the numbers of WBCs and lymphocytes per microliter of blood significantly differed (P < 0.05) between the 2 groups for females but not for males, whereas parasitemia was significantly correlated with lymphocyte counts for males, but not for females. This study supports the theory that infection with P. mexicanum stimulates the lizard's immune response to increase the levels of circulating WBCs, but what effect this has on the biology of the parasite remains unclear. PMID:24945903

  6. Eosinophil Purification from Peripheral Blood

    PubMed Central

    Akuthota, Praveen; Capron, Kelsey; Weller, Peter F.

    2014-01-01

    Eosinophils are granulocytes integral to allergic inflammation and parasitic responses and comprise 1–4% of the circulating leukocytes in human beings under normal conditions. Isolation of human eosinophils allows for ex vivo and in vitro experimentation, providing a valuable tool for the study of allergic mechanisms. Here, we describe a technique for the isolation of human eosinophils by negative selection from whole blood obtained by venipuncture. PMID:24986603

  7. Chemiluminescence response of beta-glucan stimulated leukocytes isolated from different tissues and peritoneal cavity of Dicentrarchus labrax.

    PubMed

    Vazzana, Mirella; Parrinello, Daniela; Cammarata, Matteo

    2003-05-01

    The respiratory burst of leukocytes isolated from sea bass (Dicentrarchus labrax) pronephros, peritoneal cavity (P.C.), spleen and blood, was measured by a chemiluminescence (CL) assay after stimulation with beta-glucan. The CL response by P.C. and pronephros leukocytes was significantly higher than that expressed by a similar number of cells separated from spleen and blood. This probably reflects the observation that the proportion of macrophages and neutrophils was highest in the populations of leukocytes from peritoneal cavity and pronephros. Comparative observations showed a higher degree of yeast phagocytosis by leukocytes taken from peritoneal cavity than the pronephros. Moreover phagocytic index evaluated by microscopical observations, indicated that peritoneal macrophages internalised more yeast cells than neutrophils (identified by the peroxidase reaction). Scanning electron microscopy observations were also carried out. Inhibition experiments by a myeloperoxidase inhibitor sodium azide, iodonium-diphenyl-chloride which inhibits NADPH-oxidase, and exogenous superoxide dismutase, which catalyses O-2 dismutation to H(2)O(2), supported the correlation between CL and respiratory burst. Treatment with ouabain and DNP suggested that in this response, Ca(++) pump channels and calmodulin are involved in a metabolic energy-dependent pathway. PMID:12711275

  8. Leukocyte activation in advanced cancer as an explanation for absent leukocyte adherence inhibition to cancer extracts and chemoattractant.

    PubMed

    Labateya, N; Thomson, D M; Reid, E C

    1986-01-01

    Tube leukocyte adherence inhibition (LAI) measures human immunity by antigen-binding leukocytes releasing chemoattractant-like mediators that are the ultimate inhibitors of adherence by bystander leukocytes. We determined whether the absent LAI responses to cancer extracts for patients with large body burdens of bladder cancer was related to a defect in antigen binding or chemoattractant responsiveness. Leukocytes from patients with small body burdens of bladder cancer gave positive LAI responses of a similar extent to either bladder cancer extracts or chemoattractant [n-formyl-L-methionylleucylphenylalanine (FMLP)]. Of the adherent leukocytes, about 20-30% became non-adherent with a positive LAI response: monocytes, neutrophils and lymphocytes responded. In the control tubes, leukocytes from patients with large body burdens of bladder cancer were more non-adherent and about 15% less adherent than leukocytes from controls or patients with early cancer. They showed no further decrease in adherence, or conversely increase in non-adherence, with either extracts of bladder cancer or FMLP. The leukocytes also failed to transduce transmembrane signals to the same stimuli. The defect was reversible since PGE2 restored the adherence of leukocytes to normal, and subsequently they exhibited membrane potential changes and about 34% non-adherence to either bladder cancer extracts or FMLP. From these results we conclude that chemoattractant LAI-responsive leukocytes from patients with large body burdens of bladder tumor are activated in vivo, probably by mediators from inflammatory cells. PMID:3007156

  9. Carbon liberated from CO-releasing molecules attenuates leukocyte infiltration in the small intestine of thermally injured mice

    PubMed Central

    Sun, Bing-Wei; Jin, Qin; Sun, Yan; Sun, Zhi-Wei; Chen, Xi; Chen, Zhao-Yong; Cepinskas, Gediminas

    2007-01-01

    AIM: To determine whether Carbon (CO) liberated from CO-releasing molecules attenuates leukocyte infiltration in the small intestine of thermally injured mice. METHODS: Thirty-six mice were assigned to four groups. Mice in the sham group (n = 9) were underwent to sham thermal injury; mice in the burn group (n = 9) received 15% total body surface area full-thickness thermal injury; mice in the burn + CORM-2 group (n = 9) were underwent to the same thermal injury with immediate administration of tricarbonyldichlororuthenium (II) dimer CORM-2 (8 mg/kg, i.v.); and mice in the burn+DMSO group (n = 9) were underwent to the same thermal injury with immediate administration of 160 ?L bolus injection of 0.5% DMSO/saline. Histological alterations and granulocyte infiltration of the small intestine were assessed. Polymorphonuclear neutrophil (PMN) accumulation (myeloperoxidase assay) was assessed in mice mid-ileum. Activation of nuclear factor (NF)-??, expression levels of intercellular adhesion molecule-1 (ICAM-1) and inducible heme oxygenase in mid-ileum were assessed. RESULTS: Treatment of thermally injured mice with CORM-2 attenuated PMN accumulation and prevented activation of NF-?? in the small intestine. This was accompanied by a decrease in the expression of ICAM-1. In parallel, burn-induced granulocyte infiltration in mid-ileum was markedly decreased in the burn mice treated with CORM-2. CONCLUSION: CORM-released CO attenuates leukocyte infiltration in the small intestine of thermally injured mice by interfering with NF-?? activation and protein expression of ICAM-1, and therefore suppressing the pro-adhesive phenotype of endothelial cells. PMID:18069757

  10. Migration and chemiluminescence of polymorphonuclear cells and monocytes to Bacteroides sonicates.

    PubMed

    Fotos, P G; Lewis, D M; Gerencser, V F; Gerencser, M A; Snyder, I S

    1992-01-01

    Recent investigations have demonstrated that various preparations obtained from representatives of the genus Bacteroides are poorly phagocytized by polymorphonuclear cells (PMN) and macrophages. Crude cell sonicates derived from Bacteroides have been examined for their ability to inhibit migration of PMN and monocytes using a modified migration under agarose in vitro assay. B. gingivalis and B. intermedius were found to be inhibitors of such migration while B. asaccharolyticus did not share this property (P less than 0.005). In addition, B. intermedius sonicates were found to inhibit PMN chemiluminescence to known stimulants (P less than 0.001). These data were not found to result from direct sonicate cytotoxicity and therefore lend additional support to the etiologic importance of specific Bacteroides strains in the pathogenesis of acute and chronic dentoalveolar infections. PMID:1321583

  11. Human leukocyte antigen matching in renal transplantation: an update.

    PubMed

    Goes, N; Chandraker, A

    2000-11-01

    Cadaveric kidney allocation, in most countries, is based on human leukocyte antigen matching of the donor kidney with the recipient. Traditional human leukocyte antigen matching is based on defining human leukocyte antigen specificities by antibodies. Newer techniques have emerged from the tissue typing laboratory, which challenge the accuracy of serological typing and crossmatching. Improvements in renal allograft survival, predominantly as a result of newer immunosuppressive drugs, have led to longer survival times even in poorly matched human leukocyte antigen renal allografts. The scarcity of donor organs has focused attention on organ allocation policies, and the exact role of human leukocyte antigen matching in renal transplantation is under scrutiny. In this review, we examine developments in human leukocyte antigen matching as well as attempts to utilize this information to allocate cadaveric kidneys optimally. PMID:11128432

  12. Erythrocyte and blood antibacterial defense

    PubMed Central

    2014-01-01

    It is an axiom that blood cellular immunity is provided by leukocytes. As to erythrocytes, it is generally accepted that their main function is respiration. Our research provides objective video and photo evidence regarding erythrocyte bactericidal function. Phase-contrast immersion vital microscopy of the blood of patients with bacteremia was performed, and the process of bacteria entrapping and killing by erythrocytes was shot by means of video camera. Video evidence demonstrates that human erythrocytes take active part in blood bactericidal action and can repeatedly engulf and kill bacteria of different species and size. Erythrocytes are extremely important integral part of human blood cellular immunity. Compared with phagocytic leukocytes, the erythrocytes: a) are more numerous; b) are able to entrap and kill microorganisms repeatedly without being injured; c) are more resistant to infection and better withstand the attacks of pathogens; d) have longer life span and are produced faster; e) are inauspicious media for proliferation of microbes and do not support replication of chlamidiae, mycoplasmas, rickettsiae, viruses, etc.; and f) are more effective and uncompromised bacterial killers. Blood cellular immunity theory and traditional view regarding the function of erythrocytes in human blood should be revised. PMID:24883200

  13. Effect of elevated serum prolactin concentrations on the immunophenotype of human lymphocytes, mitogen-induced proliferation and phagocytic activity of polymorphonuclear cells.

    PubMed

    Köller, M; Kotzmann, H; Clodi, M; Riedl, M; Luger, A

    1997-08-01

    It has been suggested that the immune system is an important target tissue of prolactin (PRL). We therefore investigated several immune parameters in nine patients with chronically elevated serum prolactin concentrations. The immunophenotype of lymphocytes, mitogen-induced lymphocyte proliferation and phagocytic activity of polymorphonuclear cells were determined under high serum prolactin levels and 2 weeks after treatment with dopamine agonists. An increased CD4/CD8 ratio in the hyperprolactinaemic patients could be detected compared with healthy control subjects, which remained high after treatment and did not seem to correlate with serum prolactin concentrations. Peripheral blood B lymphocytes showed an increased expression of CD69 in the treated group but not in untreated patients compared with healthy control subjects. Interleukin 2 receptor, CD45RO, transferrin receptor or HLA-DR expression of CD4 or CD8 cells, as well as oxidative burst and phagocytic activity of granulocytes, were not affected in the patients with prolactinomas. Lymphocyte transformation response to phytohaemagglutinin in vitro was found not to be influenced by elevated prolactin levels except at the highest mitogen concentration tested. These data together with previous reports suggest that, although PRL is required for lymphocyte maturation to achieve normal immune function, elevated PRL levels do not lead to an 'overstimulation' of the immune system in men. PMID:9279529

  14. Leukocyte membrane bleb and pseudopod formation in hypertension

    PubMed Central

    Edwards, Kate M.; Sheu, Bryan; Hong, Suzi; Penn, Alexander H.; Schmid-Schönbein, Geert W.; Mills, Paul J

    2010-01-01

    Leukocyte activation, including adhesion molecule expression, oxygen radical generation and, in animal studies, pseudopod formation, is a hallmark of hypertension. This study examined pseudopod and bleb formation and demonstrates that leukocytes from hypertensive individuals are more susceptible to produce membrane blebs than leukocytes from normotensive individuals. Bleb formation is likely indicative of apoptosis, thus this observation adds to previous observations of increased apoptosis in various tissues in hypertension. PMID:20410916

  15. Tryptophan/kynurenine metabolism in human leukocytes is independent of superoxide and is fully maintained in chronic granulomatous disease.

    PubMed

    De Ravin, Suk See; Zarember, Kol A; Long-Priel, Debra; Chan, King C; Fox, Stephen D; Gallin, John I; Kuhns, Douglas B; Malech, Harry L

    2010-09-01

    In chronic granulomatous disease (CGD), defective phagocytic nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity causes reduced superoxide anion (O(2)(·)) radical production leading to frequent infections as well as granulomas and impaired wound healing indicative of excessive inflammation. Based on recent mouse studies, the lack of O(2)(·)-dependent interferon ? (IFN?)-induced synthesis of kynurenine (kyn), an anti-inflammatory tryptophan metabolite produced by indolamine 2,3 deoxygenase (IDO), was proposed as a cause of hyperinflammation in CGD and this pathway has been considered for clinical intervention. Here, we show that IFN? induces normal levels of kynurenine in cultures of O(2)(·)-deficient monocytes, dendritic cells, and polymorphonuclear leukocytes from gp91(PHOX)- or p47(PHOX)-deficient human CGD donors. Kynurenine accumulation was dose- and time-dependent as was that of a downstream metabolite, anthranilic acid. Furthermore, urinary and serum levels of kynurenine and a variety of other tryptophan metabolites were elevated rather than suppressed in CGD donors. Although we did not specifically evaluate kyn metabolism in local tissue or inflamed sites in humans, our data demonstrates that O(2)(·) anion is dispensable for the rate-limiting step in tryptophan degradation, and CGD patients do not appear to have either hematopoietic cell or systemic deficits in the production of the anti-inflammatory kynurenine molecule. PMID:20511543

  16. Tryptophan/kynurenine metabolism in human leukocytes is independent of superoxide and is fully maintained in chronic granulomatous disease

    PubMed Central

    Zarember, Kol A.; Long-Priel, Debra; Chan, King C.; Fox, Stephen D.; Gallin, John I.; Kuhns, Douglas B.; Malech, Harry L.

    2010-01-01

    In chronic granulomatous disease (CGD), defective phagocytic nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity causes reduced superoxide anion (O2??) radical production leading to frequent infections as well as granulomas and impaired wound healing indicative of excessive inflammation. Based on recent mouse studies, the lack of O2??-dependent interferon ? (IFN?)–induced synthesis of kynurenine (kyn), an anti-inflammatory tryptophan metabolite produced by indolamine 2,3 deoxygenase (IDO), was proposed as a cause of hyperinflammation in CGD and this pathway has been considered for clinical intervention. Here, we show that IFN? induces normal levels of kynurenine in cultures of O2??-deficient monocytes, dendritic cells, and polymorphonuclear leukocytes from gp91PHOX- or p47PHOX-deficient human CGD donors. Kynurenine accumulation was dose- and time-dependent as was that of a downstream metabolite, anthranilic acid. Furthermore, urinary and serum levels of kynurenine and a variety of other tryptophan metabolites were elevated rather than suppressed in CGD donors. Although we did not specifically evaluate kyn metabolism in local tissue or inflamed sites in humans, our data demonstrates that O2?? anion is dispensable for the rate-limiting step in tryptophan degradation, and CGD patients do not appear to have either hematopoietic cell or systemic deficits in the production of the anti-inflammatory kynurenine molecule. PMID:20511543

  17. Tumor-elicited polymorphonuclear cells, in contrast to "normal" circulating polymorphonuclear cells, stimulate invasive and metastatic potentials of rat mammary adenocarcinoma cells.

    PubMed Central

    Welch, D R; Schissel, D J; Howrey, R P; Aeed, P A

    1989-01-01

    Circulating polymorphonuclear cell (PMN) levels rise in proportion to the metastatic potential of the tumor in 13762NF mammary adenocarcinoma tumor-bearing rats. These tumor-elicited PMNs (tcPMNs) secrete high levels of the basement-membrane-degrading enzymes, type IV collagenase and heparanase, suggesting that metastatic tumor cells stimulate neutrophilia so that the tcPMNs might assist tumor cell extravasation during metastasis. To test this hypothesis, purified proteose peptone-elicited PMNs from peritoneal exudate, circulating normal PMNs, and tcPMNs were evaluated for their effects on in vitro invasive and in vivo metastatic potentials of syngeneic 13762NF mammary adenocarcinoma tumor cells. tcPMNs caused a dose-dependent increase in invasion through a reconstituted basement membrane barrier in an in vitro invasion assay. At PMN:tumor cell ratios of 30:1, invasion potential significantly (P less than 0.05) rose to 26-fold, 40-fold, and 37-fold for poorly metastatic MTLn2 cells, highly metastatic MTLn3 cells, and moderately metastatic MTF7 cells, respectively. In contrast, purified proteose peptone-elicited PMNs and circulating normal PMNs did not significantly alter invasive potential. Intravenous coinjections of purified proteose peptone-elicited PMNs did not change the number of experimental lung metastases, but tcPMNs at ratios to 50:1 significantly raised the mean number of metastases 23-fold for MTLn2, 3- to 4-fold for MTLn3, and 1.6- to 1.8-fold for MTF7. These results demonstrate that tcPMNs contribute to the metastatic propensity of mammary adenocarcinoma clones by increasing efficiency of invasion through basement membrane. PMID:2762301

  18. Cerebral nitric oxide represses choroid plexus NF?B-dependent gateway activity for leukocyte trafficking.

    PubMed

    Baruch, Kuti; Kertser, Alexander; Porat, Ziv; Schwartz, Michal

    2015-07-01

    Chronic neuroinflammation is evident in brain aging and neurodegenerative disorders and is often associated with excessive nitric oxide (NO) production within the central nervous system (CNS). Under such conditions, increased NO levels are observed at the choroid plexus (CP), an epithelial layer that forms the blood-cerebrospinal fluid barrier (BCSFB) and serves as a selective gateway for leukocyte entry to the CNS in homeostasis and following injury. Here, we hypothesized that elevated cerebral NO levels interfere with CP gateway activity. We found that induction of leukocyte trafficking determinants by the CP and sequential leukocyte entry to the CSF are dependent on the CP epithelial NF?B/p65 signaling pathway, which was inhibited upon exposure to NO. Examining the CP in 5XFAD transgenic mouse model of Alzheimer's disease (AD-Tg) revealed impaired ability to mount an NF?B/p65-dependent response. Systemic administration of an NO scavenger in AD-Tg mice alleviated NF?B/p65 suppression at the CP and augmented its gateway activity. Together, our findings identify cerebral NO as a negative regulator of CP gateway activity for immune cell trafficking to the CNS. PMID:25940071

  19. Genotoxic potential of TiO2 on bottlenose dolphin leukocytes.

    PubMed

    Bernardeschi, Margherita; Guidi, Patrizia; Scarcelli, Vittoria; Frenzilli, Giada; Nigro, Marco

    2010-01-01

    Titanium dioxide is extensively used in a variety of products, including industrial materials and cosmetics. Studies mainly performed on human cell lines and in vivo exposure on experimental animals have raised concern about the toxic effects of ultrafine titanium dioxide; however, scarce information is available about its impact on aquatic life. The aim of this article was to assess the genotoxic potential of TiO(2) (anatase and rutile) on bottlenose dolphin leukocytes. Blood samples were obtained from four male and one female specimens reared at the Adriatic SeaWorld "Oltremare" (Riccione, Italy). Leukocytes were isolated by the lyses procedure and in vitro exposed to TiO(2) in RPMI. Experimental solutions were sonicated immediately before dosing the cells. Three exposure times (4, 24 and 48 h) and three doses (20, 50 and 100 microg/ml) were tested. Genotoxicity was detected by the single-cell gel electrophoresis (or comet assay) at pH > or = 13, assessing single/double-strand breaks and alkali-labile sites. Cytotoxicity was also detected by the Trypan blue exclusion method. Results showed that both the crystalline forms of TiO(2) were genotoxic for bottlenose dolphin leukocytes, with a statistically significant increase of DNA fragmentation after exposure to 50 and 100 microg/ml for 24 and 48 h. Although preliminary, these are the first data regarding the genetic susceptibility of toothed cetaceans toward an "emerging" pollutant, such as TiO(2) particles. PMID:19915826

  20. Childhood Physical and Sexual Abuse History and Leukocyte Telomere Length among Women in Middle Adulthood

    PubMed Central

    Mason, Susan M.; Prescott, Jennifer; Tworoger, Shelley S.; DeVivo, Immaculata; Rich-Edwards, Janet W.

    2015-01-01

    Objective Abuse victimization in childhood is associated with a variety of age-related cardiometabolic diseases, but the mechanisms remain unknown. Telomeres, which form the protective caps at the ends of chromosomes, have been proposed as measures of biological age, and a growing body of research suggests that telomere attrition may help to explain relationships between stress and cardiometabolic degradation. We examined the association between childhood abuse victimization and leukocyte telomere length among 1,135 participants in the Nurses’ Health Study II (NHSII). Methods The NHSII ascertained physical and sexual child abuse histories in 2001. Telomere length was measured in genomic DNA extracted from peripheral blood leukocytes collected between 1996 and 1999. The ratio of telomere repeat copy number to a single gene copy number (T/S) was determined by a modified version of the quantitative real-time PCR telomere assay. Telomere length was log-transformed and corrected for assay variation across batch. We regressed telomere length on childhood abuse exposure variables and covariates using linear regression. Results We observed a reduction in telomere length associated with moderate physical abuse versus no physical abuse, but there was no evidence of a dose-response relationship for increased severity of physical abuse. No associations were noted for sexual abuse. Conclusions We found no evidence of an association between severity of childhood physical or sexual abuse and leukocyte telomere length in the NHSII. PMID:26053088

  1. Proteomic analysis of peripheral leukocytes in Alzheimer’s disease patients treated with divalproex sodium

    PubMed Central

    Mhyre, Timothy R.; Loy, Rebekah; Tariot, Pierre N.; Profenno, Louis A.; Maguire-Zeiss, Kathleen A.; Zhang, Dabao; Coleman, Paul D.; Federoff, Howard J.

    2008-01-01

    The molecular profiling of peripheral tissues, including circulating leukocytes, may hold promise in the discovery of biomarkers for diagnosing and treating neurodegenerative diseases, including Alzheimer’s disease (AD). As a proof-of-concept, we performed a proteomics study on peripheral leukocytes from patients with AD both before and during treatment with divalproex sodium. Using two-dimensional gel electrophoresis and MALDI-TOF mass spectrometry, we identified ten differentially expressed proteins: two up-regulated proteins, 14-3-3 protein ? and peroxiredoxin 2; and eight down-regulated proteins, actin-interacting protein, mitogen activated protein kinase 1, beta actin, annexin A1, glyceraldehyde 3-phosphate dehydrogenase, transforming protein RhoA, acidic leucine-rich nuclear phosphoprotein 32 family member B, and a currently unidentified protein. A subset was validated on both the transcript and protein levels in normal human peripheral blood mononuclear cell cultures treated with valproic acid. These proteins comprise a number of functional classes that may be important to the biology of AD and to the therapeutic action of valproate. These data also suggest the potential of using peripheral leukocytes to monitor pharmaceutical action for neurodegenerative diseases. PMID:17521776

  2. Umbilical cord blood transplantation.

    PubMed

    Koo, Hong Hoe; Ahn, Hyo Seop

    2012-07-01

    Since the first umbilical cord blood transplantation (CBT) in 1998, cord blood (CB) has now become one of the most commonly used sources of hematopoietic stem cells for transplantation. CBT has advantages of easy procurement, no risk to donor, low risk of transmitting infections, immediate availability and immune tolerance allowing successful transplantation despite human leukocyte antigen disparity. Several studies have shown that the number of cells transplanted is the most important factor for engraftment in CBT, and it limits the wide use of CB in adult patients. New strategies for facilitating engraftment and reducing transplantation-related mortality are ongoing in the field of CBT and include the use of a reduced-intensity conditioning regimen, double-unit CBT, ex vivo expansion of CB, and co-transplantation of CB and mesenchymal stem cells. Recently, the results of two international studies with large sample sizes showed that CB is an acceptable alternative source of hematopoietic stem cells for adult recipients who lack human leukocyte antigen-matched adult donors. Along with the intensive researches, development in banking process of CB will amplify the use of CB and offer the chance for cure in more patients. PMID:22844315

  3. Umbilical cord blood transplantation

    PubMed Central

    Koo, Hong Hoe

    2012-01-01

    Since the first umbilical cord blood transplantation (CBT) in 1998, cord blood (CB) has now become one of the most commonly used sources of hematopoietic stem cells for transplantation. CBT has advantages of easy procurement, no risk to donor, low risk of transmitting infections, immediate availability and immune tolerance allowing successful transplantation despite human leukocyte antigen disparity. Several studies have shown that the number of cells transplanted is the most important factor for engraftment in CBT, and it limits the wide use of CB in adult patients. New strategies for facilitating engraftment and reducing transplantation-related mortality are ongoing in the field of CBT and include the use of a reduced-intensity conditioning regimen, double-unit CBT, ex vivo expansion of CB, and co-transplantation of CB and mesenchymal stem cells. Recently, the results of two international studies with large sample sizes showed that CB is an acceptable alternative source of hematopoietic stem cells for adult recipients who lack human leukocyte antigen-matched adult donors. Along with the intensive researches, development in banking process of CB will amplify the use of CB and offer the chance for cure in more patients. PMID:22844315

  4. Regulated Expression of Human Histocompatibility Leukocyte Antigen (HLA)DO During Antigen-dependent and Antigen-independent Phases of B Cell Development

    Microsoft Academic Search

    Xinjian Chen; Oskar Laur; Taku Kambayashi; Shiyong Li; Robert A. Bray; Dominique A. Weber; Lars Karlsson; Peter E. Jensen

    2002-01-01

    Human histocompatibility leukocyte antigen (HLA)-DO, a lysosomal resident major histo- compatibility complex class II molecule expressed in B cells, has previously been shown to be a negative regulator of HLA-DM peptide loading function. We analyze the expression of DO in human peripheral blood, lymph node, tonsil, and bone marrow to determine if DO expression is modulated in the physiological setting.

  5. Cytokine Production by Leukocytes of Military Personnel with Depressive Symptoms after Deployment to a Combat-Zone: A Prospective, Longitudinal Study

    Microsoft Academic Search

    Mirjam van Zuiden; Cobi J. Heijnen; Rens van de Schoot; Karima Amarouchi; Mirjam Maas; Eric Vermetten; Elbert Geuze; Annemieke Kavelaars

    2011-01-01

    Major depressive disorder (MDD) is frequently diagnosed in military personnel returning from deployment. Literature suggests that MDD is associated with a pro-inflammatory state. To the best of our knowledge, no prospective, longitudinal studies on the association between development of depressive symptomatology and cytokine production by peripheral blood leukocytes have been published. The aim of this study was to investigate whether

  6. CD11c expression in adipose tissue and blood and its role in diet-induced obesity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To examine CD11c, a beta(2)-integrin, on adipose tissue (AT) leukocytes, and blood monocytes and its role in diet-induced obesity. High-fat diet-induced obese C57BL/6 mice, CD11c-deficient mice, and obese humans were studied. CD11c, leukocytes, and chemokines/cytokines were examined in AT and/or blo...

  7. INHIBITION OF TUMOUR NECROSIS FACTOR PRODUCTION IN ENDOTOXIN-STIMULATED HUMAN MONONUCLEAR LEUKOCYTES BY THE PROSTACYCLIN ANALOGUE ILOPROST: CELLULAR MECHANISMS

    Microsoft Academic Search

    Achim Jörres; Harald Dinter; Nicholas Topley; Gerhard M. Gahl; Ulrich Frei; Peter Scholz

    1997-01-01

    The prostacyclin analogue iloprost has been shown to inhibit effectively TNF-? production in human peripheral blood mononuclear leukocytes (PBMC) stimulated with bacterial lipopolysaccharide (LPS). The current paper set out to analyse further the possible mechanisms involved in the regulation of TNF-? synthesis by iloprost. Healthy human PBMC were challenged withEscherichia coliLPS and assessed for TNF-? gene transcription, mRNA stability and

  8. The effect of (1?3)-?- d-glucans, carboxymethylglucan and schizophyllan on human leukocytes in vitro

    Microsoft Academic Search

    Lukas Kubala; Jana Ruzickova; Kristina Nickova; Jozef Sandula; Milan Ciz; Antonin Lojek

    2003-01-01

    (1?3)-?-d-glucans are known as potent inductors of humoral and cell-mediated immunity in humans and animals. (1?3)-?-d-glucans isolated from various sources differ in their chemical structure and physical parameters and consequently in their immunomodulatory potential. In this study the immunomodulatory activity of two (1?3)-?-d-glucans schizophyllan (SPG) and carboxymethylglucan (CMG) was determined and compared on human blood leukocytes in vitro. Both SPG

  9. Fundamentals of the psoralen-based Helinx ™ technology for inactivation of infectious pathogens and leukocytes in platelets and plasma

    Microsoft Academic Search

    Susan Wollowitz

    2001-01-01

    Psoralens plus ultraviolet A (UVA) light inactivate viruses and bacteria as well as leukocytes. A system employing the synthetic psoralen compound amotosalen hydrochloride (S-59), in combination with UVA light, is being developed to decontaminate platelet concentrates and plasma in a blood-bank setting. S-59 is a heterocyclic psoralen compound that reacts by a three-step process with nucleic acids (NAs): (1) S-59

  10. Fluorescence spectroscopic detection of mitochondrial flavoprotein redox oscillations and transient reduction of the NADPH oxidase-associated flavoprotein in leukocytes

    Microsoft Academic Search

    Andrei Kindzelskii; Howard R. Petty

    2004-01-01

    Steady-state and time-resolved fluorescence spectroscopy and fluorescence microscopy of leukocyte flavoproteins have been performed. Both living human peripheral blood monocytes and neutrophils have been utilized as experimental models, as the former relies much more heavily on mitochondrial metabolism for energy production than the latter. We confirm previous studies indicating that cellular flavoproteins absorb at 460 nm and emit at 530 nm, very

  11. Medical Laboratory Technician--Hematology, Serology, Blood Banking, and Immunohematology (AFSC 90470).

    ERIC Educational Resources Information Center

    Thompson, Joselyn H.

    This three-volume student text is designed for use by Air Force personnel enrolled in a self-study extension course for medical laboratory technicians. Covered in the individual volumes are hematology (the physiology of blood, complete blood counts and related studies, erythrocyte studies, leukocyte and thrombocyte maturation, and blood

  12. Blood Culture and Stimulation Conditions for the Diagnosis of Tuberculosis in Cervids by the Cervigam Assay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mitogen and antigen induced interferon-gamma (IFN-gamma) responses of peripheral blood leukocytes from cervids were evaluated using a commercial, whole blood assay for the cytokine (Cervigam trademark, Prionics AG). Whole blood was from Mycobacterium bovis-infected white-tailed deer and reindeer, M....

  13. Orally bioavailable benzisothiazolone inhibitors of human leukocyte elastase.

    PubMed

    Hlasta, D J; Subramanyam, C; Bell, M R; Carabateas, P M; Court, J J; Desai, R C; Drozd, M L; Eickhoff, W M; Ferguson, E W; Gordon, R J

    1995-03-01

    Human leukocyte elastase (HLE) has been proposed as a primary mediator of pulmonary emphysema and other inflammatory airway diseases. HLE is capable of cleaving many proteins, including elastin, other components of connective tissue, certain complement proteins, and receptors. Under normal conditions an appropriate balance exists in the lung between HLE and endogenous inhibitors, which scavenge the released enzyme before it exerts deleterious effects in the lung. Emphysema is thought to result from an imbalance in the lung between HLE and endogenous inhibitor (elevated elastase or insufficient inhibitor) that leads to the destruction of alveoli. We have identified WIN 64733 (2) and WIN 63759 (3) as potent (Ki* = 14 and 13 pM, respectively), selective, mechanism-based inhibitors of HLE which are orally bioavailable in the dog (absolute bioavailability 46% and 21%, respectively). In this series the in vitro stabilities of the inhibitors in blood, jejunal homogenates, and liver S9 homogenates are useful predictors of oral bioavailability. After being administered orally (30 mg/kg) to dogs, compounds 2 and 3 are found in the lung, being detected in the epithelial lining fluid obtained by bronchoalveolar lavage (Cmax of 2.5 and 0.47 microgram/mL, respectively). PMID:7877139

  14. Unsupervised explorative data analysis of normal human leukocytes and BCR/ABL positive leukemic cells mid-infrared spectra.

    PubMed

    Bellisola, G; Bolomini Vittori, M; Cinque, G; Dumas, P; Fiorini, Z; Laudanna, C; Mirenda, M; Sandt, C; Silvestri, G; Tomasello, L; Vezzalini, M; Wehbe, K; Sorio, C

    2015-06-15

    We proved the ability of Fourier Transform Infrared microspectroscopy (microFTIR) complemented by Principal Component Analysis (PCA) to detect protein phosphorylation/de-phosphorylation in mammalian cells. We analyzed by microFTIR human polymorphonuclear neutrophil (PMNs) leukocytes, mouse-derived parental Ba/F3 cells (Ba/F3#PAR), Ba/F3 cells transfected with p210(BCR/ABL) (Ba/F3#WT) and expressing high levels of protein tyrosine kinase (PTK), and human-derived BCR/ABL positive K562 leukemic cell sub-clones engineered to differently express receptor-type tyrosine-protein phosphatase gamma (PTPRG). Synchrotron radiation (SR) and conventional (globar) IR sources were used to perform microFTIR respectively, on single cells and over several cells within the same sample. Ex vivo time-course experiments were run, inducing maximal protein phosphorylation in PMNs by 100 nM N-formylated tripeptide fMLP. Within the specific IR fingerprint 1800-850 cm(-1) frequency domain, PCA identified two regions with maximal signal variance. These were used to model and test the robustness of PCA in representing the dynamics of protein phosphorylation/de-phosphorylation processes. An IR signal ratio marker reflecting the homeostatic control by protein kinases and phosphatases was identified in normal leukocytes. The models identified by microFTIR and PCA in normal leukocytes also distinguished BCR/ABL positive Ba/F3#WT from BCR/ABL negative Ba/F3#PAR cells as well as K562 cells exposed to functionally active protein tyrosine phosphatase recombinant protein ICD-Tat transduced in cells by HIV-1 Tat technology or cells treated with the PTK inhibitor imatinib mesylate (IMA) from cells exposed to phosphatase inactive (D1028A)ICD-Tat recombinant protein and untreated control cells, respectively. The IR signal marker correctly reflected the degrees of protein phosphorylation associated with abnormal PTK activity in BCR/ABL positive leukemic cells and in general was inversely related to the expression/activity of PTPRG in leukemic sub-clones. In conclusion, we have described a new, reliable and simple spectroscopic method to study the ex vivo protein phosphorylation/de-phosphorylation balance in cell models: it is suitable for biomedical and pharmacological research labs but it also needs further optimization and its evaluation on large cohorts of patients to be proposed in the clinical setting of leukemia. PMID:25988195

  15. Blood Disorders

    MedlinePLUS

    ... blood cells, white blood cells and platelets. Blood disorders affect one or more parts of the blood ... They can be acute or chronic. Many blood disorders are inherited. Other causes include other diseases, side ...

  16. Blood Transfusions

    MedlinePLUS

    ... their blood . Donors give blood at local blood banks, at community centers during blood drives, or through ... in the world. Many organizations, including community blood banks and the federal government, work hard to ensure ...

  17. Blood Tests

    MedlinePLUS

    ... t have serious reactions to having blood drawn. Laboratory (lab) workers draw the blood and analyze it. They use either whole blood to count blood cells, or they separate the blood cells from the ...

  18. Inhibition of junctional adhesion molecule-A/LFA interaction attenuates leukocyte trafficking and inflammation in brain ischemia/reperfusion injury

    PubMed Central

    Sladojevic, Nikola; Stamatovic, Svetlana M.; Keep, Richard F.; Grailer, Jamison J.; Sarma, J. Vidya; Ward, Peter A.; Andjelkovic, Anuska V.

    2014-01-01

    Proinflammatory mediators trigger intensive postischemic inflammatory remodeling of the blood–brain barrier (BBB) including extensive brain endothelial cell surface and junctional complex changes. Junctional adhesion molecule-A (JAM-A) is a component of the brain endothelial junctional complex with dual roles: paracellular route occlusion and regulating leukocyte docking and migration. The current study examined the contribution of JAM-A to the regulation of leukocyte (neutrophils and monocytes/macrophages) infiltration and the postischemic inflammatory response in brain ischemia/reperfusion (I/R injury). Brain I/R injury was induced by transient middle cerebral artery occlusion (MCAO) for 30 min in mice followed by reperfusion for 0–5 days, during which time JAM-A antagonist peptide (JAM-Ap) was administered. The peptide, which inhibits JAM-A/leukocyte interaction by blocking the interaction of the C2 domain of JAM-A with LFA on neutrophils and monocytes/macrophages, attenuated I/R-induced neutrophil and monocyte infiltration into brain parenchyma. Consequently, mice treated with JAM-A peptide during reperfusion had reduced expression (~3-fold) of inflammatory mediators in the ischemic penumbra, reduced infarct size (94 ± 39 vs 211 ± 38 mm3) and significantly improved neurological score. BBB hyperpermeability was also reduced. Collectively, these results indicate that JAM-A has a prominent role in regulating leukocyte infiltration after brain I/R injury and could be a new target in limiting post-ischemic inflammation. PMID:24657919

  19. Leukocyte-inspired biodegradable particles that selectively and avidly adhere to inflamed endothelium in vitro and in vivo

    NASA Astrophysics Data System (ADS)

    Sakhalkar, Harshad S.; Dalal, Milind K.; Salem, Aliasger K.; Ansari, Ramin; Fu, Jie; Kiani, Mohammad F.; Kurjiaka, David T.; Hanes, Justin; Shakesheff, Kevin M.; Goetz, Douglas J.

    2003-12-01

    We exploited leukocyte-endothelial cell adhesion chemistry to generate biodegradable particles that exhibit highly selective accumulation on inflamed endothelium in vitro and in vivo. Leukocyte-endothelial cell adhesive particles exhibit up to 15-fold higher adhesion to inflamed endothelium, relative to noninflamed endothelium, under in vitro flow conditions similar to that present in blood vessels, a 6-fold higher adhesion to cytokine inflamed endothelium relative to non-cytokine-treated endothelium in vivo, and a 10-fold enhancement in adhesion to trauma-induced inflamed endothelium in vivo due to the addition of a targeting ligand. The leukocyte-inspired particles have adhesion efficiencies similar to that of leukocytes and were shown to target each of the major inducible endothelial cell adhesion molecules (E-selectin, P-selectin, vascular cell adhesion molecule 1, and intercellular adhesion molecule 1) that are up-regulated at sites of pathological inflammation. The potential for targeted drug delivery to inflamed endothelium has significant implications for the improved treatment of an array of pathologies, including cardiovascular disease, arthritis, inflammatory bowel disease, and cancer.

  20. Human leukocyte antigens in inhabitants of Taiwan.

    PubMed

    Lee, C J; Lee, P C; Tai, J J; Lee, P H

    1990-07-01

    Human leukocyte antigens (HLA) were investigated in 200 healthy Taiwan inhabitants of Taiwanese (46 persons), Hakka (36), Tai-Ya Tribe (28) Pu-Long Tribe (30) Pai-Wan Tribe (30) and Lu-kai (30) descent. In those of Taiwanese and Hakka descent, HLA-A11 and A2 were the two most frequently seen human leukocyte antigens. HLA-A11 was seen in 69.6% in those of Taiwanese descent and 61.1% in those of Hakka descent. HLA-A2 was seen in 50.0% of the Taiwanese and 44.4% of the Hakka. For the B loci, B40 (Taiwanese, 43.5%; Hakka, 44.4%), Bw46 (Taiwanese, 28.3%; Hakka, 19.4%) and B13 (Taiwanese, 15.2%; Hakka, 30.6%) were the most common antigens. For the C loci, Cw3 (Taiwanese, 57.1%; Hakka 64.9%), Cw1 (Taiwanese, 28.6%; Hakka, 35.1%) and Cw7 (Taiwanese, 40.5%; Hakka, 29.9%) were commonly seen antigens. The most commonly detected antigens for the DR loci were: DR2 (Taiwanese, 45.1%; Hakka, 56.8%) and DR4 (Taiwanese, 37.3%; Hakka, 32.4%). These results disclose that no major human leukocyte antigenic differences exist between the two aforementioned groups in this study. In addition, there are no major differences in Taiwanese and Hakka descent, and those of south and north mainland Chinese descent. All bear some antigenic similarities. However, for aborigines, the original inhabitants of Taiwan, the HLA antigens are significantly different from the Han Chinese as a whole. Characteristic features are seen in A24, Bw60, DRw6 and DRw8. In the meantime, a high frequency of common antigens are shared among the tribes and consequently a large number of blank alleles are seen in the aborigines. This reflects the homozygosity effect which is often seen in an isolated society. PMID:1979593

  1. Leukocyte activity is altered in a ground based murine model of microgravity and proton radiation exposure.

    PubMed

    Sanzari, Jenine K; Romero-Weaver, Ana L; James, Gabrielle; Krigsfeld, Gabriel; Lin, Liyong; Diffenderfer, Eric S; Kennedy, Ann R

    2013-01-01

    Immune system adaptation during spaceflight is a concern in space medicine. Decreased circulating leukocytes observed during and after space flight infer suppressed immune responses and susceptibility to infection. The microgravity aspect of the space environment has been simulated on Earth to study adverse biological effects in astronauts. In this report, the hindlimb unloading (HU) model was employed to investigate the combined effects of solar particle event-like proton radiation and simulated microgravity on immune cell parameters including lymphocyte subtype populations and activity. Lymphocytes are a type of white blood cell critical for adaptive immune responses and T lymphocytes are regulators of cell-mediated immunity, controlling the entire immune response. Mice were suspended prior to and after proton radiation exposure (2 Gy dose) and total leukocyte numbers and splenic lymphocyte functionality were evaluated on days 4 or 21 after combined HU and radiation exposure. Total white blood cell (WBC), lymphocyte, neutrophil, and monocyte counts are reduced by approximately 65%, 70%, 55%, and 70%, respectively, compared to the non-treated control group at 4 days after combined exposure. Splenic lymphocyte subpopulations are altered at both time points investigated. At 21 days post-exposure to combined HU and proton radiation, T cell activation and proliferation were assessed in isolated lymphocytes. Cell surface expression of the Early Activation Marker, CD69, is decreased by 30% in the combined treatment group, compared to the non-treated control group and cell proliferation was suppressed by approximately 50%, compared to the non-treated control group. These findings reveal that the combined stressors (HU and proton radiation exposure) result in decreased leukocyte numbers and function, which could contribute to immune system dysfunction in crew members. This investigation is one of the first to report on combined proton radiation and simulated microgravity effects on hematopoietic, specifically immune cells. PMID:23977138

  2. Leukocyte Activity Is Altered in a Ground Based Murine Model of Microgravity and Proton Radiation Exposure

    PubMed Central

    Sanzari, Jenine K.; Romero-Weaver, Ana L.; James, Gabrielle; Krigsfeld, Gabriel; Lin, Liyong; Diffenderfer, Eric S.; Kennedy, Ann R.

    2013-01-01

    Immune system adaptation during spaceflight is a concern in space medicine. Decreased circulating leukocytes observed during and after space flight infer suppressed immune responses and susceptibility to infection. The microgravity aspect of the space environment has been simulated on Earth to study adverse biological effects in astronauts. In this report, the hindlimb unloading (HU) model was employed to investigate the combined effects of solar particle event-like proton radiation and simulated microgravity on immune cell parameters including lymphocyte subtype populations and activity. Lymphocytes are a type of white blood cell critical for adaptive immune responses and T lymphocytes are regulators of cell-mediated immunity, controlling the entire immune response. Mice were suspended prior to and after proton radiation exposure (2 Gy dose) and total leukocyte numbers and splenic lymphocyte functionality were evaluated on days 4 or 21 after combined HU and radiation exposure. Total white blood cell (WBC), lymphocyte, neutrophil, and monocyte counts are reduced by approximately 65%, 70%, 55%, and 70%, respectively, compared to the non-treated control group at 4 days after combined exposure. Splenic lymphocyte subpopulations are altered at both time points investigated. At 21 days post-exposure to combined HU and proton radiation, T cell activation and proliferation were assessed in isolated lymphocytes. Cell surface expression of the Early Activation Marker, CD69, is decreased by 30% in the combined treatment group, compared to the non-treated control group and cell proliferation was suppressed by approximately 50%, compared to the non-treated control group. These findings reveal that the combined stressors (HU and proton radiation exposure) result in decreased leukocyte numbers and function, which could contribute to immune system dysfunction in crew members. This investigation is one of the first to report on combined proton radiation and simulated microgravity effects on hematopoietic, specifically immune cells. PMID:23977138

  3. Effect of complement and of the carbohydrate components of sputum on phagocytosis by human polymorphonuclear leucocytes

    PubMed Central

    Brogan, T. D.

    1964-01-01

    The phagocytic activity of human polymorphonuclear leucocyte preparations, which were free from plasma, has been estimated by direct determination under phase contrast of the number of living cells containing test particles. Spores of Aspergillus fumigatus were phagocytosed in the absence of added serum but phagocytosis of paraffin wax particles occurred only in the presence of serum containing the heat-labile and C?4 components of complement. In view of the unreactive nature of the paraffin hydrocarbons, it was considered unlikely that natural antibody played any part in the phenomenon. Although no phagocytosis of wax particles occurred in the absence of serum, almost 100 per cent of cells were phagocytic in preparations containing adequate concentrations of serum. It was therefore possible to determine the serum concentration necessary for 50 per cent of the polymorphs to phagocytose wax particles. By this means it was demonstrated that the addition of the carbohydrate components of sputum had a small but significant inhibitory effect on phagocytosis and that dextran had no such effect. The sputum mucoprotein depressed the complement titre of serum and this might have accounted for the reduction in the ability of a serum to promote phagocytosis when this complex was added. The sputum mucopolysaccharide had no such effect on the complement titre of serum and must have exerted its inhibitory action in some other way. ImagesFIG. 2 PMID:14239838

  4. Polymorphonuclear Cell Functional Impairment in Relapsing Remitting Multiple Sclerosis Patients: Preliminary Data

    PubMed Central

    Rossi, Silvia; Musso, Tiziana; Crocillŕ, Cristina; Cavalla, Paola; Trebini, Claudia; Marra, Elisa Simona; Cuffini, Anna Maria; Banche, Giuliana

    2015-01-01

    Multiple Sclerosis patients run an increased risk of microbial infections, which leads to high rates of hospitalization and infection-related mortality. Although immunotherapy may increase infection risk in some cases, data as to the relationship among microbial factors, immunotherapy and alterations in the innate immunity of these patients are still scanty. On these grounds, this interdisciplinary study aims at investigating the role the functional activity of polymorphonuclear cells (PMNs) play in relapsing remitting multiple sclerosis at different stages. The in vitro ability of PMNs from patients, either untreated or treated with immunosuppressant or immunomodulatory drugs to kill Klebsiella pneumonia or Candida albicans, were investigated and compared to PMNs from healthy subjects. The release of various cytokines was also assessed, as was the production of reactive oxygen species and their ability to regulate apoptosis after microbial stimulation. Our results indicate that although patients have a normal number of PMNs, they have a statistically significant (p<0.05) reduction in intracellular killing activity. Although variations are strongly related to the therapeutic management of patients, they are independent from their disease stage. As no statistically significant differences were observed between patients and controls in cytokine release values, reactive oxygen species production or apoptosis, we came to the conclusion that other factors may be involved. Supportive validation of these results from further studies might well help in identifying a subset of patients at high risk of infection who could benefit from a closer follow-up and/or antibiotic prophylaxis. PMID:26121651

  5. The beetroot component betanin modulates ROS production, DNA damage and apoptosis in human polymorphonuclear neutrophils.

    PubMed

    Zieli?ska-Przyjemska, Ma?gorzata; Olejnik, Anna; Kostrzewa, Artur; ?uczak, Micha?; Jagodzi?ski, Pawe? P; Baer-Dubowska, Wanda

    2012-06-01

    The aim of this study was to evaluate the effect of betanin, one of the beetroot major components, on ROS production, DNA damage and apoptosis in human resting and stimulated with phorbol 12-myristate13-acetate polymorphonuclear neutrophils, one of the key elements of the inflammatory response. Incubation of neutrophils with betanin in the concentration range 2-500?µM resulted in significant inhibition of ROS production (by 15-46%, depending on the ROS detection assay). The antioxidant capacity of betanin was most prominently expressed in the chemiluminescence measurements. This compound decreased also the percentage of DNA in comet tails in stimulated neutrophils, but only at the 24?h time point. In resting neutrophils an increased level of DNA in comet tails was observed. Betanin did not affect the activity of caspase-3, in resting neutrophils, but significantly enhanced the enzyme activity in stimulated neutrophils. The western blot analysis showed, however, an increased level of caspase-3 cleavage products as a result of betanin treatment both in resting and stimulated neutrophils. The results indicate that betanin may be responsible for the effect of beetroot products on neutrophil oxidative metabolism and its consequences, DNA damage and apoptosis. The dose and time dependent effects on these processes require further studies. PMID:22076941

  6. CD66 carcinoembryonic antigens mediate interactions between Opa-expressing Neisseria gonorrhoeae and human polymorphonuclear phagocytes.

    PubMed Central

    Gray-Owen, S D; Dehio, C; Haude, A; Grunert, F; Meyer, T F

    1997-01-01

    Colonization of urogenital tissues by the human pathogen Neisseria gonorrhoeae is characteristically associated with purulent exudates of polymorphonuclear phagocytes (PMNs) containing apparently viable bacteria. Distinct variant forms of the phase-variable opacity-associated (Opa) outer membrane proteins mediate the non-opsonized binding and internalization of N. gonorrhoeae by human PMNs. Using overlay assays and an affinity isolation technique, we demonstrate the direct interaction between Opa52-expressing gonococci and members of the human carcinoembryonic antigen (CEA) family which express the CD66 epitope. Gonococci and recombinant Escherichia coli strains synthesizing Opa52 showed specific binding and internalization by transfected HeLa cell lines expressing the CD66 family members BGP (CD66a), NCA (CD66c), CGM1 (CD66d) and CEA (CD66e), but not that expressing CGM6 (CD66b). Bacterial strains expressing either no opacity protein or the epithelial cell invasion-associated Opa50 do not bind these CEA family members. Consistent with their different receptor specificities, Opa52-mediated interactions could be inhibited by polyclonal anti-CEA sera, while Opa50 binding was instead inhibited by heparin. Using confocal laser scanning microscopy, we observed a marked recruitment of CD66 antigen by Opa52-expressing gonococci on both the transfected cell lines and infected PMNs. These data indicate that members of the CEA family constitute the cellular receptors for the interaction with, and internalization of, N. gonorrhoeae. PMID:9218786

  7. Low-Level Laser Therapy Attenuates LPS-Induced Rats Mastitis by Inhibiting Polymorphonuclear Neutrophil Adhesion

    PubMed Central

    WANG, Yueqiang; HE, Xianjing; HAO, Dandan; YU, Debin; LIANG, Jianbin; QU, Yanpeng; SUN, Dongbo; YANG, Bin; YANG, Keli; WU, Rui; WANG, Jianfa

    2014-01-01

    ABSTRACT The aim of this study was to investigate the effects of low-level laser therapy (LLLT) on a rat model of lipopolysaccharide (LPS)-induced mastitis and its underlying molecular mechanisms. The rat model of mastitis was induced by inoculation of LPS through the canals of the mammary gland. The results showed that LPS-induced secretion of IL-1? and IL-8 significantly decreased after LLLT (650 nm, 2.5 mW, 30 mW/cm2). LLLT also inhibited intercellular adhesion molecule-1 (ICAM-1) expression and attenuated the LPS-induced decrease of the expression of CD62L and increase of the expression of CD11b. Moreover, LLLT also suppressed LPS-induced polymorphonuclear neutrophils (PMNs) entering the alveoli of the mammary gland. The number of PMNs in the mammary alveolus and the myeloperoxidase (MPO) activity were decreased after LLLT. These results suggested that LLLT therapy is beneficial in decreasing the somatic cell count and improving milk nutritional quality in cows with an intramammary infection. PMID:25452258

  8. Complex dynamics of human red blood cell flickering: Alterations with in vivo aging Madalena Costa,1,* Ionita Ghiran,2,

    E-print Network

    cells RBCs , white blood cells leukocytes , and platelets. Newly formed "new" RBCs are released from in the liver and spleen. Thus, circulating RBCs represent a continuum of new to senescent "old" cells. As RBCs

  9. MEDIATORS OF INFLAMMATION IN LEUKOCYTE LYSOSOMES

    PubMed Central

    Janoff, Aaron; Schaefer, Sonja; Scherer, Joan; Bean, Michael A.

    1965-01-01

    The vascular permeability-increasing action of rabbit PMNL lysosomes has been studied in skin and cremaster muscle of the rat. Both an extract of frozen-thawed granules and a cathepsin-free cationic protein fraction of the granules (which had previously been demonstrated to cause leukocyte adhesion and emigration in vivo) induce increased vascular permeability in skin and muscle which resembles that produced by histamine or histamine-liberators with respect to the timing of the response and the predominant type of microvessel affected. Extracts of frozen-thawed lysosomes and the inflammatory lysosomal cationic protein both cause disruption of rat mesenteric mast cells in vitro, whereas a granule-free cytoplasmic fraction of PMN leukocytes and a non-inflammatory cationic protein fraction of the granules do not do so under identical test conditions. The mastocytolytic action of lysosomal materials in vitro is not inhibited in the presence of 10 kallikrein-inhibiting units of trasylol per ml. The mast cell rupturing fraction of PMNL granules (cationic protein) possesses no detectable peroxidase activity or acid-mucopolysaccharase activity. When compared with compound 48/80 on the basis of estimated molecular weight, the lysosomal cationic protein appears to be at least as active as the latter compound with respect to in vitro mastocytolytic potency. Chronic pretreatment of rats with an agent known to reduce tissue mast cell numbers causes marked suppression of the vascular permeability change normally induced in skin and muscle by lysosomal extracts and cationic protein. Similar results are obtained if lysosomal materials are tested in rats pretreated with an antihistaminic. These observations are discussed with respect to the mode of action of PMNL lysosomes in the early and late phases of local tissue-injury reactions. PMID:4379082

  10. Localization of indium-111 leukocytes in noninfected neoplasms

    SciTech Connect

    Lamki, L.M.; Kasi, L.P.; Haynie, T.P.

    1988-12-01

    Indium-111-labeled autologous leukocyte studies in general carry a high sensitivity, specificity, and accuracy for the investigation of infections and abscesses. However, past studies have described sporadic cases in which In leukocytes localized in tumors. Our experience using In leukocytes for the investigation of fever of unknown origin in cancer patients, however, indicates a relatively high incidence of In leukocyte localization in noninfected neoplasms. Out of the 61 patients studied for fever of unknown origin, 21 patients (34%) manifested abnormal localization of In leukocytes in neoplasms without clinical evidence of infection. These included patients with abnormal localization in: (a) lymph nodes, (b) soft-tissue tumors, and (c) bone neoplasms. The tumors included both primary and secondary lesions, and hematologic as well as solid tumors. The mechanism of In leukocyte localization in tumors is still not completely explained. Interpretations of In leukocyte studies in cancer patients with fever should take into consideration the possibility that localization may occur in neoplastic tissue per se and does not always indicate the presence of infection.

  11. Digital image analysis of blood cells.

    PubMed

    Da Costa, Lydie

    2015-03-01

    Rapid and accurate counts of red blood cells (RBCs), nucleated RBCs, platelets, and white blood cells (WBCs) (total and differential WBCs) are important requirements for a hematology laboratory. The detection of abnormal blood cell populations and the recognition of pathologic distributions of leukocytes are also of clinical importance. Manual microscopy counts are still required when a sample is flagged by the hematology analyzer and are still the reference method for WBC differential counts. Automated microscopy analyzers can provide accurate WBC differential counts, which may replace manual microscopy, but should not replace the eye of the cytologist. PMID:25676375

  12. Prepartal dietary energy level affects peripartal bovine blood neutrophil metabolic, antioxidant, and inflammatory gene expression.

    PubMed

    Zhou, Z; Bu, D P; Vailati Riboni, M; Khan, M J; Graugnard, D E; Luo, J; Cardoso, F C; Loor, J J

    2015-08-01

    During the dry period, cows can easily overconsume higher-grain diets, a scenario that could impair immune function during the peripartal period. Objectives were to investigate the effects of energy overfeeding on expression profile of genes associated with inflammation, lipid metabolism, and neutrophil function, in 12 multiparous Holstein cows (n=6/dietary group) fed control [CON, 1.34 Mcal/kg of dry matter (DM)] or higher-energy (HE, 1.62 Mcal/kg of DM) diets during the last 45 d of pregnancy. Blood was collected to evaluate 43 genes in polymorphonuclear neutrophil leukocytes (PMNL) isolated at -14, 7, and 14 d relative to parturition. We detected greater expression of inflammatory-related cytokines (IL1B, STAT3, NFKB1) and eicosanoid synthesis (ALOX5AP and PLA2G4A) in HE cows than in CON cows. Around parturition, all cows had a close balance in mRNA expression of the pro-inflammatory IL1B and the anti-inflammatory IL10, with greater expression of both in cows fed HE than CON. The expression of CCL2, LEPR, TLR4, IL6, and LTC4S was undetectable. Cows in the HE group had greater expression of genes involved in PMNL adhesion, motility, migration, and phagocytosis, which was similar to expression of genes related to the pro-inflammatory cytokine. This response suggests that HE cows experienced a chronic state of inflammation. The greater expression of G6PD in HE cows could have been associated with the greater plasma insulin, which would have diverted glucose to other tissues. Cows fed the HE diet also had greater expression of transcription factors involved in metabolism of long-chain fatty acids (PPARD, RXRA), suggesting that immune cells might be predisposed to use endogenous ligands such as nonesterified fatty acids available in the circulation when glucose is in high demand for milk synthesis. The lower overall expression of SLC2A1 postpartum than prepartum supports this suggestion. Targeting interleukin-1? signaling might be of value in terms of controlling the inflammatory response around calving. The present study revealed that overfeeding cows during late pregnancy results in activation, ahead of parturition, of PMNL responses associated with stress and inflammation. These adaptations observed in PMNL did not seem to be detrimental for production. PMID:26026766

  13. Imaging granularity of leukocytes with third harmonic generation microscopy

    PubMed Central

    Tsai, Cheng-Kun; Chen, Yu-Shing; Wu, Pei-Chun; Hsieh, Tsung-Yuan; Liu, Han-Wen; Yeh, Chiou-Yueh; Lin, Win-Li; Chia, Jean-San; Liu, Tzu-Ming

    2012-01-01

    Using third harmonic generation (THG) microscopy, we demonstrate that granularity differences of leukocytes can be revealed without a label. Excited by a 1230 nm femtosecond laser, THG signals were generated at a significantly higher level in neutrophils than other mononuclear cells, whereas signals in agranular lymphocytes were one order of magnitude smaller. Interestingly, the characteristic THG features can also be observed in vivo to track the newly recruited leukocytes following lipopolysaccharide (LPS) challenge. These results suggest that label-free THG imaging may provide timely tracking of leukocyte movement without disturbing the normal cellular or physiological status. PMID:23024916

  14. Direct viewing of atherosclerosis in vivo: plaque invasion by leukocytes is initiated by the endothelial selectins

    Microsoft Academic Search

    EINAR E. ERIKSSON; XUN XIE; JOACHIM WERR; PETER THOREN; LENNART LINDBOM

    2001-01-01

    Leukocyte infiltration in atherosclerosis has been extensively investigated by using histological techniques on fixed tissues. In this study, intravital microscopic observations of leukocyte recruitment in the aorta of atherosclerotic mice were performed. Interactions between leukocytes and atherosclerotic endothelium were highly transient, thereby limiting the ability for rolling leukocytes to firmly adhere. Leuko- cyte rolling was abolished by function inhibition of

  15. Microscale receiver operating characteristic analysis of micrometastasis recognition using activatable fluorescent probes indicates leukocyte imaging as a critical factor to enhance accuracy

    NASA Astrophysics Data System (ADS)

    Spring, Bryan Q.; Palanisami, Akilan; Hasan, Tayyaba

    2014-06-01

    Molecular-targeted probes are emerging with applications for optical biopsy of cancer. An underexplored potential clinical use of these probes is to monitor residual cancer micrometastases that escape cytoreductive surgery and chemotherapy. Here, we show that leukocytes, or white blood cells, residing in nontumor tissues-as well as those infiltrating micrometastatic lesions-uptake cancer cell-targeted, activatable immunoconjugates nonspecifically, which limits the accuracy and resolution of micrometastasis recognition using these probes. Receiver operating characteristic analysis of freshly excised tissues from a mouse model of peritoneal carcinomatosis suggests that dual-color imaging, adding an immunostain for leukocytes, offers promise for enabling accurate recognition of single cancer cells. Our results indicate that leukocyte identification improves micrometastasis recognition sensitivity and specificity from 92 to 93%-for multicellular metastases >20 to 30 ?m in size-to 98 to 99.9% for resolving metastases as small as a single cell.

  16. Methylation of leukocyte DNA and ovarian cancer: relationships with disease status and outcome

    PubMed Central

    2014-01-01

    Background Genome-wide interrogation of DNA methylation (DNAm) in blood-derived leukocytes has become feasible with the advent of CpG genotyping arrays. In epithelial ovarian cancer (EOC), one report found substantial DNAm differences between cases and controls; however, many of these disease-associated CpGs were attributed to differences in white blood cell type distributions. Methods We examined blood-based DNAm in 336 EOC cases and 398 controls; we included only high-quality CpG loci that did not show evidence of association with white blood cell type distributions to evaluate association with case status and overall survival. Results Of 13,816 CpGs, no significant associations were observed with survival, although eight CpGs associated with survival at p?leukocytes that are differentially methylated by case-control status. Since a retrospective study design was used, we cannot differentiate whether DNAm was etiologic or resulting from EOC; thus, prospective studies of EOC-associated loci are the critical next step. PMID:24774302

  17. Quantifying transient psychological stress using a novel technique: changes to PMA-induced leukocyte production of ROS in vitro.

    PubMed

    Shelton-Rayner, Graham K; Mian, Rubina; Chandler, Simon; Robertson, Duncan; Macdonald, David W

    2011-01-01

    Although much work has been conducted to quantify the long-term physiological effects of psychological stress, measures of short-term, low-level stress have been more elusive. This study assessed the effect of exposure of volunteers to a mild, brief, psychologically stressful event, on the functional ability of leukocytes in whole blood to respond to phorbol 12-myristate 13-acetate (PMA) in vitro. Volunteers operated a car electric window and adjusted it to 4 pre-determined positions. Between each operation the mechanism's polarity was covertly altered (group B) or remained unaltered (group A). For each treatment group 10 different subjects provided capillary blood samples pre- and post-stressor. Using a chemiluminescent technique termed leukocyte coping capacity, the ability of leukocytes to produce reactive oxygen species (ROS) in vitro was assessed. ROS release differed significantly at 10 min post-stressor between treatment groups, suggesting exposure to acute psychological stress leads to a reduced ability to respond to bacterial challenge. PMID:21375949

  18. Tumor Cell Extravasation Mediated by Leukocyte Adhesion is Shear Rate Dependent on IL-8 Signaling*

    PubMed Central

    Liang, Shile; Hoskins, Meghan; Dong, Cheng

    2009-01-01

    To complete the metastatic journey, cancer cells have to disseminate through the circulation and extravasate to distal organs. However, the extravasation process, by which tumor cells leave a blood vessel and invade the surrounding tissue from the microcirculation, remains poorly understood at the molecular level. In this study, tumor cell adhesion to the endothelium (EC) and subsequent extravasation were investigated under various flow conditions. Results have shown polymorphonuclear neutrophils (PMNs) facilitate melanoma cell adhesion to the EC and subsequent extravasation by a shear-rate dependent mechanism. Melanoma cell-PMN interactions are mediated by the binding between intercellular adhesion molecule-1 (ICAM-1) on melanoma cells and ?2 integrins on PMNs. In addition, the fluid convection affects the extent of activation of ?2 integrins on PMNs by endogenously secreted interleukin 8 (IL-8) within the tumor microenvironment. Results also indicate that shear rate affects the binding kinetics between PMNs and melanoma cells, which may contribute to the shear-rate dependence of melanoma extravasation in a shear flow when mediated by PMNs. PMID:20379392

  19. Cord Blood Transplantation: Can We Make it Better?

    PubMed Central

    Metheny, Leland; Caimi, Paolo; de Lima, Marcos

    2013-01-01

    Umbilical cord blood is an established source of hematopoietic stem cells for transplantation. It enjoys several advantages over bone marrow or peripheral blood, including increased tolerance for Human Leukocyte Antigen mismatches, decreased incidence of graft-versus-host disease, and easy availability. Unrelated cord blood does have limitations, however, especially in the treatment of adults. In the 24?years since the first umbilical cord blood transplant was performed, significant progress has been made, but delayed hematopoietic engraftment and increased treatment-related mortality remain obstacles to widespread use. Here we summarize the latest results of unrelated cord blood transplants, and review strategies under investigation to improve clinical outcomes. PMID:24062989

  20. Blood culture

    MedlinePLUS

    Culture - blood ... A blood sample is needed. The site where blood will be drawn is first cleaned with an antiseptic such ... organism from the skin getting into (contaminating) the blood sample and causing a false-positive result (see ...

  1. Blood Transfusion

    MedlinePLUS

    ... from the NHLBI on Twitter. What Is a Blood Transfusion? A blood transfusion is a safe, common ... Very rarely, serious problems develop. Important Information About Blood The heart pumps blood through a network of ...

  2. Blood clotting

    MedlinePLUS Videos and Cool Tools

    ... the external bleeding stops. Clotting factors in the blood cause strands of blood-borne material, called fibrin, to stick together and ... the inside of the wound. Eventually, the cut blood vessel heals, and the blood clot dissolves after ...

  3. What's Blood?

    MedlinePLUS

    ... Let's find out more about each ingredient. Continue Red Blood Cells Red blood cells (also called erythrocytes, say: ih- rith - ... Most of the cells in the blood are red blood cells. They carry around an important chemical ...

  4. ADRENERGIC NERVES GOVERN CIRCADIAN LEUKOCYTE RECRUITMENT TO TISSUES

    PubMed Central

    Scheiermann, Christoph; Kunisaki, Yuya; Lucas, Daniel; Chow, Andrew; Jang, Jung-Eun; Zhang, Dachuan; Hashimoto, Daigo; Merad, Miriam; Frenette, Paul S.

    2012-01-01

    SUMMARY The multistep sequence leading to leukocyte migration is thought to be locally regulated at the inflammatory site. Here, we show that broad systemic programs involving long-range signals from the sympathetic nervous system (SNS) delivered by adrenergic nerves, regulate rhythmic recruitment of leukocytes in tissues. Constitutive leukocyte adhesion and migration in murine bone marrow (BM) and skeletal muscle microvasculature fluctuated with circadian peak values at night. Migratory oscillations, altered by experimental jetlag, were implemented by perivascular SNS fibers, acting on ?-adrenoreceptors expressed on non-hematopoietic cells, and leading to tissue-specific, differential circadian oscillations in the expression of endothelial cell adhesion molecules and chemokines. We showed that these rhythms have physiological consequences by alteration of hematopoietic cell recruitment and overall survival in models of septic shock, sickle cell vaso-occlusion and BM transplantation. These data provide unique insight in the leukocyte adhesion cascade and the potential for time-based therapeutics for transplantation and inflammatory diseases. PMID:22863835

  5. 21 CFR 864.7675 - Leukocyte peroxidase test.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7675 Leukocyte peroxidase test. (a) Identification. A...

  6. 21 CFR 864.7660 - Leukocyte alkaline phosphatase test.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7660 Leukocyte alkaline phosphatase test. (a)...

  7. 21 CFR 864.7675 - Leukocyte peroxidase test.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7675 Leukocyte peroxidase test. (a) Identification. A...

  8. 21 CFR 864.7660 - Leukocyte alkaline phosphatase test.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7660 Leukocyte alkaline phosphatase test. (a)...

  9. 21 CFR 864.7660 - Leukocyte alkaline phosphatase test.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7660 Leukocyte alkaline phosphatase test. (a)...

  10. 21 CFR 864.7660 - Leukocyte alkaline phosphatase test.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7660 Leukocyte alkaline phosphatase test. (a)...

  11. 21 CFR 864.7675 - Leukocyte peroxidase test.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7675 Leukocyte peroxidase test. (a) Identification. A...

  12. Influence of Magnetite Nanoparticles on Human Leukocyte Activity

    NASA Astrophysics Data System (ADS)

    Džarová, Anežka; Dubni?ková, Martina; Závišová, Vlasta; Koneracká, Martina; Kop?anský, Peter; Gojzewski, Hubert; Timko, Milan

    2010-12-01

    Chemically synthesized magnetite particles coated by sodium oleate and PEG (MNP), and magnetosomes (MS) influence the process of phagocytosis and the metabolic activity (lysozyme and peroxidase activity) in leukocytes. Lysozyme activity is oxygen-independent liquidation mechanisms of engulfed microorganism, peroxidase activity is an oxygen-dependent mechanism. Both tested types of nanoparticles lysed leukocyte cells during incubation. MNP at concentrations of 10 and 20 ?g/mL lysed almost all leukocytes and their cell viability was in the 14±0.05% range. On the other hand MS begin to influence leukocytes activity at the concentration of 1 ?g/ml and this influence grows with increasing concentration up to 20 ?g/ml. MS are more suitable for biological applications than MNP which are more aggressive material than MS. MS should not be used above 10 ?g/mL.

  13. Osteomyelitis complicating fracture: pitfalls of /sup 111/In leukocyte scintigraphy

    SciTech Connect

    Kim, E.E.; Pjura, G.A.; Lowry, P.A.; Gobuty, A.H.; Traina, J.F.

    1987-05-01

    /sup 111/In-labeled leukocyte imaging has shown greater accuracy and specificity than alternative noninvasive methods in the detection of uncomplicated osteomyelitis. Forty patients with suspected osteomyelitis complicating fractures (with and without surgical intervention) were evaluated with /sup 111/In-labeled leukocytes. All five patients with intense focal uptake, but only one of 13 with no uptake, had active osteomyelitis. However, mild to moderate /sup 111/In leukocyte uptake, observed in 22 cases, indicated the presence of osteomyelitis in only four of these; the other false-positive results were observed in noninfected callus formation, heterotopic bone formation, myositis ossificans, and sickle-cell disease. These results suggest that /sup 111/In-labeled leukocyte imaging is useful for the evaluation of suspected osteomyelitis complicating fracture but must be used in conjunction with clinical and radiographic correlation to avoid false-positive results.

  14. Rehabilitation of ischemically damaged human kidneys by normothermic extracorporal hemoperfusion in situ with oxygenation and leukocyte depletion.

    PubMed

    Reznik, O; Bagnenko, S; Skvortsov, A; Ananyev, A; Senchik, K; Loginov, I; Moysyuk, Y

    2010-06-01

    The potential use of donors after cardiac death is an important issue. The crucial point in uncontrolled donors after cardiac death is the warm ischemic time. Definition of the limit of warm ischemic time was 1 of our tasks; an other was to develop a preservation protocol. Five uncontrolled donors kidneys with warm ischemia times up to 80 minutes were exposed to an extracorporal perfusion device for preservation after ischemic damage. We sought to eliminate leukocytes as the main damaging factor in modified donor blood circulating in the device. Recipients of 8 kidneys from this protocol displayed immediate graft function with an average serum creatinine value of 140 +/- 37.9 mmol/L at the end of the second month. Treatment of ischemically damaged kidneys by normothermic extracorporal perfusion with leukocyte depletion before procurement demands further study. PMID:20620470

  15. Isolation of leukocytes from the human maternal-fetal interface

    PubMed Central

    Xu, Yi; Plazyo, Olesya; Romero, Roberto; Hassan, Sonia S.; Gomez-Lopez, Nardhy

    2015-01-01

    Pregnancy resembles an inflammatory response that is characterized by the infiltration of leukocytes in the reproductive tissues and at the maternal-fetal interface (decidua basalis and decidua parietalis). This interface is the anatomical site of contact between maternal and fetal tissues; therefore, it is an immunological site of action during pregnancy. Infiltrating leukocytes at the maternal-fetal interface play a central role in implantation, pregnancy maintenance, and timing of delivery. Therefore, phenotypic and functional characterizations of these leukocytes will provide insight into the pathogenesis of pregnancy-related complications. Several protocols have been described in order to isolate infiltrating leukocytes from the decidua basalis and decidua parietalis; however, the lack of consistency in the reagents, enzymes, and times of incubation makes it difficult to compare these results. Herein is described a novel approach that combines the use of gentle mechanical and enzymatic dissociation techniques to preserve the viability and integrity of extracellular and intracellular markers in leukocytes isolated from the human tissues at the maternal-fetal interface. Aside from immunophenotyping, cell culture and sorting, the future applications of this protocol are numerous and varied. Following this protocol, the isolated leukocytes can be used to determine DNA methylation, expression of target genes, in vitro leukocyte functionality (i.e., phagocytosis, cytotoxicity, T-cell proliferation, and plasticity, etc.), and the production of reactive oxygen species at the maternal-fetal interface. Additionally, using the described protocol, this laboratory has been able to describe new and rare leukocytes at the maternal-fetal interface. PMID:26067211

  16. Rose hip inhibits chemotaxis and chemiluminescence of human peripheral blood neutrophils in vitro and reduces certain inflammatory parameters in vivo

    Microsoft Academic Search

    Arsalan Kharazmi; Kaj Winther

    1999-01-01

    Objective and Design  The objective of this study was to investigate the leucocyte-related antiinflammatory properties of rose hip. Materials and\\u000a Methods: The effect of rose hip on a number of inflammatory parameters was evaluated using the following models: (1) The effect\\u000a of rose hip extract on chemotaxis and chemiluminescence of peripheral blood polymorphonuclear leucocytes (PMNs) from healthy\\u000a subjects in vitro; (2)

  17. Lessons from rare maladies: leukocyte adhesion deficiency syndromes

    PubMed Central

    Harris, Estelle S.; Weyrich, Andrew S.; Zimmerman, Guy A.

    2013-01-01

    Purpose of review The leukocyte adhesion deficiency (LAD) syndromes are rare genetically determined conditions with challenging clinical features. These immunodeficiencies also provide insights that are broadly relevant to the biology of leukocytes, platelets, intercellular interactions, and intracellular signaling. Recent discoveries merit their review in the context of existing knowledge. Recent findings New activities of ?2 integrins, which are deficient or absent in LAD-I, and new ?2 integrin-dependent functions of neutrophils and other leukocytes have recently been identified. Genetic defects and mechanisms accounting for impaired fucosylation of selectin ligands and defective selectin binding and signaling in LAD-II are now apparent. LAD-III, which presents with bleeding similar to that in Glanzmann thrombasthenia and platelet dysfunction in addition to impaired leukocyte adhesion, is now known to be due to absence of KINDLIN-3, a cytoplasmic protein that acts cooperatively with TALIN-1 in activating ?1, ?2, and ?3 integrins. Understanding of the leukocyte adhesion cascade and interactions of leukocytes with inflamed endothelium, which are impaired in each of the LAD syndromes, continues to be refined. Summary Although LAD syndromes are rare maladies, their investigation is generating new knowledge directly applicable to the diagnosis and care of patients and to fundamental paradigms in immunobiology and hemostasis. PMID:23207660

  18. Constitutive and agonist stimulated ATP secretion in leukocytes

    PubMed Central

    Campwala, Hinnah; Fountain, Samuel J.

    2013-01-01

    Release and reception of extracellular ATP by leukocytes plays a critical role in immune responses to infection, injury and cardiovascular disease. Leukocytes of both the innate, adaptive immune and central nervous system express a repertoire of cell surface receptors for ATP (P2X and P2Y receptors) and its metabolites. ATP acts as a damage-associated molecule pattern (DAMP) released by injured or dying cells. Detection of released ATP by neighboring leukocytes initiates inflammation and wound healing. However, recent evidence from our group and others suggests ATP release by leukocytes themselves serves to regulate homeostatic mechanisms and coordinate responses to external pro-inflammatory cues. Examples include the homeostatic control of intracellular calcium and regulation of migratory guidance during chemotactic response to external cues. Though there has been some progress in elucidating ATP release mechanisms of some mammalian cells types, release conduits and coupling signal transduction machinery remain larger elusive for leukocytes. Our recent studies suggest a role for secretory lysosomes in releasing ATP in monocytes. Though poorly defined, targeting ATP release mechanisms in leukocytes have great anti-inflammatory potential. PMID:23713132

  19. Physiological levels of testosterone kill salmonid leukocytes in vitro

    USGS Publications Warehouse

    Slater, C.H.; Schreck, C.B.

    1997-01-01

    Adult spring chinook salmon (Oncorhynchus tshawytscha) elaborate high plasma concentrations of testosterone during sexual maturation, and these levels of testosterone have been shown to reduce the salmonid immune response in vitro. Our search for the mechanism of testosterone's immunosuppressive action has led to the characterization of an androgen receptor in salmonid leukocytes. In the present study we examined the specific effects that testosterone had on salmonid leukocytes. Direct counts of viable leukocytes after incubation with and without physiological levels of testosterone demonstrate a significant loss of leukocytes in cultures exposed to testosterone. At least 5 days of contact with testosterone was required to produce significant immunosuppression and addition of a 'conditioned media' (supernatant from proliferating lymphocytes not exposed to testosterone) did not reverse the immunosuppressive effects of testosterone. These data lead us to conclude that testosterone may exert its immunosuppressive effects by direct action on salmonid leukocytes, through the androgen receptor described, and that this action leads to the death of a significant number of these leukocytes.

  20. A Monte Carlo approach to rolling leukocyte tracking in vivo.

    PubMed

    Cui, Jing; Acton, Scott T; Lin, Zongli

    2006-08-01

    Tracking the movement of rolling leukocytes in vivo contributes to the understanding of the mechanism of the inflammatory process and to the development of anti-inflammatory drugs. Several roadblocks exist that hinder successful automated tracking including the moving background, the severe image noise and clutter, the occlusion of the target leukocyte by other leukocytes and structures, the jitter caused by the breathing movement of the living animal, and the weak image contrast. In this paper, a Monte Carlo tracker is developed for automatically tracking a single rolling leukocyte in vivo. Based on the leukocyte movement information and the image intensity features, a specialized sample-weighting criterion is tailored to the application. In comparison with a snake-based tracker, our experiments show that, as the noise intensity level increases, the performance of the snake tracker degrades more than that of the Monte Carlo tracker. In cases, where the leukocyte is observed in contact with the vessel wall, the Monte Carlo tracker is less affected by the image clutter. From tracking within 99 intravital microscopic video sequences, the Monte Carlo tracker exhibits superior performance in the reduced localization error and the increased number of frames tracked when compared with the centroid tracker, the correlation tracker and the GVF snake tracker. PMID:16876461