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1

A simple method for measuring the F-actin content of human polymorphonuclear leukocytes in whole blood  

Microsoft Academic Search

We have developed an improved method for measuring the filamentous (F) actin content of human blood polymorphonuclear leukocytes (PMNs). The essential feature of the method is the immediate fixation of the F-actin cytoskeleton. Fresh whole blood (100 µl) is shock-cooled by the addition of 1.0 ml of a mixture of 18.75% glycerol and 5% formaldehyde in phosphate buffer pre-cooled to

Gerd Egger; Astrid Burda; Andreas Glasner

2001-01-01

2

Poly(ethylene glycol)-containing hydrogels promote the release of primary granules from human blood-derived polymorphonuclear leukocytes.  

PubMed

Polymorphonuclear leukocytes (PMNs) are recruited to sites of injury and biomaterial implants. Once activated, PMNs can exocytose their granule subsets to recruit monocytes (MCs) and mediate MC/macrophage activation. We investigated the release of myeloperoxidase (MPO), a primary granule marker, and matrix metalloproteinase-9 (MMP-9), a tertiary granule marker, from human blood-derived PMNs cultured on poly(ethylene glycol) (PEG) hydrogels, polydimethylsiloxane (PDMS), tissue culture polystyrene (TCPS) and gelatin-PEG (GP) hydrogels, with and without the presence of the bacterial peptide formyl-Met-Leu-Phe. Supernatants from PMN cultures on PEG-containing hydrogels (i.e., PEG and GP hydrogels) had higher concentrations of MPO than those from PMN cultures on PDMS or TCPS at 2 h. PMNs on all biomaterials released comparable levels of MMP-9 at 2 h, indicating that PMNs cultured on PEG-containing hydrogels have different mechanisms of release for primary and tertiary granules. Src family kinases were involved in the release of MPO from PMNs cultured on PEG hydrogels, TCPS and GP hydrogels and in the release of MMP-9 from PMNs cultured on all four biomaterials. The increased release of primary granules from PMNs on PEG-containing hydrogels did not significantly increase MC chemotaxis, indicating that additional co-effectors in the dynamic inflammatory milieu in vivo modulate PMN-mediated MC recruitment. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 4252-4261, 2014. PMID:24497370

Cohen, Hannah Caitlin; Lieberthal, Tyler Jacob; Kao, W John

2014-12-01

3

Oxidative DNA damage of peripheral blood polymorphonuclear leukocytes, selectively induced by chronic arsenic exposure, is associated with extent of arsenic-related skin lesions  

SciTech Connect

There is increasing evidence that oxidative stress is an important risk factor for arsenic-related diseases. Peripheral blood leukocytes constitute an important defense against microorganisms or pathogens, while the research on the impact of chronic arsenic exposure on peripheral blood leukocytes is much more limited, especially at low level arsenic exposure. The purpose of the present study was to explore whether chronic arsenic exposure affects oxidative stress of peripheral blood leukocytes and possible linkages between oxidative stress and arsenic-induced skin lesions. 75 male inhabitants recruited from an As-endemic region of China were investigated in the present study. The classification of arsenicosis was based on the degree of skin lesions. Arsenic levels were measured in drinking water and urine by Atomic Fluorescence Spectroscopy. Urinary 8-hydroxy-2?-deoxyguanosine (8-OHdG) was tested by Enzyme-Linked Immunosorbent Assay. 8-OHdG of peripheral blood leukocytes was evaluated using immunocytochemical staining. 8-OHdG-positive reactions were only present in polymorphonuclear leukocytes (PMNs), but not in monocytes (MNs). The 8-OHdG staining of PMN cytoplasm was observed in all investigated populations, while the 8-OHdG staining of PMN nuclei was frequently found along with the elevated amounts of cell debris in individuals with skin lesion. Urinary arsenic levels were increased in the severe skin lesion group compared with the normal group. No relationship was observed between drinking water arsenic or urine 8-OHdG and the degree of skin lesions. These findings indicated that the target and persistent oxidative stress in peripheral blood PMNs may be employed as a sensitive biomarker directly to assess adverse health effects caused by chronic exposure to lower levels of arsenic. -- Highlights: ? Male inhabitants were investigated from an As-endemic region of China. ? 8-OHdG-positive reactions were only present in polymorphonuclear leukocytes (PMNs). ? 8-OHdG staining of PMN nuclei was paralleled by increased debris of cells. ? Oxidative DNA damage of PMNs is associated with arsenic-related skin lesions.

Pei, Qiuling, E-mail: 924969007@qq.com [Department of Toxicology, Public Health College, Shanxi Medical University, No 56 Xin Jian Nan Lu, Taiyuan (030001) (China)] [Department of Toxicology, Public Health College, Shanxi Medical University, No 56 Xin Jian Nan Lu, Taiyuan (030001) (China); Ma, Ning [Faculty of Health Science, Suzuka University of Medical Science, Suzuka, 510-0293 (Japan)] [Faculty of Health Science, Suzuka University of Medical Science, Suzuka, 510-0293 (Japan); Zhang, Jing; Xu, Wenchao; Li, Yong; Ma, Zhifeng; Li, Yunyun; Tian, Fengjie; Zhang, Wenping [Department of Toxicology, Public Health College, Shanxi Medical University, No 56 Xin Jian Nan Lu, Taiyuan (030001) (China)] [Department of Toxicology, Public Health College, Shanxi Medical University, No 56 Xin Jian Nan Lu, Taiyuan (030001) (China); Mu, Jinjun [The Second Hospital, Shanxi Medical University, Taiyuan (030001) (China)] [The Second Hospital, Shanxi Medical University, Taiyuan (030001) (China); Li, Yuanfei [The First Hospital, Shanxi Medical University, Taiyuan (030001) (China)] [The First Hospital, Shanxi Medical University, Taiyuan (030001) (China); Wang, Dongxing; Liu, Haifang; Yang, Mimi; Ma, Caifeng; Yun, Fen [Department of Toxicology, Public Health College, Shanxi Medical University, No 56 Xin Jian Nan Lu, Taiyuan (030001) (China)] [Department of Toxicology, Public Health College, Shanxi Medical University, No 56 Xin Jian Nan Lu, Taiyuan (030001) (China)

2013-01-01

4

Functional activity of enucleated human polymorphonuclear leukocytes  

PubMed Central

Enucleated human polymorphonuclear leukocytes (PMN) were prepared by centrifuging isolated, intact PMN over a discontinuous Ficoll gradient that contained 20 microM cytochalasin B. The enucleated cells (PMN cytoplasts) contained about one-third of the plasma membrane and about one-half of the cytoplasm present in intact PMN. The PMN cytoplasts contained no nucleus and hardly any granules. The volume of the PMN cytoplasts was about one-fourth of that of the original PMN. Greater than 90% of the PMN cytoplasts had an "outside-out" topography of the plasma membrane. Cytoplasts prepared from resting PMN did not generate superoxide radicals (O2-) or hydrogen peroxide. PMN cytoplasts incubated with opsonized zymosan particles or phorbol-myristate acetate induced a respiratory burst that was qualitatively (O2 consumption, O2- and H2O2 generation) and quantitatively (per unit area of plasma membrane) comparable with that of intact, stimulated PMN. Moreover, at low ratios of bacteria/cells, PMN cytoplasts ingested opsonized Staphylococcus aureus bacteria as well as did intact PMN. At higher ratios, the cytoplasts phagocytosed less well. The killing of these bacteria by PMN cytoplasts was slower than by intact cells. The chemotactic activity of PMN cytoplasts was very low. These results indicate that the PMN apparatus for phagocytosis, generation of bactericidal oxygen compounds, and killing of bacteria, as well as the mechanism for recognizing opsonins and activating PMN functions, are present in the plasma membrane and cytosol of these cells. PMID:6309859

1983-01-01

5

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

NASA Technical Reports Server (NTRS)

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.

Martin, R. R.

1976-01-01

6

Promoting Effect of Colostrum on the Phagocytic Activity of Bovine Polymorphonuclear Leukocytes in vitro  

Microsoft Academic Search

Bovine colostrum contains a variety of essential nutrients, antibodies, cytokines, hormones, and growth factors that are important for nutrient supply, host defense, growth and for general neonatal adaptation. We have investigated the effect of bovine colostrum on the phagocytic activity for latex particles by normal peripheral blood polymorphonuclear leukocytes using flow cytometric analysis. The phagocytosis promoting effect was observed in

H. Sugisawa; T. Itou; T. Sakai

2001-01-01

7

Isolation, In-111 labeling, and abscess detection efficiency of rabbit polymorphonuclear leukocytes (PMN) from blood and peritoneal fluid  

SciTech Connect

In-111 labeled blood and peritoneal exudate PMN were compared for labeling efficiency and ability to migrate to sites of experimental abscesses using both direct sampling and visual imaging techniques. Blood PMN were prepared by combining heparinized blood with 6% Hetastarch for 1 hour and layering the plasma over a double density Ficoll-Hy-paque gradient (S.G. 1.076 over 1.141). The PMN layer (90-99% PMN) at the interface yielded 10/sup 6/-10/sup 7/ PMN from 80-120 ml of blood. Peritoneal PMN were obtained by infusion of 0.1% glycogen, followed by infusion of saline after 4 or 18 hours. The exudate yielded 10/sup 7/-10/sup 8/ PMN (80-99% PMN). PMN suspensions were labeled for 30 minutes by addition of 100 ..mu..Ci of In-111-oxine, then washed twice. Percent cell-associated radioactivity of the labeled blood, 4 hour, and 18 hour peritoneal PMN was 89%, 88%, and 86%. The labeled PMN were injected intravenously into rabbits which had two of three abdominal capsules (table tennis balls drilled with 250 1.5 mm holes) inoculated with Staphylococcus aureus 4 hours earlier. Peak venous recovery of circulating labeled PMN, for blood, 4 hour and 18 hour peritoneal PMN was 60%, 43%, and 19%. Gamma camera images 24 hours after infusion into infected rabbits were superior with 4 hour peritoneal PMN. The peritoneal PMN harvested 4 hours after glycogen stimulation are simple to prepare, are obtainable in greater numbers than blood PMN, and result in better abscess visualization.

Bettin, K.M.; Elson, M.K.; Gerding, D.N.; Bamberger, D.M.; Forstrom, L.A.; Shafer, R.B.

1984-01-01

8

Exocytosis of polymorphonuclear leukocyte lysosomal contents induced by dental plaque.  

PubMed Central

Rabbit polymorphonuclear leukocytes were incubated with a sonically treated suspension of pooled dental plaque to determine if the plaque would induce release of lysosomal enzymes from the polymorphonuclear leukocytes. Cells incubated with plaque at 37 degrees C released significantly greater amounts of the lysosomal enzymes, beta-glucuronidase and lysozyme, than did cells incubated with plaque at 0 degrees C or without plaque at 37 degrees C. This response was both dose and time dependent. Release of the cytoplasmic enzyme lactate dehydrogenase was minimal, and there were no significant differences in lactate dehydrogenase release between cells at 0 and 37 degrees C, or without plaque. These results indicate that dental plaque can induce the selective release of lysosomal enzymes, which could be involved in the periodontal injury produced by dental plaque. PMID:561032

White, R R; Montgomery, E H

1977-01-01

9

Effects of lead on the killing mechanisms of polymorphonuclear leukocytes  

SciTech Connect

The effects of lead on the killing mechanisms of rat polymorphonuclear leukocytes (PMN) were investigated, using male Long-Evans rats exposed to 1% lead acetate in the drinking water for varying periods of time to achieve blood lead levels ranging from 20-200 ..mu..g/dl. Studies of PMN bacterial and fungal killing activity, chemotaxis and phagocytosis demonstrated that: 1) bactericidal activity of PMN from rats exposed to lead was not altered; 2) chemotactic activity remained within normal limits; 3) the phagocytic ability of the PMN also remained unaltered. In addition to these normal findings, one major abnormality was demonstrated: a significant decrease in the ability of PMN from rats exposed to lead to kill Candida albicans. This defect was not related to age or to length of exposure. It could not be produced by addition of lead to the test system in vitro. Further investigation revealed significant decreases in PMN glucose-6-phosphate dehydrogenase, catalase, and myeloperoxidase activities. These data support two possible mechanisms for the abnormal fungicidal activity of PMN from lead-exposed rats: decrease in ability to reduce oxygen to active metabolites, or reduction in myeloperoxidase activity due to diminshed synthesis of the heme moiety required for its function.

Silberstein, C.F.

1984-01-01

10

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

Microsoft Academic Search

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

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

2003-01-01

11

Effect of Nedocromil Sodium on Polymorphonuclear Leukocyte Plasma Membrane  

PubMed Central

The effect of nedocromil sodium on the plasma membrane fluidity of polymorphonuclear leukocytes (PMNs) was investigated by measuring steady-state fluorescence anisotropy of 1-[4-trimethylammonium-phenyl]-6-phenyl- 1,3,5-hexatriene (TMA-DPH) incorporated in the membrane. Our results show that nedocromil sodium 300 ?M significantly decreased membrane fluidity of PMNs. The decrease in membrane fluidity of PMNs induced by fMLP was abolished in the presence of nedocromil sodium. These data suggest that nedocromil sodium interferes with the plasma membranes of PMNs and modulates their activities. PMID:18475598

Oggiano, N.; Giorgi, P. L.; Coppa, G. V.; Gabbianelli, R.; Bruni, S.; Cutrona, F. M.; Fiorini, R.

1994-01-01

12

Influenza A virus-induced polymorphonuclear leukocyte dysfunction in the pathogenesis of experimental pneumococcal otitis media.  

PubMed Central

The role of influenza A virus-induced polymorphonuclear leukocyte and eustachian tube dysfunction in the pathogenesis of acute purulent otitis media was studied in chinchillas. Polymorphonuclear leukocyte function, middle ear pressure, and the incidence of pneumococcal otitis media were observed after intranasal inoculation with influenza A virus, Streptococcus pneumoniae, or both. Results showed that depressed negative middle ear pressure and polymorphonuclear leukocyte chemiluminescence and chemotactic activity occurred after influenza inoculation, but not after inoculation with pneumococcus alone. The greatest incidence of pneumococcal otitis media occurred when the pneumococcus was inoculated just before the time of influenza-induced polymorphonuclear leukocyte dysfunction and negative middle ear pressure. Animals that had unilateral tympanostomy tubes placed before inoculation of influenza with pneumococcus showed no difference in the occurrence of pneumococcal otitis media in ventilated and nonventilated ears, suggesting that polymorphonuclear leukocyte dysfunction contributes more to the pathogenesis of pneumococcal otitis media than does negative middle ear pressure in this animal model. PMID:7076299

Abramson, J S; Giebink, G S; Quie, P G

1982-01-01

13

[The chemiluminescence of the polymorphonuclear leukocytes in food poisoning].  

PubMed

Luminol- and lucigenin-dependent chemiluminescence (CL) induced by zymosan, opsonized autoserum was used to study the oxygen-dependent bactericidal system of polymorphonuclear leukocytes (PNL) in patients with food toxico-infections, at different disease periods. Different modifications of CL were established to have analogous dynamics on days 1 and 2 of the disease and differences as to the period of convalescence. A correlation was revealed between the amplitude of CL and the intensity of the intoxication syndrome. The conclusion is made about the role played by granulocytes in the disease pathogenesis. The authors review potential mechanisms of participation of PNL free radicals in the pathophysiological shifts responsible for the development, course and outcome of food toxico-infections. PMID:2094985

Popov, P Iu; Rosly?, I M; Malov, V A; Pak, S G

1990-01-01

14

Evidence for polymorphonuclear leukocyte collagenase and 92-kilodalton gelatinase in gingival crevicular fluid.  

PubMed Central

Analysis of inflammatory exudate collected from sites of experimental periodontitis in cynomolgus monkeys has revealed the presence of collagenase and a 92-kDa gelatinase that comigrated after electrophoresis with the 92-kDa gelatinase released from polymorphonuclear leukocytes. Since neutralizing antibodies to fibroblast collagenase had no effect on the collagenase activity and bacterial collagenases could not be detected, polymorphonuclear leukocytes appear to be the major source of collagenolytic proteinases in inflammatory fluid from gingiva. Images PMID:1657787

Overall, C M; Sodek, J; McCulloch, C A; Birek, P

1991-01-01

15

Physiologic-Chemoattractant-Induced Migration of Polymorphonuclear Leukocytes in Milk  

PubMed Central

The somatic cell count (SCC; leukocytes and epithelial cells) in milk is used as an indicator of udder health status. A SCC above the regulatory standard is generally considered as an indication of mastitis. Therefore, milk with a SCC equal to or greater than the regulatory limit cannot be sold to the public because it is unsuitable for human consumption. This study was performed to determine whether SCC levels above the regulatory limit observed in goats during late lactation are a physiologic or a pathological response of the goat mammary gland. Differential counts of cells in nonmastitic goat milk samples during late lactation revealed that approximately 80% of the cells were polymorphonuclear leukocytes (PMNs). In addition, microchemotaxis assay results indicated that normal nonmastitic late-lactation-stage goat milk is significantly higher (P < 0.001) in PMN chemotactic activity than early-lactation-stage goat milk, with a mean chemotactic activity of 14.9 and 42.7/mg of protein for early and late lactation stages, respectively. Physicochemical analyses also suggest that the PMN infiltration observed in normal late-lactation-stage goat milk is due to a PMN chemotactic factor(s) that is different from the PMN chemotactic factor(s) present in mastitic milk. Interestingly, the PMN chemotactic factor in late-lactation-stage goat milk is highly acid resistant (pH 2), suggesting that the factor is able to survive the highly acidic gastric environment and may therefore be important in the augmentation of the immune systems of sucklings. These results indicate that the chemotactic factor(s) present in the milk of normal late-lactation-stage goats is nonpathological and may play a physiologic regulatory role in mammary gland involution. Hence, the regulatory standard for goat milk needs to be redefined in order to reflect this. PMID:9605994

Manlongat, Natasha; Yang, T. J.; Hinckley, L. S.; Bendel, Robert B.; Krider, H. M.

1998-01-01

16

Metabolism of 2-aminofluorene by human polymorphonuclear leukocytes: more evidence for the association between inflammation and cancer.  

PubMed Central

Recent investigations have demonstrated the ability of leukocytes to metabolize promutagens or procarcinogens into their genotoxic forms. As a possible explanation for the association between inflammation and cancer, we and others have hypothesized that local accumulations of leukocytes could take up nearby promutagens, metabolize them, and release genotoxic agents that may cause damage in the surrounding tissue. Using a modified, two-step preincubation protocol with Salmonella, we have tested this hypothesis. We have shown that total human peripheral blood leukocytes, cultured in the presence of 2-aminofluorene for 18 hr, can metabolize 2-aminofluorene into agents mutagenic to Salmonella typhimurium strain TA98. Furthermore, experiments in which polymorphonuclear leukocytes were separated from mononuclear leukocytes demonstrated that the PMNs metabolized 2-aminofluorene to a much greater extent than the MNs. PMID:8143630

Isola, V J; Hartman, T C; Trumble, S J; Ruzek, M C; Gentile, J M

1993-01-01

17

Extravasation of polymorphonuclear leukocytes from the cerebral microvasculature. Inflammatory response induced by alpha-bungarotoxin.  

PubMed

Postcapillary venules represent the segment of the microvasculature most vulnerable to inflammatory processes. While there is a considerable body of data on the peripheral vasculature, little is known about the primary events occurring during inflammatory reactions in cerebral blood vessels. We introduce here a model by which the migration of polymorphonuclear leukocytes through the CNS endothelial barrier can be studied. Alpha-bungarotoxin is used as a chemotactic agent and is shown, for the first time, to act by activating the complement cascade. Leukocytes migrate through the endothelium transcellularly. Two modes of migration are described: a direct mode whereby the cells use temporary pores in the vessel wall as portals, and an indirect mode whereby the leukocytes leave the vascular compartment after being enveloped by and incorporated into endothelial cells. The functional implications of these findings lead us to conclude that the direct mode of migration is a causal agent in the massive breakdown of the blood-brain barrier under acute inflammatory conditions. PMID:4053172

Faustmann, P M; Dermietzel, R

1985-01-01

18

Chemoattractant Receptor Functions in Human Polymorphonuclear Leukocytes are Divergently Altered by Membrane Fluidizers  

Microsoft Academic Search

The chemotactic factor receptor on leukocytes initiates several cellular responses including chemotaxis, lysosomal enzyme secretion, and O2- production. The latter two responses require approximately 10-100 times more chemoattractant than is required for chemotaxis. We determined the effects of membrane fluidizers on the binding characteristics and the functional activities of the oligopeptide fMet-Leu-Phe chemotactic factor receptor on polymorphonuclear leukocytes. Fluidization was

Itzhak Yuli; Akimitsu Tomonaga; Ralph Snyderman

1982-01-01

19

Isolation and Properties of Phagocytic Vesicles from Polymorphonuclear Leukocytes  

PubMed Central

A method for the isolation of intact phagocytic vesicles from guinea pig peritoneal-exudate granulocytes and human peripheral-blood leukocytes is presented. After leukocytes ingested the particles of a stable emulsion of paraffin oil, the uningested emulsion was washed away and the cells were homogenized. The homogenate was placed in the middle of a three-step discontinuous sucrose gradient and centrifuged for 1 hr at 100,000 g. The phagocytic vesicles, containing the low density paraffin-oil particles, were simultaneously washed and collected by floatation, while the other organelles, chiefly granules, sedimented through the lower wash layer, and the particle-free supernatant remained in the middle of the gradient. Emulsion particles stained with Oil Red O were employed to assay the rate of phagocytosis and to mark the location of the particles in subcellular fractions. The dye was extracted from washed cells or cell fractions with dioxane and colorimetrically quantified. The purity of phagocytic vesicles obtained by this method was assessed by electron microscopy, chemical analysis, and assay of enzyme composition. Granule-associated enzymes, acid phosphatase, alkaline phosphatase, ?-glucuronidase, and peroxidase were present in the phagocytic vesicles and originated from the granules. Cyanide-resistant NADH (reduced form of diphosphopyridine nucleotide) oxidase was also found. Enzymes associated with the vesicles exhibited latency to Triton X-100. Uptake of particles and the transfer of total protein and phospholipid into phagocytic vesicles occurred simultaneously Accumulation of acid and alkaline phosphatase in the vesicles continued until phagocytosis ceased. Peroxidase, NADH oxidase, and ?-glucuronidase activities in the phagocytic vesicles, on the other hand, were maximal by 30 min and increased little thereafter even when phagocytosis was still going on. Images PMID:4106463

Stossel, Thomas P.; Pollard, Thomas D.; Mason, Robert J.; Vaughan, Martha

1971-01-01

20

Direct and indirect effects of E. Coli lipopolysaccharide on isolated human polymorphonuclear granulocytes and mixed leukocytes  

Microsoft Academic Search

Polymorphonuclear neutrophil granulocytes (PMN) may contribute to the lung injury induced by nonpulmonary infections with gram-negative bacteria. The direct effect ofE. coli lipopolysaccharide (LPS) on isolated human PMN or mixed leukocytes (ML), as well as the priming effect of preincubating cells with LPS, was examined in assays measuring the maximal rate of oxygen consumption (OC), cell chemiluminescence (CHML), and aggregation

Helge Opdahl

1993-01-01

21

Cigarette Smoking Causes Sequestration of Polymorphonuclear Leukocytes Released from the Bone Marrow in Lung Microvessels  

Microsoft Academic Search

Studies from our laboratory have shown that chronic cigarette smoke exposure causes a neutrophilia asso- ciated with a shortening of the mean transit time of polymorphonuclear leukocytes (PMN) though the post- mitotic pool of the marrow. The present study was designed to test the hypothesis that PMN newly re- leased from bone marrow by smoke exposure preferentially sequestered in pulmonary

Takeshi Terashima; Maria E. Klut; Dean English; Jennifer Hards; James C. Hogg; Stephan F. van Eeden

22

Experimental bacterial pneumonia in rabbits: polymorphonuclear leukocyte margination and sequestration in rabbit lungs and quantitation and kinetics of /sup 51/Cr-labeled polymorphonuclear leukocytes in E. coli-induced lung lesions  

SciTech Connect

A relationship between the circulating and marginal polymorphonuclear leukocyte (PMN) pools was documented using /sup 51/Cr-labeled leukocytes as a marker. /sup 51/Cr-leukocytes marginating in the lungs were found to decrease following a first-order exponential decline, while /sup 51/Cr radioactivity accumulated in the liver and the spleen. Intravenously administered endotoxin caused a rapid selective disappearance of PMNs from the circulation. The percentage of infused /sup 51/Cr cells disappearing was equal to the percentage of disappearance of host cells. The PMNs were found to sequester in the lungs, with peak sequestration of labeled cells occurring 5 min after an endotoxin challenge. Over the next 25 min the /sup 51/Cr radioactivity in the lungs declined. Large numbers of PMNs, probably newly derived from the bone marrow, were observed histologically to be sequestered in the lung vasculature 90 min after an endotoxin dose, while the early sequestration of circulating leukocytes could not be assessed histologically. Pulmonary inflammatory lesions were induced selectively with Escherichia coli in the left lower lobes of rabbits, leaving the right lower lobes as intrinsic controls. PMN-accumulation into the lesions was quantitated using /sup 51/Cr-labeled blood leukocytes. With the aid of /sup 125/I-labeled E. coli, a logarithmic dose-response relationship was found between the number of E. coli and of PMNs. Over a 6-hr period circulating PMNs were found to accumulate in a lesion in the left lower lobe, whereas in the control right lower lobe, leukocyte radioactivity declined. These findings were confirmed with the aid of lavages of the right and left lungs. Two peaks of PMN-accumulation were found by studying leukocyte kinetics: a larger peak between 0 and 6 hr and a smaller peak 18-24 hr after instillation of the microorganisms. Histologic studies confirmed the accumulation of leukocytes, and by 3 weeks showed a complete resolution of the lesions.

Cybulsky, M.I.; Movat, H.Z.

1982-12-01

23

Impaired metabolic function of polymorphonuclear leukocytes in glycogen storage disease Ib  

Microsoft Academic Search

To elucidate the basis for the recurrent infections in patients with glycogen storage disease (GSD) Ib we tested polymorphonuclear leukocyte (PMN) function in one patient. Bactericidal capacity and phagocytosis-induced O2 consumption were reduced. Also, phorbol myristate acetate-stimulated superoxide production and glucose oxidation through the hexose monophosphate shunt were diminished compared to control subjects. Therefore it could be speculated that in

M. Gahr; K. Heyne

1983-01-01

24

Crocidolite-induced reactive oxygen metabolites generation from human polymorphonuclear leukocytes.  

PubMed

In order to study the mechanism of carcinogenicity of crocidolite asbestos, we have investigated the species of reactive oxygen metabolites (ROM) induced by crocidolite from human polymorphonuclear leukocytes (PMN) utilizing both an electron spin resonance (ESR) spin trapping method with 5,5-dimethyl-1-pyrroline N-oxide (DMPO), and a luminol-dependent chemiluminescence (CL) method. The present study confirms the generation of OH. from human peripheral blood PMN stimulated by UICC crocidolite utilizing ESR. In addition, PMN incubated with 25-400 micrograms/ml of crocidolite produced CL, the intensity of CL increasing in a dose-dependent manner. Superoxide dismutase, catalase, and dimethyl sulfoxide, which are scavengers of O2-, H2O2, and OH., respectively, inhibited the production of crocidolite-stimulated CL from PMN, also in a dose-dependent manner. Sodium azide, an inhibitor of myeloperoxidase (MPO) to produce OCl-, also inhibited CL production. These results suggest the involvement of O2-, H2O2, OH., and OCl- in the production of CL by crocidolite-stimulated PMN. In conclusion, it is proposed that OH. is a key ROM species in the mechanism of crocidolite-induced carcinogenesis. PMID:8055842

Ishizaki, T; Yano, E; Urano, N; Evans, P H

1994-08-01

25

Relationship of polymorphonuclear leukocytes to capillary dropout in the human diabetic choroid.  

PubMed Central

Capillary dropout is an initial event in diabetic retinopathy, but the etiology is unknown. Recent evidence suggests that similar events may occur in the diabetic choroid. We have developed a method to evaluate the relationship between the polymorphonuclear leukocytes (PMNs) and capillary dropout in the human diabetic choroid using alkaline phosphatase (APase) histochemistry to label blood vessels and nonspecific esterase activity to identify PMNs. The number and distribution of PMNs in diabetic and nondiabetic choroidal capillaries (choriocapillaris) were analyzed in the flat perspective and the tissue then flat embedded in glycol methacrylate for histological sectioning. The total number of PMNs was increased within the choriocapillaris in five diabetic eyes (170.9 +/- 12.9 PMNs/mm2 of choroid) compared with five nondiabetic eyes (84.2 +/- 16.9 PMNs/mm2; P < 0.001). PMNs were almost always within blood vessel lumens and not in interstitial tissue. In the diabetic choroid, increased numbers of PMNs were present in areas of choriocapillaris with pathological changes (loss in APase activity and choroidal neovascularization) compared with nonpathological choriocapillaris (205.1 +/- 46.9 PMNs/mm2 in pathological versus 152.3 +/- 23.4 PMNs/mm2 in nonpathological areas; P < 0.001). PMNs were often queued up within the lumens of capillaries, demonstrating loss in APase activity. We have observed an increased number of PMNs in diabetic choroid compared with control nondiabetic choroids, and PMNs in diabetic choroid were associated with loss in APase activity, which was related to loss in viable endothelial cells. The results suggest that PMNs contribute to vaso-occlusive processes and endothelial cell injury in the diabetic choroid. Images Figure 1 Figure 2 Figure 3 PMID:9284819

Lutty, G. A.; Cao, J.; McLeod, D. S.

1997-01-01

26

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

PubMed Central

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

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

2011-01-01

27

Leukotriene D4 activates {beta}2-integrin adhesion in human polymorphonuclear leukocytes.  

PubMed

We examined the functional role and mechanisms by which activation of cysteinyl leukotriene-1 receptor (cysLT(1)R) regulates beta(2)-integrin adhesion to intercellular adhesion molecule (ICAM)-1 in human polymorphonuclear leukocytes (PMNs) in vitro. Human peripheral blood PMNs and eosinophils were isolated separately from the same mildly atopic donors. Surface expression of cysLT(1)R was identified both in PMNs and in eosinophils by immunofluorescence analysis. Total cysLT(1)R protein was substantially greater in eosinophils than in PMNs as determined by Western blot analysis. However, leukotriene D(4) (LTD(4)) upregulated beta(2)-integrin adhesion of PMNs to ICAM-1 with high efficacy in a time- and concentration-dependent manner. Upregulated beta(2)-integrin adhesion of PMNs was related temporally and quantitatively to phosphorylation of 85-kDa cytosolic group IVa phospholipase A2 (gIVaPLA2). Augmented LTD(4)-induced adhesion was blocked significantly by montelukast, a cysLT(1)R antagonist. Trifluoromethylketone (a gIVaPLA2 inhibitor) blocked beta(2)-integrin adhesion caused by LTD(4) activation, as did anti-CD18 monoclonal antibody directed against beta(2)-integrin on the PMN surface. Our data demonstrate that LTD(4) causes phosphorylation of gIVaPLA2 and upregulation of beta(2)-integrin adhesion to ICAM-1 or ICAM-1 surrogate through cysLT(1)R activation. Activation of gIVaPLA2 is a critical step through which beta(2)-integrin adhesion is upregulated by the cysLT(1)R expressed on the surface membrane of human PMN. PMID:19679609

Meliton, A Y; Muñoz, N M; Osan, C M; Meliton, L N; Leff, A R

2010-02-01

28

Impaired metabolic function of polymorphonuclear leukocytes in glycogen storage disease Ib.  

PubMed

To elucidate the basis for the recurrent infections in patients with glycogen storage disease (GSD) Ib we tested polymorphonuclear leukocyte (PMN) function in one patient. Bactericidal capacity and phagocytosis-induced O2 consumption were reduced. Also, phorbol myristate acetate-stimulated superoxide production and glucose oxidation through the hexose monophosphate shunt were diminished compared to control subjects. Therefore it could be speculated that in PMN of patients with GSD Ib, glucose-6-phosphate has no access to the enzymes of the hexose monophosphate shunt due to a transport-related defect as shown for glucogenesis in hepatocytes. PMID:6578930

Gahr, M; Heyne, K

1983-09-01

29

Polymorphonuclear leukocytes from patients with severe sepsis have lost the ability to degrade fibrin via u-PA.  

PubMed

Fibrin persistence in the vasculature is an important complication of sepsis that can often lead to mortality. We have previously established that polymorphonuclear leukocytes (PMN) from healthy individuals have the capacity to degrade fibrin via urokinase-type plasminogen activator (u-PA). We have also demonstrated an increase in u-PA antigen in the plasma of patients suffering from septic shock. In this study, we investigate the hypothesis that PMN from patients with sepsis have lost their fibrinolytic ability and that this might contribute to the persistence of fibrin deposits. We show here that PMN from these patients do not express any u-PA activity, despite retaining some u-PA antigen. Additionally, thrombi prepared from the whole blood of the patients exhibit reduced endogenous lysis compared with those from healthy individuals. These data indicate that loss of fibrinolytic activity from PMN may be a contributing factor in fibrin persistence in the microvasculature in sepsis. PMID:15277568

Moir, E; Greaves, M; Adey, G D; Bennett, B

2004-09-01

30

Endothelin1 Changes Polymorphonuclear Leukocytes' Deformability and CD11b Expression and Promotes Their Retention in the Lung  

Microsoft Academic Search

Endothelin (ET)1 influences polymorphonuclear leukocyte (PMN)- endothelial cell interactions. The aim of this study was to ex- amine the effect of ET-1 on factors that influence PMN-endo- thelial interaction and retention in the lung both in vitro and in vivo. In vitro , high concentration of ET-1 ( > 10 2 8 M) rapidly increased PMN F-actin content (10 2

Yukio Sato; James C. Hogg; Dean English; Stephan F. van Eeden

31

Human polymorphonuclear leukocytes are attracted to the anode in a weak electric field  

SciTech Connect

A modified Boyden chamber was used to measure the galvanotaxis of human polymorphonuclear leukocytes (PMN) in a constant dc electric field. In these experiments the PMN were placed onto the top surface of a cellulose nitrate filter (5um) and the electrode of interest was placed under the lower surface. The numbers of PMN's on lower surface were counted with controls. More PMN migrated to the anode and less to the cathode when compared to controls. Because thermal heating is identical when either electrode is used, heating is not thought to be a significant factor in increasing the non-directional migration of PMN. When the temperature at which the experiments are conducted was lowered from 37C to 25C or lower, there was no increase in the number of PMN on the anode surface. This indicates the migration of PMN to the anode is not due to a passive electrophoretic phenomena but due to a metabolically dependent activity of the PMN.

Hamada, S.; Davis, V. III; Robinson, A.; Allen, K. (West Georgia College, Carrollton (United States))

1991-03-15

32

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

SciTech Connect

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.

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

1984-04-01

33

Degradation of Chlamydia trachomatis in human polymorphonuclear leukocytes: an ultrastructural study of peroxidase-positive phagolysosomes.  

PubMed Central

We have previously shown that human polymorphonuclear leukocytes (PMNs) killed organisms belonging to both human biovars of Chlamydia trachomatis. However, the mechanism of destruction was still unclear. We therefore conducted an ultrastructural and cytochemical study to investigate the mechanism of chlamydial degradation. PMNs were inoculated with the trachoma serovar B (B/TW-5/OT) or with the lymphogranuloma venereum serovar L2 (L2/434/Bu) for 15, 30, 60, or 120 min and then fixed and processed for transmission electron microscopy. Diaminobenzidine, a cytochemical marker, was used to demonstrate the localization of intracellular peroxidase. Ultrastructural evidence is presented showing the progressive degradation of chlamydiae over a 2-h period within peroxidase-positive phagolysosomes. Pretreatment of organisms with normal or immune serum was not required for the process of degradation. Images PMID:3015802

Yong, E C; Chi, E Y; Chen, W J; Kuo, C C

1986-01-01

34

Differential inhibition of polymorphonuclear leukocyte recruitment in vivo by dextran sulphate and fucoidan  

PubMed Central

The selectin-mediated rolling of leukocytes along the endothelial cells is a prerequisite step followed by firm adhesion and extravasation into the inflamed tissue. This initial contact can be suppressed by sulphated polysaccharides. We have studied the effect of sulphated polysaccharides on the ultimate polymorphonuclear leukocyte (PMN) recruitment and plasma leakage in rabbit skin in response to intradermal injection of various inflammatory mediators. PMN infiltration evoked by various PMN chemoattractants (FMLP, C5a desArg, LTB4 and IL-8) was significantly inhibited after intravenous injection of dextran sulphate (25 mg/kg), heparin (2 × 90 mg/kg) or fucoidan (1 mg/kg). PMN-dependent plasma leakage was equally well reduced by the different sulphated polymers. Vascular permeability induced by histamine or thrombin acting via a PMN-independent mechanism was not reduced. Fucoidan was the only polysaccharide able to suppress IL-1-induced PMN infiltration for 60–70%. Local administration of dextran sulphate had no effect on PMN-dependent plasma leakage. Differential inhibition of PMN recruitment was determined after injection of dextran sulphate or fucoidan depending on the type of insult. Therefore, these results suggest that different adhesion pathways are utilized during PMN recruitment in vivo in response to chemoattractants and IL-1. PMID:18475729

Rampart, M.; Herman, A. G.

1996-01-01

35

Polymorphonuclear Leukocyte Bactericidal Activity and Oxidative Metabolism During Glutathione Peroxidase Deficiency  

PubMed Central

Glutathione peroxidase (GPx) deficiency has been proposed as a cause of some instances of chronic granulomatous disease (CGD). GPx activity varies greatly among species, and specific deficiency of this selenium-dependent enzyme can be produced by dietary selenium deficiency in rats. Bactericidal activity of polymorphonuclear (PMN) leukocytes from normal rats, humans, and guinea pigs (GPx high, intermediate, and nearly absent, respectively), selenium-deficient rats (GPx absent), and a patient with CGD were compared. There was no correlation between natural levels of GPx and bactericidal activity; only CGD was associated with inability to kill a Proteus mirabilis strain in vitro (killing known to be dependent on oxidative mechanisms). Postphagocytic metabolism was examined in normal and GPx-deficient rats. Both demonstrated normal iodination and superoxide production during phagocytosis and gave similar histochemical reduction of nitroblue tetrazolium dye under either resting or endotoxin-stimulation conditions. Postphagocytic hexose monophosphate shunt activity was somewhat lower in PMN from GPx-deficient animals as compared with normal but was substantially (10-fold) higher than that observed in resting cells. Thus, postphagocytic oxidative responses and subsequent bactericidal activity of PMN leukocytes were not compromised by complete absence of GPx, even in the species with the highest natural level of this enzyme. These results are not compatible with the hypothesis that CGD can be caused by a deficiency of GPx. PMID:198376

Bass, D. A.; DeChatelet, L. R.; Burk, R. F.; Shirley, P.; Szejda, P.

1977-01-01

36

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

PubMed

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

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

2000-01-01

37

Manno(Rhamno)Biose-Containing Capsular Polysaccharides of Klebsiella pneumoniae Enhance Opsono-Stimulation of Human Polymorphonuclear Leukocytes  

Microsoft Academic Search

We tested the relationship between the capsular and the O-antigen structures and the ability of bacteria to trigger respiratory burst in human polymorphonuclear leukocytes (PMNL).Capsulated and non-capsulated variants as well as capsule-switched derivatives of Klebsiella serotypes bearing or lacking manno(rhamno)biose repeats in their capsular polysaccharides and expressing either mannose-rich or mannose-poor O antigens were tested for their ability to induce

Hany Sahly; Yona Keisari; Itzhak Ofek

2009-01-01

38

Effects of serine protease inhibitors on accumulation of polymorphonuclear leukocytes in the lung induced by acute pancreatitis in rats  

Microsoft Academic Search

The administration of a high-dose of a serine protease inhibitor is recommended in patients complicated by multiple organ\\u000a failure (MOF), including adult respiratory distress syndrome (ARDS), induced by acute pancreatitis. The accumulation of polymorphonuclear\\u000a leukocytes (PMN) in affected organs is considered to be one of the causative factors of MOF. Adhesion to endothelial cells\\u000a (EC), via adhesion molecules, and the

Yoshiaki Okumura; Hisayuki Inoue; Yoshihide Fujiyama; Tadao Bamba

1995-01-01

39

Polymorphonuclear leukocyte adherence induces actin polymerization by a transduction pathway which differs from that used by chemoattractants  

Microsoft Academic Search

Abstract. Nitrobenzoxadiazole-phallacidin in combina- tion with quantitative fluorescent microscopy,have been used to measure,F-actin concentrations,in human polymorphonuclear leukocytes (PMN) as they adhere to a plastic surface. Like stimulation with chemoat- tractants, adherence is associated with a twofold rise in F-actin content. However unlike the rapid rise in F-actin induced by chemoattractants which,peaks within 30 s, actin assembly induced by adherence is

Frederick S. Southwick; Guissou A. Dabiri; Miriam Paschetto; Sally H. Zigmondt

1989-01-01

40

Complement and polymorphonuclear leukocytes do not determine the vascular permeability induced by intraocular LPS.  

PubMed

The intravitreous injection of an endotoxin of Escherichia coli 055:B5 (LPS; 0.1-0.5 microgram/50 microliters of saline) induces ocular inflammation in rabbits that is maximal 20-24 hours later and disappears by 4 days. The inflammation is characterized by an alteration in ocular vascular permeability (OVP) measured by the ocular extravasation of 125I-albumin and an outpouring of leukocytes, most of which are polymorphonuclear leukocytes (PMNs), as determined by histopathologic study. Nitrogen mustard (mechlorethamine, 1.75 mg/kg) administered 3 days prior to LPS virtually eliminates PMNs in the circulation and those infiltrating ocular tissues 20 hours after intravitreous LPS, and yet the average increase in vascular permeability is not different from that of controls. Cobra venom factor (CVF; 300-400 units) 7 hours before intravitreous LPS produces a greater than 90% decrease in both hemolytic complement activity and zymosan-inducible serum chemotactic activity; yet 20 hours after LPS, the OVP is the same in CVF-treated rabbits and controls. For comparison, an ocular passive Arthus reaction (ovalbumin-anti-ovalbumin) was significantly affected by CVF pretreatment. Chemotactic activity in the aqueous humor is found in both CVF-treated and control rabbits 20 hours after intravitreous LPS. This activity attracts rabbit, but not human, PMNs, is partially heat-sensitive, and is not inhibited when PMNs are preincubated with C5a. These results indicate that neither PMNs nor circulating complement determine the OVP following intravitreous LPS, and that the chemotactic activity present in aqueous humor at the height of the inflammatory response is not primarily C5a. PMID:3155595

Howes, E L; Wong, K L; Hartiala, K T; Webster, R O; Rosenbaum, J T

1985-01-01

41

Complement and polymorphonuclear leukocytes do not determine the vascular permeability induced by intraocular LPS.  

PubMed Central

The intravitreous injection of an endotoxin of Escherichia coli 055:B5 (LPS; 0.1-0.5 microgram/50 microliters of saline) induces ocular inflammation in rabbits that is maximal 20-24 hours later and disappears by 4 days. The inflammation is characterized by an alteration in ocular vascular permeability (OVP) measured by the ocular extravasation of 125I-albumin and an outpouring of leukocytes, most of which are polymorphonuclear leukocytes (PMNs), as determined by histopathologic study. Nitrogen mustard (mechlorethamine, 1.75 mg/kg) administered 3 days prior to LPS virtually eliminates PMNs in the circulation and those infiltrating ocular tissues 20 hours after intravitreous LPS, and yet the average increase in vascular permeability is not different from that of controls. Cobra venom factor (CVF; 300-400 units) 7 hours before intravitreous LPS produces a greater than 90% decrease in both hemolytic complement activity and zymosan-inducible serum chemotactic activity; yet 20 hours after LPS, the OVP is the same in CVF-treated rabbits and controls. For comparison, an ocular passive Arthus reaction (ovalbumin-anti-ovalbumin) was significantly affected by CVF pretreatment. Chemotactic activity in the aqueous humor is found in both CVF-treated and control rabbits 20 hours after intravitreous LPS. This activity attracts rabbit, but not human, PMNs, is partially heat-sensitive, and is not inhibited when PMNs are preincubated with C5a. These results indicate that neither PMNs nor circulating complement determine the OVP following intravitreous LPS, and that the chemotactic activity present in aqueous humor at the height of the inflammatory response is not primarily C5a. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 PMID:3155595

Howes, E. L.; Wong, K. L.; Hartiala, K. T.; Webster, R. O.; Rosenbaum, J. T.

1985-01-01

42

Filter Buffy Coats (FBC): A source of peripheral blood leukocytes recovered from leukocyte depletion filters  

Microsoft Academic Search

In compliance with federal regulations, blood banks routinely use leukocyte depletion filters to eliminate contaminating leukocytes from blood products such as red blood cell and platelet concentrates. We developed and optimized conditions to elute leukocytes adsorbed to these filters; resulting in leukocyte suspensions which we termed Filter Buffy Coats (FBCs). These Filter Buffy Coats can replace standard buffy coats for

T. P. H. Meyer; I. Zehnter; B. Hofmann; J. Zaisserer; J. Burkhart; S. Rapp; F. Weinauer; J. Schmitz; W. E. Illert

2005-01-01

43

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

PubMed Central

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

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

1987-01-01

44

Burkholderia pseudomallei activates complement and is ingested but not killed by polymorphonuclear leukocytes.  

PubMed Central

The mechanism by which Burkholderia pseudomallei is resistant to lysis by human serum is unknown but may include interference with complement activation, effective opsonization, or complement-mediated lysis. We investigated the interaction of B. pseudomallei with complement in the presence and absence of specific antibody to determine potential mechanisms of serum resistance. We demonstrated rapid activation and consumption of complement by B. pseudomallei which, in the absence of specific antibody, occurred predominantly via the alternative pathway. Complement activation was associated with deposition of the opsonically active C3b and iC3b fragments on the bacterial surface. C5b-9, detected on the bacterial surface after opsonic periods of 1 to 60 min, was susceptible to elution by 1 M NaCl, indicating that resistance to complement-mediated lysis may result from deposition of the membrane attack complex in a nonmicrobicidal location. To define the role of opsonins, we investigated the ability of polymorphonuclear leukocytes (PMNL) to phagocytose B. pseudomallei. Phagocytosis of bacteria by PMNL, and the observed oxidative response, was significantly increased by opsonization of organisms with complement and/or specific antibody. Despite opsonophagocytosis by PMNL and the production of an oxidative response, no significant bacterial killing was observed. PMID:8945532

Egan, A M; Gordon, D L

1996-01-01

45

Intracellular penetration and activity of BAY Y 3118 in human polymorphonuclear leukocytes.  

PubMed Central

The penetration of a new quinolone (BAY Y 3118) into human polymorphonuclear leukocytes (PMNs) was evaluated by a fluorometric assay. The cellular concentration-to-extracellular concentration (C/E) ratio was higher than 6.3 at extracellular concentrations ranging from 2 to 100 mg/liter. The uptake of BAY Y 3118 was rapid, reversible and nonsaturable. The intracellular penetration of BAY Y 3118 was significantly affected by environmental temperature (C/E ratio at 4 degrees C, 5.4 +/- 0.5; control, 7.5 +/- 0.9; P < 0.05) and cell viability (C/E ratio in dead PMNs, 5.5 +/- 0.8; control 7.5 +/- 0.9; P < 0.05), but it was not affected by metabolic inhibitors. The ingestion of opsonized zymosan or opsonized Staphylococcus aureus significantly decreased the levels of PMN-associated BAY Y 3118. Cell stimulation by a membrane activator, however, significantly increased the intracellular concentration of this quinolone. At therapeutic extracellular concentrations (0.5, 2, and 5 mg/liter), BAY Y 3118 showed intracellular activity greater than that of ciprofloxacin against S. aureus in human PMNs. It was concluded that BAY Y 3118 reaches high intracellular concentrations within human PMNs and remains active intracellularly. PMID:7840582

Garcia, I; Pascual, A; Perea, E J

1994-01-01

46

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

PubMed

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

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

2014-08-01

47

Specific binding of leukotriene B4 to a receptor on human polymorphonuclear leukocytes  

PubMed Central

In this paper we have described the binding of nanomoler concentrations of [3H]leukotriene B4 (LTB4) to human polymorphonuclear leukocytes. Because up to 80% of the total [3H]LTB4 binding was blocked by excess (greater than 100 times) [14C]LTB4, the majority of binding is specific. Stereospecificity of the LTB4 binding is demonstrated by the diminished relative abilities of the 6-trans-and 12-epi-6-trans- isomers of LTB4 to block [3H]LTB4 binding. With these two isomers 3-10- fold higher than [14C]LTB4 concentrations were needed for equivalent inhibition of [3H]LTB4 binding. This difference is quantitatively less dramatic than the differences between these isomers in many in vitro functional assays such as chemokinesis, chemotaxis, and degranulation. Binding of [3H]FMLP is not blocked at greater than 100-fold excess of LTB4. The binding of [3H]LTB4 to cells appears to be essentially irreversible at 4 degrees C, but not at 37 degrees C where initially bound LTB4 is rapidly converted to metabolites which then enter the medium. These results suggest the presence of a saturable, stereospecific site for LTB4 on PMN. The association of LTB4 binding and the initiation of pharmacological responses to LTB4 will require further studies. PMID:6296265

1983-01-01

48

Induction of 5-lipoxygenase activation in polymorphonuclear leukocytes by 1-oleoyl-2-acetylglycerol.  

PubMed

1,2-Diacylglycerols (DAGs) can prime polymorphonuclear leukocytes (PMNL) for enhanced release of arachidonic acid (AA) and generation of 5-lipoxygenase (5-LO) products upon subsequent agonist stimulation. Here, we demonstrate that in isolated human PMNL, 1-oleoyl-2-acetylglycerol (OAG) functions as a direct agonist stimulating 5-LO product formation (up to 42-fold). OAG caused no release of endogenous AA, but in the presence of exogenous AA, the magnitude of 5-LO product synthesis induced by OAG was comparable to that obtained with the Ca(2+)-ionophore A23187. Interestingly, OAG-induced 5-LO product synthesis was not connected with increased 5-LO nuclear membrane association. Examination of diverse glycerides revealed that the sn-2-acetyl-group is important, thus, also 1-O-hexadecyl-2-acetylglycerol (EAG) stimulated 5-LO product formation (up to 8-fold). Treatment of PMNL with OAG did not alter the mobilization of Ca(2+) but removal of intracellular Ca(2+) abolished the upregulatory OAG effects. Notably, the PKC activator phorbol-myristate-acetate hardly increased 5-LO product synthesis and PKC inhibitors failed to suppress the effects of OAG. Although OAG rapidly activated p38 MAPK and p42/44(MAPK), which can stimulate 5-LO for product synthesis, specific inhibitors of these kinases could not prevent 5-LO activation by OAG. Together, OAG acts as a direct agonist for 5-LO product synthesis in PMNL stimulating 5-LO by novel undefined mechanisms. PMID:12573453

Albert, Dana; Buerkert, Eva; Steinhilber, Dieter; Werz, Oliver

2003-02-20

49

Localization of electron-dense tracers during entry of Rickettsia tsutsugamushi into polymorphonuclear leukocytes.  

PubMed

The invasion of Rickettsia tsutsugamushi, Gilliam strain, into guinea pig polymorphonuclear leukocytes (PMNs) and the localization and distribution of tracers were followed during the process by electron microscopy. The seven tracers used were: cationized ferritin, ferritin, thorium dioxide (ThO2), carbon particles, latex spheres, paraffin oil, and Escherichia coli. These markers were added to the incubation medium containing the PMNs before or simultaneously with R. tsutsugamushi-infected BHK-21 cells. Both morphologically intact and degenerating rickettsiae were present in the phagosomes in PMNs, but only the viable-appearing rickettsiae were free in the cytoplasm. The intact rickettsiae were singly and selectively phagocytized in tightly enclosed phagosomal membranes which usually excluded the tracers, except when ThO2 or ferritin was used. When ThO2, which labels the plasma membrane of PMNs, was used. ThO2-labeled phagosomal membranes enclosing rickettsiae were observed and short membrane fragments still labeled with this tracer were found in the vicinity of rickettsiae in the cytoplasmic matrix of PMNs. When ferritin or ThO2 was used as a tracer, some of the phagosomes contained rickettsiae still enclosed in an envelope of BHK-21 cytoplasm and cell membrane. Phagolysosomes preloaded with electron-dense markers fused with subsequently formed phagosomes containing degenerated rickettsiae but not with those containing intact rickettsiae. These results support our interpretation that viable rickettsial entry into PMNs is by selective phagocytosis and escape from these phagosomes. PMID:6777300

Rikihisa, Y; Ito, S

1980-10-01

50

Membrane-bound lactoferrin alters the surface properties of polymorphonuclear leukocytes.  

PubMed Central

Polymorphonuclear leukocytes (PMN) aggregate and avidly attach to endothelium in response to chemotactic agents. This response may be related in part to the release of the specific granule constituent lactoferrin (LF). We found by using immunohistology and biochemical and biophysical techniques that LF binds to the membrane and alters the surface properties of the PMN. Upon exposure of PMN treated with 5 micrograms/ml cytochalasin B to 2 x 10(-7) M formyl-methionine-leucine-phenylalanine for 5 min, the PMN mobilized LF to their surface as observed by immunoperoxidase staining for LF. At added LF levels ranging from 4 to 15 micrograms/10(7) PMN there was a dose-dependent reduction in PMN surface charge reaching 4 mV, when the partitioning into the membrane of a charged amphipathic nitroxide spin label was measured by electron spin resonance spectroscopy, whereas transferrin was without effect. When 125I-FeLF was added to human PMN in increasing amounts and the results corrected for the residual amount of free LF contaminating the cells, the PMN were saturated with LF at concentrations between 100 and 200 nM in the medium. Human PMN bound 1.35 x 10(6) molecules per cell and the calculated value for the association constant for these receptors was 5.2 x 10(6) M-1. Additionally, 6 micrograms/ml LF served as an opsonin for rabbit MN to promote PMN uptake by rabbit macrophages, when assessed by electron microscopy, but lysozyme did not. These studies indicate that LF can bind to the surface of the PMN and reduce its surface charge. This correlates with enhanced "stickiness" leading to a variety of cell-cell interactions. Images PMID:6290534

Boxer, L A; Haak, R A; Yang, H H; Wolach, J B; Whitcomb, J A; Butterick, C J; Baehner, R L

1982-01-01

51

Stimulus-specific effects of endotoxin on superoxide production by rabbit polymorphonuclear leukocytes.  

PubMed Central

The release of superoxide (O2-) by polymorphonuclear leukocytes (PMN) is an important function that contributes to microbial death. Controversy exists as to the effect of bacterial endotoxin (lipopolysaccharide, or LPS) on the production of O2-. We have injected rabbits with 25 micrograms Escherichia coli LPS intravenously and studied PMN function 18 to 24 hours later. Relative to PMN from saline-injected controls, PMN from LPS-treated rabbits released markedly greater amounts of O2- in response to 10 ng/ml phorbol myristate acetate (PMA) as measured by nmol cytochrome C reduced in 20 minutes (40.8 +/- 7.8 for LPS-treated PMN versus 10.1 +/- 1.6 for control, p less than 0.01). LPS injection, however, significantly reduced O2- release in response to C (complement) 5a (1.4 +/- 0.6 nmole/20 minutes for LPS-treated PMN versus 5.6 +/- 1.3 nmole/20 minutes for control, p less than 0.01). O2- release in response to a third stimulus, n-formyl-methionyl-leucyl-phenylalanine (10(-7) to 10(-9) M), was not affected by LPS. O2- release in response to PMA was enhanced over a wide range of PMA concentrations (10 to 300 ng/ml). Kinetic studies over 30 minutes indicated that, after a brief initial latency in measurable response, LPS enhanced responsiveness to PMA at all time points observed. The reduced responsiveness to C5a corresponds to a previously reported down regulation of receptors for this ligand after intravenous LPS. The observations indicate that intravenous LPS can alter a critical function of PMN for at least 24 hours in a stimulus-specific manner. PMID:2827396

Rosenbaum, J. T.; Enkel, H.

1987-01-01

52

Effects of hypertonic saline on expression of human polymorphonuclear leukocyte adhesion molecules.  

PubMed

Hypertonic saline prevents vascular adherence of neutrophils and ameliorates ischemic tissue injury. We hypothesized that hypertonic saline attenuates N-formyl-methionyl-leucyl-phenylalanine (fMLP)-stimulated expression of adhesion molecules on human polymorphonuclear leukocytes (PMNLs). fMLP-stimulated up-regulation of beta2-integrins was diminished by hypertonic saline but not by hypertonic choline chloride-, mannitol-, or sucrose-modified Hanks' buffered salt solution. Shedding of L-selectin was decreased by hypertonic saline and choline chloride but not by hypertonic mannitol or sucrose. When the effects of hypertonic sodium chloride- and choline chloride-modified media were compared, neither solution affected fMLP-receptor binding but both equally inhibited fMLP-stimulated increase in intracellular calcium, ionophore A23187, and phorbol myristate acetate (PMA)-stimulated numerical up-regulation of beta2-integrins. Analysis of mitogen-activated protein (MAP) kinases p38 and p44/42 for phosphorylation revealed that hypertonic solutions did not differ in preventing fMLP-stimulated increases in phospho-p38 and phospho-p44/42. Resting PMNLs shrunk by hypertonic saline increased their volume during incubation and further during chemotactic stimulation. Addition of amiloride further enhanced inhibition of up-regulation of beta2-integrins. No fMLP-stimulated volume changes occurred in PMNLs exposed to hypertonic choline chloride, resulting in significant cell shrinkage. Results suggest a sodium-specific inhibitory effect on up-regulation of beta2-integrins of fMLP-stimulated PMNLs, which is unlikely to be caused by alterations of fMLP receptor binding, decrease in cytosolic calcium, attenuation of calcium or protein kinase C-dependent pathways, suppression of p38- or p44/42 MAP kinase-dependent pathways, or cellular ability to increase or decrease volumes. PMID:11493618

Thiel, M; Buessecker, F; Eberhardt, K; Chouker, A; Setzer, F; Kreimeier, U; Arfors, K E; Peter, K; Messmer, K

2001-08-01

53

Mechanism of leukotriene generation in polymorphonuclear leukocytes by staphylococcal alpha-toxin.  

PubMed Central

The effects of staphylococcal alpha-toxin on arachidonic acid metabolism in rabbit polymorphonuclear leukocytes (PMNs) were investigated and compared with those of the ionophore A23187 and the chemotactic tripeptide formylmethionyl-leucyl-phenylalanine (fMLP). Sublytic amounts of alpha-toxin stimulated the release of leukotriene B4 (LTB4) in PMNs in a dose-dependent manner. The toxin was several times more potent than fMLP but was not as effective as the ionophore. Preincubation of the toxin with neutralizing antibodies abolished the effect. Extracellular calcium was strictly required for eliciting LTB4 generation. Verapamil, a calcium channel blocker, inhibited fMLP-mediated LTB4 generation but had no effect on alpha-toxin- or A23187-exposed PMNs. Agents such as trifluoperazine and N-6(aminohexyl)-5-chloro-1-naphthalene sulfonamid that interfered with calmodulin activity, however, inhibited LTB4 generation in all cases. One minute after the addition of alpha-toxin, PMNs exhibited a severalfold enhancement in passive permeability to 45Ca2+. In addition, these cells became permeable to sucrose but not to inulin or dextran. The influx pattern was consistent with the previous observation that alpha-toxin creates discrete transmembrane channels in erythrocytes with an effective internal diameter of 2 to 3 nm. The results suggest that alpha-toxin triggers the arachidonic acid pathway in PMNs by facilitating calcium influx into the cells, possibly via transmembrane toxin pores that serve as calcium gates. Generation of arachidonic acid metabolites in PMNs by sublytic amounts of alpha-toxin may represent an important cellular reaction that generally occurs during infections with Staphylococcus aureus. PMID:3025097

Suttorp, N; Seeger, W; Zucker-Reimann, J; Roka, L; Bhakdi, S

1987-01-01

54

Localization of electron-dense tracers during entry of Rickettsia tsutsugamushi into polymorphonuclear leukocytes.  

PubMed Central

The invasion of Rickettsia tsutsugamushi, Gilliam strain, into guinea pig polymorphonuclear leukocytes (PMNs) and the localization and distribution of tracers were followed during the process by electron microscopy. The seven tracers used were: cationized ferritin, ferritin, thorium dioxide (ThO2), carbon particles, latex spheres, paraffin oil, and Escherichia coli. These markers were added to the incubation medium containing the PMNs before or simultaneously with R. tsutsugamushi-infected BHK-21 cells. Both morphologically intact and degenerating rickettsiae were present in the phagosomes in PMNs, but only the viable-appearing rickettsiae were free in the cytoplasm. The intact rickettsiae were singly and selectively phagocytized in tightly enclosed phagosomal membranes which usually excluded the tracers, except when ThO2 or ferritin was used. When ThO2, which labels the plasma membrane of PMNs, was used. ThO2-labeled phagosomal membranes enclosing rickettsiae were observed and short membrane fragments still labeled with this tracer were found in the vicinity of rickettsiae in the cytoplasmic matrix of PMNs. When ferritin or ThO2 was used as a tracer, some of the phagosomes contained rickettsiae still enclosed in an envelope of BHK-21 cytoplasm and cell membrane. Phagolysosomes preloaded with electron-dense markers fused with subsequently formed phagosomes containing degenerated rickettsiae but not with those containing intact rickettsiae. These results support our interpretation that viable rickettsial entry into PMNs is by selective phagocytosis and escape from these phagosomes. Images Fig. 6 Fig. 7 Fig. 8 Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 9 PMID:6777300

Rikihisa, Y; Ito, S

1980-01-01

55

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

SciTech Connect

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.

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

1985-09-09

56

Altered polymorphonuclear leukocyte Fc gamma R expression contributes to decreased candicidal activity during intraabdominal sepsis  

SciTech Connect

We investigated the effects of untreated intraabdominal sepsis on polymorphonuclear leukocyte (PMN) candicidal activity. Two groups of swine were studied. Group I (n=6) underwent sham laparotomy, group II (n=7) underwent cecal ligation and incision. Untreated intraabdominal sepsis resulted in a progressive decrease in PMN candicidal activity. Concomitant rosetting and phagocytosis assays demonstrated a decrease in both the attachment and phagocytosis of Candida albicans opsonized with both normal and septic swine serum by PMNs in group II. Iodine 125-labeled swine immunoglobulin G (IgG) and fluorescein isothioalanate (FITC)-labeled swine IgG were used to investigate Fc gamma receptor ligand interactions. Scatchard analyses demonstrated a progressive decline in both the binding affinity constant and number of IgG molecules bound per PMN. Stimulation of the oxidative burst markedly reduced 125I-labeled IgG binding in both group I and group II, with a greater decrement being seen in animals with intraabdominal sepsis. Further, in group II, PMN recycling of the Fc gamma receptor to the cell surface after generation of the oxidative burst was reduced by postoperative day 4. Binding of monoclonal antibodies to Fc gamma receptor II, but not Fc gamma receptor I/III markedly reduced intracellular candicidal activity. Immunofluorescence studies revealed a homogeneous pattern of FITC-IgG uptake by nearly all group I PMNs, whereas by postoperative day 8 a substantial number of PMNs from group II failed to internalize the FITC-IgG. These studies suggest that untreated intraabdominal sepsis reduces PMN candicidal activity and that this is due, in part, to altered PMN Fc gamma receptor ligand interactions.

Simms, H.H.; D'Amico, R.; Monfils, P.; Burchard, K.W. (Rhode Island Hospital, Providence (USA))

1991-03-01

57

Function of milk polymorphonuclear neutrophil leukocytes in bovine mammary glands infected with Corynebacterium bovis.  

PubMed

Corynebacterium bovis is one of the most commonly isolated bacteria from aseptically collected bovine milk samples. The objective of the current study was to characterize the bovine innate immune response by evaluating milk polymorphonuclear neutrophilic leukocytes (PMNL) in mammary glands infected with C. bovis. Twenty quarters infected with C. bovis and 28 culture-negative quarters (with milk somatic cell count <1×10(5) cells/mL) were used. The percentages of milk PMNL and the PMNL expression of L-selectin (CD62L), ?2-integrin (CD11b), and one of the endothelial-selectin ligands (CD44), as well as the levels of intracellular reactive oxygen species (ROS) and the phagocytosis of Staphylococcus aureus, were evaluated by flow cytometry. The apoptosis and necrosis rates of the PMNL were quantified using dual-color flow cytometry with fluorescein-labeled annexin and propidium iodide. The present study revealed a higher percentage of PMNL in the milk from C. bovis-infected quarters, although no significant differences were found in levels of CD44, CD62L, or CD11b expression among the PMNL. A lower percentage of apoptotic PMNL was observed in C. bovis-infected quarters, as well as higher percentages of viable PMNL and of PMNL that produced intracellular ROS. However, no alterations were observed in phagocytosis of Staph. aureus by the PMNL or in intensity of intracellular ROS production by PMNL. Thus, results from this investigation of the PMNL function support, at least in part, the fact that intramammary infections by C. bovis may offer protection against intramammary infections by other bacteria. PMID:23608489

Blagitz, M G; Souza, F N; Santos, B P; Batista, C F; Parra, A C; Azevedo, L F F; Melville, P A; Benites, N R; Della Libera, A M M P

2013-06-01

58

Polymorphonuclear Leukocytes Restrict Growth of Pseudomonas aeruginosa in the Lungs of Cystic Fibrosis Patients.  

PubMed

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

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

2014-11-01

59

Human polymorphonuclear leukocytes produce cytokines in response to Leishmania major promastigotes.  

PubMed

Polymorphonuclear leukocytes (PMN) release cytokines that may influence the development of the subsequent adaptive immune response. Little is known about cytokines produced by human PMN in response to Leishmania (L.). In this study, mRNA expression of Interleukin (IL)-12p40, IL-12p35, Interferon (IFN)-?, transforming growth factor (TGF)-?, IL-1, and IL-4 in PMN of volunteers stimulated with L. major promastigotes has been investigated by real-time PCR and the results were confirmed by flow cytometer. The results showed that L. major did not induce mRNA expression of IL12p40, IL12p35, IFN-?, and TGF-? in PMN, while IL-1 and IL-4 mRNA were induced. Flow cytometry results confirmed no IFN-? production by PMN with or without stimulation. IL-12p70 was present in untreated and L. major-treated PMN, and these cells release IL-12 following incubation with L. major. Significant amount of IL-1 even without treatment with promastigotes was detected in PMN. Moreover, the proportion of PMN, which produce IL-1 in response to L. major, was increased compared with the percent of unstimulated IL-1-producing PMN. The results showed the accumulation of small amounts of IL-4 in PMN after stimulation. In conclusion, our results indicate that IL-12 and IL-1 are pre-stored in human PMN, nor L. major induces IL-1 and IL-4, but not IL-12, IFN-?, nor TGF-? expression in these cells. PMID:24698213

Keyhani, Alireza; Riazi-Rad, Farhad; Pakzad, Saeed Reza; Ajdary, Soheila

2014-09-01

60

Peripheral blood leukocytes of cows with subclinical endometritis show an altered cellular composition and gene expression.  

PubMed

Subclinical endometritis (SCE) is an important postpartum disease in dairy cows, but conventional cytobrush diagnosis often gives imprecise results. The aim of this study was to analyze disease-associated changes in peripheral blood as potential diagnostic parameters. Cellular subpopulations of blood leukocytes from cows with or without SCE (45-55 days postpartum) were flow-cytometrically quantified. Gene expression of whole blood leukocytes was assessed by PAXgene analysis. Subclinical endometritis cows showed significantly higher number of blood mononuclear cells and neutrophils. Among mononuclear cells, numbers of B-cells, NK-cells, and CD172a-positive monocytes were significantly elevated. Compared with non-SCE cows, blood leukocytes of SCE cows significantly expressed higher copy numbers of CXCL8, TNF, and IL12. To test whether circulating plasma factors are responsible for these changes, leukocytes, polymorphonuclear cells, and monocyte subpopulations (classical, intermediate, nonclassical) of healthy cows were stimulated with plasma of SCE and non-SCE cows. Although gene expression of whole leukocytes and polymorphonuclear cells remained unaltered, plasma from SCE animals significantly elevated expressed messenger RNA copy numbers of CXCL8, CXCL1, and IL1B in intermediate monocytes. In conclusion, elevated number of selected mononuclear subpopulations in peripheral blood and enhanced expression of distinct genes encoding for inflammatory mediators in blood leukocytes reflect the subclinical uterine inflammatory process in cows. Whether the observed changes in the periphery of SCE cows are the consequence of the uterine inflammatory process, or whether they affect the pathogenesis of the disease is currently unknown. PMID:24560452

Düvel, Anna; Maaß, Janine; Heppelmann, Maike; Hussen, Jamal; Koy, Mirja; Piechotta, Marion; Sandra, Olivier; Smith, David G E; Sheldon, Iain Martin; Dieuzy-Labaye, Isabelle; Zieger, Peter; Schuberth, Hans Joachim

2014-04-15

61

Apoptosis and necrosis of blood and milk polymorphonuclear leukocytes  

E-print Network

milk somatic cell counts (SCC) are used as an indicator for bovine mastitis. Dur- ing mastitis-V (green) and propidium iodide (red). Using this technique three different subpopulations of bovine PMN green and high intensive red fluorescence) and viable cells (low intensive green and low intensive red

Paris-Sud XI, Université de

62

Stimulation of Polymorphonuclear Leukocyte Chemotaxis by Cold Precipitable Complexes Containing DNA-Anti-DNA in Active Nephritis of Systemic Lupus Erythematosus  

Microsoft Academic Search

DNA-anti-DNA cold precipitable complexes (CPC) isolated from 21 patients with systemic lupus erythematosus were tested for their ability to stimulate polymorphonuclear leukocyte (PMN) chemotaxis and to activate complement in vitro. The complexes activated complement in 20 out of 21 cases and stimulated PMN chemotaxis in 13 out of 21. The stimulation of chemotaxis was present in the majority of patients

A. Losito; C. Cecchini; L. Pittavini; I. Zampi

1982-01-01

63

Human Polymorphonuclear Leukocyte Chemotaxis as a Tool in Detecting Biological Early Effects in Workers Occupationally Exposed to Low Levels of n-HeXane  

Microsoft Academic Search

Human polymorphonuclear leukocytes (PMN) were chosen to measure two cellular end points- chemotaxis and respiratory burst - and to verify whether they could function as biomarkers of early effect in detecting occupational exposure to n-hexane of apparently healthy shoe workers, without any electroneuromyographic (ENMG) abnormality.Chemotaxis, but not respiratory burst, was found to be impaired. A negative linear correlation between chemotaxis

Mario Governa; Matteo Valentino; Isabella Visanà; Francesca Monaco

1994-01-01

64

Blood leukocyte and spleen lymphocyte immune response of spleen lymphocytes and whole blood leukocytes of hamsters  

SciTech Connect

This study was designed to evaluate the effects of chronic physical activity on the immune response of spleen lymphocytes and whole blood leukocytes of hamsters. Animals were kept sedentary or allowed to exercise spontaneously on running wheels for eight weeks. Physically active animals averaged 12 kilometers per day. The immune response of spleen lymphocytes whole blood leukocytes was evaluated by {sup 3}H-thymidine incorporation in response to Concanavalin A or lipopolysaccharide. There was no treatment effect between physically active and sedentary hamster in response of spleen lymphocytes. The immune response of whole blood leukocytes to these mitogens was significantly greater in physically active vs. sedentary hamsters. These results demonstrate that chronic physical activity has the capacity to modulate immunoresponses.

Peters, B.A.; Sothmann, M.; Wehrenberg, W.B. (Univ. of Wisconsin, Milwaukee (USA))

1989-01-01

65

Pasteurella haemolytica leukotoxin enhances production of leukotriene B4 and 5-hydroxyeicosatetraenoic acid by bovine polymorphonuclear leukocytes.  

PubMed Central

The influence of the leukotoxin of Pasteurella haemolytica on the generation of arachidonic acid metabolites by bovine polymorphonuclear leukocytes (PMNs) was investigated. PMNs released 5-, 12-, and 15-hydroxyeicosatetraenoic acids (5-, 12-, and 15-HETE) and leukotriene B4 (LTB4) upon stimulation with arachidonic acid. The leukotoxin preparations dose dependently enhanced the release of the 5-lipoxygenase products 5-HETE and LTB4 in arachidonic acid-stimulated PMNs, whereas the release of 12- and 15-HETE was not affected. The enhanced release of LTB4 and 5-HETE was not due to a decreased cellular retention of the 5-lipoxygenase products. In addition, leukotoxin preparations by themselves were also able to induce LTB4 and 5-HETE production in the absence of exogenous arachidonic acid. Generation of 5-lipoxygenase products by PMNs stimulated by leukotoxin may represent an important cellular event that occurs during infections with P. haemolytica. PMID:1322370

Henricks, P A; Binkhorst, G J; Drijver, A A; Nijkamp, F P

1992-01-01

66

The relationship between glycated hemoglobin and polymorphonuclear leukocyte leukotriene B4 release in people with diabetes mellitus.  

PubMed

In order to evaluate polymorphonuclear leukocyte (PMN) activity in diabetes mellitus, leukotriene B4 (LTB4) levels were measured in sixty patients, 31 affected with Type 1 diabetes mellitus and 29 affected with Type 2 diabetes mellitus. The LTB4 levels (12.1+/-0.2 pg/100 microl) in diabetic patients were higher compared to those of the control group (7.9+/-0.1 pg/100 microl) (P < 0.001), and remained significantly higher (P < 0.001) (12.8+/-0.2 pg/100 microl) than in the control group (11.0+/-0.2 pg/100 microl) after stimulation with calcium ionophore. A significant and positive correlation between glycated hemoglobin and LTB4 was demonstrated (P < 0.001, r = 0.80). This study demonstrates that in diabetic patients there is a PMN activation and that this activation is correlated to glycated hemoglobin level. PMID:10580615

Parlapiano, C; Danese, C; Marangi, M; Campana, E; Pantone, P; Giovanniello, T; Zavattaro, E; Sanguigni, S

1999-10-01

67

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

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.

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

1988-06-01

68

Carnosic acid and carnosol potently inhibit human 5-lipoxygenase and suppress pro-inflammatory responses of stimulated human polymorphonuclear leukocytes.  

PubMed

Carnosic acid (CA) and carnosol (CS) are phenolic diterpenes present in several labiate herbs like Rosmarinus officinalis (Rosemary) and Salvia officinalis (Sage). Extracts of these plants exhibit anti-inflammatory properties, but the underlying mechanisms are largely undefined. Recently, we found that CA and CS activate the peroxisome proliferator-activated receptor gamma, implying an anti-inflammatory potential on the level of gene regulation. Here we address short-term effects of CA and CS on typical functions of human polymorphonuclear leukocytes (PMNL). We found that (I), CA and CS inhibit the formation of pro-inflammatory leukotrienes in intact PMNL (IC(50)=15-20 microM [CA] and 7 microM [CS], respectively) as well as purified recombinant 5-lipoxygenase (EC number 1.13.11.34, IC(50)=1 microM [CA] and 0.1 microM [CS], respectively), (II) both CA and CS potently antagonise intracellular Ca(2+) mobilisation induced by a chemotactic stimulus, and (III) CA and CS attenuate formation of reactive oxygen species and the secretion of human leukocyte elastase (EC number 3.4.21.37). Together, our findings provide a pharmacological basis for the anti-inflammatory properties reported for CS- and CA-containing extracts. PMID:18508031

Poeckel, Daniel; Greiner, Christine; Verhoff, Moritz; Rau, Oliver; Tausch, Lars; Hörnig, Christina; Steinhilber, Dieter; Schubert-Zsilavecz, Manfred; Werz, Oliver

2008-07-01

69

Prevention of stromal ulceration in the alkali-burned rabbit cornea by glued-on contact lens. Evidence for the role of polymorphonuclear leukocytes in collagen degradation  

Microsoft Academic Search

Stromal ulceration of the alkali-burned rabbit cornea ivas found to be associated invariably with phagocytically active polymorphonuclear leukocytes (PMNs). A glued-on methylmethacry- late lens applied to corneas soon after burning, however, prevented re-epithelialization and also prevented PMN infiltration of the stroma and stromal ulceration. Subsequent partial detachment or complete removal of the lens residted in epithelial resurfacing of the stroma,

K. R. Kenyon; M. Berman

1979-01-01

70

An Endotoxin-Induced Factor Distinct From Interleukin1 and Tumour Necrosis Factor a Produced by the THP1 Human Macrophage Line Stimulates Polymorphonuclear Leukocyte Infiltration In Vivo  

Microsoft Academic Search

Endotoxin and gram-negative bacteria induce vigorous inflammatory reactions. Our pre- vious work showed that rabbit macrophages (Mo) incubated with endotoxin produce a 45,000 dalton protein that recruited polymorphonuclear leukocytes (PMNL) into the skin of rabbits. This factor was separated from interleukln-i (IL-i) but could not be unequiv- ocally distinguished from rabbit tumour necrosis factor (TNFa). Here we have examined the

Pal Megyeri; Thomas B. Issekutz; Andrew C. lssekutz

71

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

SciTech Connect

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.

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

1989-01-01

72

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

PubMed

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

Van Gennip, Maria; Christensen, Louise Dahl; Alhede, Morten; Phipps, Richard; Jensen, Peter Østrup; Christophersen, Lars; Pamp, Sünje Johanna; Moser, Claus; Mikkelsen, Per Jensen; Koh, Andrew Y; Tolker-Nielsen, Tim; Pier, Gerald B; Høiby, Niels; Givskov, Michael; Bjarnsholt, Thomas

2009-07-01

73

Purification of the active C5a receptor from human polymorphonuclear leukocytes as a receptor - G sub i complex  

SciTech Connect

The authors have isolated, in an active state, the C5a receptor from human polymorphonuclear leukocytes. The purification was achieved in a single step using a C5a affinity column in which the C5a molecule was coupled to the resin through its N terminus. The purified receptor, like the crude solubilized molecule, exhibited a single class of high-affinity binding sites with a K{sub d} of 30 pM. Further, the binding of C5a retained its sensitivity to guanine nucleotides, implying that the purified receptor contained a guanine nucleotide-binding protein (G protein). SDS/PAGE revealed the presence of three polypeptides with molecular masses of 42, 40, and 36 kDa, which were determined to be the C5a-binding subunit and the {alpha} and {beta} subunits of G{sub i}, respectively. The 36- and 40-kDa polypeptides were identified by immunoblotting and by the ability of pertussis toxin to ADP-ribosylate the 40-kDa molecule. These results confirm their earlier hypothesis that the receptor exists as a complex with a G protein in the presence or absence of C5a. The tight coupling between the receptor and G protein should make possible the identification of the G protein(s) involved in the transduction pathways used by C5a to produce its many biological effects.

Rollins, T.E.; Siciliano, S.; Kobayashi, S.; Cianciarulo, D.N.; Bonilla-Argudo, V.; Collier, K.; Springer, M.S. (Merck Sharp and Dohme Research Lab., Rahway, NJ (United States))

1991-02-01

74

C5a-induced hemodynamic and hematologic changes in the rabbit. Role of cyclooxygenase products and polymorphonuclear leukocytes.  

PubMed Central

Hemodynamic and hematologic changes occurring after intravascular complement activation have implicated the anaphylatoxins in this response. In this study, the hemodynamic and hematologic effects of purified C5a were investigated in rabbits; and involvement of prostanoids, histamine, and polymorphonuclear leukocytes (PMNs) were examined. The anaphylatoxin C5a induces a reversible systemic arterial hypotension which coincides with an increase in central venous pressure (CVP), decreased cardiac output (CO), increased plasma prostanoid levels, as well as neutropenia. Total peripheral resistance (TPR) remained unchanged. The cyclooxygenase inhibitor indomethacin abolished the C5a-induced hypotension and normalized plasma prostanoid levels without altering the C5a-induced neutropenia. The thromboxane (Tx) A2 synthetase inhibitor dazoxiben reduced TxB2 plasma levels and increased 6-keto-prostaglandin PGF1 alpha and PGE2 levels without altering the hypotensive response. However, with dazoxiben treatment both TPR and CVP decreased. The H2-receptor antagonist cimetidine reduced C5a-induced hypotension and diminished prostanoid release. Both the hypotensive response and elevated prostanoid release were observed after C5a challenge in animals rendered neutropenic prior to challenge. It is concluded that C5a-induced arterial hypotension in the rabbit is a PMN-independent reaction, mediated through cyclooxygenase products and, to some degree, by histamine. The mechanism producing systemic arterial hypotension does not seem to involve peripheral vasodilation but appears to be a secondary effect of pulmonary vasoconstriction, possibly mediated by TxA2. PMID:3115110

Lundberg, C.; Marceau, F.; Hugli, T. E.

1987-01-01

75

Anti-C5a monoclonal antibodies and pulmonary polymorphonuclear leukocyte infiltration--endothelial dysfunction by venous gas embolism.  

PubMed

Venous gas embolism (VGE) impairs endothelial function although there is no apparent mechanical damage to the endothelial layer. We investigated whether a monoclonal antibody against the complement anaphylatoxine C5a would affect endothelial dysfunction and pulmonary polymorphonuclear leukocyte infiltration caused by low-grade VGE. Six rabbits were pre-treated with the anti-C5a monoclonal antibody whereas a sham monoclonal antibody was administrated to six other animals 30 min before VGE. Six untreated rabbits subjected to an identical protocol except antibody treatment were used for control. The monoclonal anti-C5a antibody reduced PMN infiltration compared to the control group ( P<0.03). There were no major signs of apoptosis in endothelial cells inside the pulmonary artery in any of the examined animals. There was reduced PMN infiltration and improved endothelium-dependent relaxation in the sham-antibody group, these effects were however not significant. In conclusion, anti-C5a protects the endothelium against injury caused by small amounts of gas bubbles. PMID:12682835

Nossum, Vibeke; Hjelde, Astrid; Bergh, Kåre; Ustad, Anne-Lise; Brubakk, Alf O

2003-05-01

76

Role of the Yersinia pestis Ail protein in preventing a protective polymorphonuclear leukocyte response during bubonic plague.  

PubMed

The ability of Yersinia pestis to forestall the mammalian innate immune response is a fundamental aspect of plague pathogenesis. In this study, we examined the effect of Ail, a 17-kDa outer membrane protein that protects Y. pestis against complement-mediated lysis, on bubonic plague pathogenesis in mice and rats. The Y. pestis ail mutant was attenuated for virulence in both rodent models. The attenuation was greater in rats than in mice, which correlates with the ability of normal rat serum, but not mouse serum, to kill ail-negative Y. pestis in vitro. Intradermal infection with the ail mutant resulted in an atypical, subacute form of bubonic plague associated with extensive recruitment of polymorphonuclear leukocytes (PMN or neutrophils) to the site of infection in the draining lymph node and the formation of large purulent abscesses that contained the bacteria. Systemic spread and mortality were greatly attenuated, however, and a productive adaptive immune response was generated after high-dose challenge, as evidenced by high serum antibody levels against Y. pestis F1 antigen. The Y. pestis Ail protein is an important bubonic plague virulence factor that inhibits the innate immune response, in particular the recruitment of a protective PMN response to the infected lymph node. PMID:21969002

Hinnebusch, B Joseph; Jarrett, Clayton O; Callison, Julie A; Gardner, Donald; Buchanan, Susan K; Plano, Gregory V

2011-12-01

77

Enhanced oxidative burst in immunologically activated but not elicited polymorphonuclear leukocytes correlates with fungicidal activity.  

PubMed

Polymorphonuclear neutrophils (PMN) induced locally in immune mice by intraperitoneal injection of antigen exhibit enhanced fungicidal activity compared with PMN elicited with thioglycolate. The mechanism of the differences in these PMN populations was studied. Sublethal infection was used to produce immunity to Blastomyces dermatitidis. A correlation was sought between the ability of PMN to kill, or not kill, B. dermatitidis and the production of the oxidative burst, as measured by luminol-enhanced chemiluminescence (CL). Although elicited PMN cocultured with Candida albicans produced a burst of CL and were candidacidal, killing did not occur when PMN were cocultured with B. dermatitidis. Lack of killing of B. dermatitidis by elicited PMN correlated with lack of stimulation of a brisk oxidative burst. In contrast to elicited PMN, PMN induced by B. dermatitidis antigen responded to this fungus with a burst of CL and a significant reduction of inoculum CFU (80%). Furthermore, these PMN when cocultured with C. albicans produced an enhanced burst of CL, and killing was enhanced compared with that by elicited PMN, e.g., 86 versus 58%. The CL burst and killing of B. dermatitidis by antigen-induced PMN was abrogated in the presence of catalase, implying a critical role for hydrogen peroxide. Partial but significant depression of CL and killing in the presence of dimethyl sulfoxide, a hydroxyl radical scavenger, identified hydroxyl radical, or its metabolites, as a toxic product(s) responsible for a significant fraction of fungicidal activity. These results indicate that the metabolic activity and microbicidal activity of PMN can be altered (enhanced) at the site of an immunological reaction and thus could constitute an important factor in resistance. PMID:3894234

Brummer, E; Sugar, A M; Stevens, D A

1985-08-01

78

Leukocyte flotation during gravity sedimentation of the whole blood.  

PubMed

The original Westergren blood sedimentation technique was modified to assess leukocyte sedimentation properties. The relative change of leukocyte and erythrocyte counts was measured in the upper half section of blood column in vertically positioned sedimentation tubes in 10-minute-intervals for 60 minutes. During the first 20 minutes of gravity sedimentation, the leukocytes taken from critically ill patients showed upward flotation, however, healthy individuals' leukocytes demonstrated slight sedimentation. The upward flotation rate of leukocytes seemed less dependent on erythrocyte sedimentation during the first 15 minutes of sedimentation time than after it. Based on this observation, the sedimentation properties of leukocytes were characterized by the leukocyte antisedimentation rate taken at the 15th minute of sedimentation time (LAR15). Erythrocyte aggregability index, plasma fibrinogen concentration and native leukocyte count did not correlate to LAR15 in healthy volunteers (n = 25). However, LAR15 was correlated to leukocyte adherence (p < 0.01), to whole blood viscosity (p < 0.05), to hematocrit (p < 0.05) and to the conventional erythrocyte sedimentation rate (p < 0.05). PMID:10711819

Bogar, L; Tekeres, M

2000-01-01

79

Cytokine production by leukocytes of Papillon-Lefèvre syndrome patients in whole blood cultures.  

PubMed

Papillon-Lefèvre syndrome (PLS) is characterised by aggressively progressive periodontitis combined with palmo-plantar hyperkeratosis. It is caused by "loss of function" mutations in the cathepsin C gene. The hypothesis behind this study is that PLS patients' polymorphonuclear leukocytes (PMNs) produce more proinflammatory cytokines to compensate for their reduced capacity to neutralize leukotoxin and to eliminate Aggregatibacter actinomycetemcomitans. Production of more interleukin (IL)-8 would result in the attraction of more PMNs. The aim of this study was to evaluate the cytokine profile in PLS patients' blood cultures. Blood was sampled from eight PLS patients (one female) from six families (antiinfective therapy completed: six; edentulous: two) with confirmed cathepsin C mutations and deficient enzyme activity. Nine healthy males served as controls. Whole blood cultures were stimulated with highly pure lipopolysaccharide (LPS) from Escherichia coli R515 and IL-1? plus tumor necrosis factor (TNF)-?. Thereafter, release of IL-1? (stimulation: LPS and LPS plus adenosine triphosphate), IL-6, IL-8, interferon-inducible protein (IP)-10, and interferon (IFN)-? (stimulation: LPS, IL-1?/TNF?) were detected by ELISA. Medians of cytokine release were, with the exception of IP-10, slightly higher for PLS than for controls' cultures. None of these differences reached statistical significance. Increased production of IL-1?, IL-6, IL-8, IP-10, or IFN? as a significant means to compensate for diminished activity and stability of polymorphonuclear leukocyte-derived proteases could not be confirmed in this study. Cytokine profiles in blood cultures may not be used to identify PLS patients. PMID:21380503

Sadik, Christian D; Noack, Barbara; Schacher, Beate; Pfeilschifter, Josef; Mühl, Heiko; Eickholz, Peter

2012-04-01

80

Thyroid hormone and thyrotropin regulate intracellular free calcium concentrations in human polymorphonuclear leukocytes: in vivo and in vitro studies.  

PubMed

Intracellular free calcium concentrations (Ca++i) were studied in polymorphonuclear leukocytes (PMNs) from 13 athyreotic patients who had been previously treated by total thyroidectomy and radioiodine therapy for differentiated thyroid carcinoma, and from age- and sex-matched euthyroid healthy controls. Patients were studied twice, when hypothyroid (visit 1) and after restoration of euthyroidism by L-T4 TSH-suppressive therapy (visit 2). PMNs from patients at visit 1 had significantly lower resting (Ca++)i levels compared to both visit 2 and controls. Values at visit 2 did not differ from those of the controls. Stimulus-induced (Ca++)i rise was also significantly blunted at visit 1 and normalized at visit 2, possibly through a differential contribution of distinct intracellular Ca++ stores, as suggested by the response pattern to the chemotactic agent, N-formyl-Met-Leu-Phe (fMLP), to the selective SERCA pump inhibitor, thapsigargine, and to the mitochondrial uncoupler, carbonyl cyanide p-trifluoromethoxyphenyl-hydrazone (FCCP). In vitro treatment of PMNs from healthy subjects with high TSH concentrations impaired intracellular Ca++ store function. Both resting (Ca++)i levels and fMLP-induced (Ca++)i rise increased in the presence either of low-concentration TSH or of T4, but effects of TSH and T4 were not additive. T3, rT3, and TRIAC had no effect. In conclusion, this study provides evidence for a direct relationship between thyroid status and (Ca++)i homeostasis in human PMNs, mainly related to direct actions of TSH and T4 on these cells. PMID:16569353

Marino, F; Guasti, L; Cosentino, M; De Piazza, D; Simoni, C; Bianchi, V; Piantanida, E; Saporiti, F; Cimpanelli, M G; Crespi, C; Vanoli, P; De Palma, D; Klersy, C; Frigo, G M; Bartalena, L; Venco, A; Lecchini, S

2006-01-01

81

Clinically effective monoclonal antibody 3F8 mediates nonoxidative lysis of human neuroectodermal tumor cells by polymorphonuclear leukocytes.  

PubMed

Most studies of antibody-dependent cellular cytotoxicity (ADCC) by polymorphonuclear leukocytes (PMN) have supported oxidative lytic processes. This may be because the studies used nonhuman or nonneoplastic cells that were highly sensitive to reactive oxygen species or were small enough to be phagocytosed by PMN. We therefore investigated whether oxygen radicals participate in PMN cytotoxicity toward human neuroectodermal solid tumor cells sensitized by 3F8, which is an anti-ganglioside GD2 murine IgG3 monoclonal antibody with documented anticancer activity in humans. A 4-h 51Cr release assay was used to assess tumor cell lysis by hydrogen peroxide, superoxide, and hypochlorite. Nine of 11 GD2(+) human melanoma and neuroblastoma cell lines had equal or greater resistance to these oxidants as compared to a GD2(-) human carcinoma line (SKBr1-III) found by others (and confirmed by us) to be significantly more resistant to oxidative lysis than a murine cell line (P388D1) representative of those commonly used in cytotoxicity assays. To facilitate detection of oxidant-mediated lysis, subsequent studies of 3F8-mediated ADCC used GD2(+) targets that were relatively sensitive and others that were relatively resistant to oxygen radicals. Normal PMN and PMN obtained from children with chronic granulomatous disease, which do not generate reactive oxygen species, were equally effective in ADCC. Granulocyte-macrophage colony-stimulating factor, which primes oxidative responses of normal but not of chronic granulomatous disease PMN, enhanced ADCC by both kinds of PMN. During ADCC of 3F8-sensitized targets, with or without granulocyte-macrophage colony-stimulating factor, GD2(-) "innocent bystander" tumor cells (including P388D1) were not lysed, a finding consistent with unimportant extracellular release of cytotoxic mediators. Finally, antioxidant and antimyeloperoxidase moieties did not block ADCC. We conclude that oxidants are not key factors in 3F8-mediated lysis by PMN of human neuroectodermal tumor cells. PMID:1654202

Kushner, B H; Cheung, N K

1991-09-15

82

Phagocytic Activity and State of Bactericidal Systems in Polymorphonuclear Leukocytes from Patients with Alzheimer's Disease  

Microsoft Academic Search

Phagocytic activity of peripheral blood neutrophils underwent phase changes in patients with Alzheimer's disease. Neutrophils retained the ability to engulf microbes, but their digestive activity decreased at the early stage of Alzheimer's disease. At the late stage we observed a decrease in the count of phagocytizing neutrophils, reduction of myeloperoxidase activity, and increase in the content of cationic proteins.

T. V. Davydova; V. G. Fomina; N. I. Voskresenskaya; O. A. Doronina

2003-01-01

83

Role of neutrophil polymorphonuclear leukocytes during bovine coliform mastitis: physiology or pathology?  

PubMed

The review compiles some major findings concerning the inflammatory reaction in the mammary gland of dairy cows within the physiological context of the lactation cycle. The dual role of the PMN leukocyte in defense and tissue damage during experimentally induced coliform mastitis, especially around parturition and during early lactation, is highlighted. This disease affects many high producing cows in dairy herds and may cause several cases of death per year in the most severe cases. Most researchers now accept that the PMN is a key factor in the cows' defense against intramammary infection with E coli. During diapedesis of PMN into the mammary gland, several functionally important receptors are up-regulated, allowing for a more efficient phagocytosis and killing of invading pathogens. While PMN are phagocytosing and destroying the invading pathogens, they inadvertently release chemical mediators which induces swelling of secretory epithelium cytoplasm, sloughing of secretory cells, and decreased secretory activity. Permanent scarring will result in a loss of milk production. PMN's act as friends and as foes and are important components in the balance between mammary defense and damage. The mammary gland is a complex open self-regulatory system with a continuous flow of matter, energy and information. Metabolically, it has absolute priority over many other tissues except the brain. Self-regulation with change over time is characterized by a dynamic equilibrium between two mechanisms: homeostatic and homeorhetic. The defense against invaders by innate immunity and auto-repair of the damaged tissues are covered by homeostatic mechanisms while colostrogenesis and maintenance of milk secretion are controlled by mainly homeorhetic mechanisms. However, also innate immunity has to function and develop in time, depending on the lactation cycle, and its behavior and evolution in time in such a dynamical system is a challenge and a problem at the same time. In such a complex dynamic situation it is not surprising that physiology is not far away from pathology. E. coli mastitis can be a severe problem during the beginning of lactation whereas it is completely self-curing after peak lactation (8 weeks). The approach to focus on the PMN doesn't mean that the defense of the mammary gland is more simple than in other tissues. The defense of mammary gland is characterized by its complexity and over the last years many data show that there are tight connections with the mononuclear cells in mammary gland tissue. Today it is known that T cells play a central role in orchestrating the immune response. However, because of the peculiar interest in the PMN of the authors during the last 10 years, the immunobiology of the mononuclear cells in the mammary gland is not covered. PMID:15074078

Burvenich, C; Monfardini, E; Mehrzad, J; Capuco, A V; Paape, M J

2004-01-01

84

Decoration of Histophilus somni lipooligosaccharide with N-acetyl-5-neuraminic acid enhances bacterial binding of complement factor H and resistance to killing by serum and polymorphonuclear leukocytes.  

PubMed

The incorporation of N-acetyl-5-neuraminic acid (Neu5Ac), or sialic acid, onto surface components of some bacterial species may enhance their virulence. We have previously shown that Neu5Ac can be incorporated onto the lipooligosaccharide (LOS) of the bovine pathogen Histophilus somni, resulting in diminished antibody binding and enhanced serum resistance (Inzana et al., 2002. Infect. Immun. 70, 4870). In the present study, we assessed the effect of sialylation of H. somni LOS on the interaction with bovine innate host defenses. Incubation of non-sialylated H. somni with pre-colostral calf serum (PCS) resulted in dose-dependent, complement-mediated killing of the bacteria by the alternative pathway. However, sialylated H. somni was significantly more resistant to killing at any of the concentrations of PCS used. Sialylated H. somni LOS activated and consumed less complement than non-sialylated LOS, as determined by reduction in hemolysis of opsonized red blood cells, and by Western blotting of C(3) activation products. Sialylated H. somni bound more factor H and iC(3)b and less C(3) than non-sialylated bacteria, as determined by enzyme-linked immunosorbent assay, supporting the deficiencies observed in complement activation and consumption by sialylated LOS. Sialylation of H. somni LOS inhibited both polymorphonuclear leukocyte phagocytosis of (3)H-thymidine-labeled bacteria and intracellular killing of the bacteria, compared to non-sialylated bacteria. Furthermore, sialylated H. somni bound less non-specific antibodies in normal bovine sera than non-sialylated bacteria. Therefore, sialylation of H. somni LOS had profound effects on resistance of the bacteria to innate bovine host defenses, which should be taken into consideration during in vitro studies of H. somni. PMID:22868182

Inzana, Thomas J; Balyan, Rajiv; Howard, Michael D

2012-12-28

85

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

Federal Register 2010, 2011, 2012, 2013

...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''...

2011-01-31

86

Effect of etizolam (Depas) on production of superoxide anion by platelet-activating factor and N-formyl-methionyl-leucyl-phenylalanine-stimulated guinea pig polymorphonuclear leukocytes.  

PubMed

Effect of etizolam on platelet activating factor (PAF) and N-formyl-methionyl-leucyl-phenylalanine (FMLP)-induced superoxide anion (O2-) production in guinea pig polymorphonuclear leukocytes (PMNL) was investigated. Etizolam showed the inhibitory effect on PAF-induced O2- production concentration dependently, with an IC50 value of 4.7 microM, but it had no inhibitory effect on FMLP-induced O2- production at 100 microM. These results suggest that etizolam has a selectively strong inhibitory effect on PAF-induced O2- production in guinea pig PMNL. PMID:2848961

Aratani, H; Nishida, Y; Terasawa, M; Maruyama, Y

1988-06-01

87

Modulation of polymorphonuclear leukocyte microbicidal activity and oxidative metabolism by fibrinogen degradation products D and E.  

PubMed Central

Fibrinogen degradation products (FDP) D and E are typically present in blood of patients with disseminated intravascular coagulation and related conditions in which granulocyte (PMN) defense against bacterial infection may be compromised. This study was intended to determine whether FDP modify PMN functions critical to their bactericidal activity. Incubation of human PMN and Escherichia coli with 50-100 micrograms/ml FDP did not affect phagocytosis, but reduced by greater than 90% the cells' ability to inhibit bacterial colony growth compared with control PMN incubated with albumin or fibrinogen. FDP (10-100 micrograms/ml) inhibited PMN O2- release and chemotaxis stimulated by FMLP by 17-50% (P less than 0.005) and 41% (P less than 0.01), respectively. Fragment E3, and not fragment D1, was primarily responsible for inhibition of FMLP-induced PMN O2- release. Phorbol myristate acetate (10 ng/ml), 1-oleoyl-2-acetylglycerol (10(-6) M), AA (4.2 x 10(-5) M), and zymosan-activated serum-stimulated PMN O2- release were also decreased 37-63% by FDP compared with control protein. There are at least two mechanisms by which FDP may impair PMN responses. With respect to FMLP, FDP (16-100 micrograms/ml) inhibited specific binding to the cell surface over a ligand concentration range of 1.4-85 nM [3H]FMLP. In contrast, FDP did not effect the extent of phorbol ester binding to PMN but blocked activation of protein kinase C. These data suggest that elevated plasma FDP inhibit several PMN functions critical to the bactericidal role of these inflammatory cells. Images PMID:2542377

Kazura, J W; Wenger, J D; Salata, R A; Budzynski, A Z; Goldsmith, G H

1989-01-01

88

Oxidative Stress in Leukocytes Is a Possible Link Between Blood Pressure, Blood Glucose, and C-Reacting Protein  

Microsoft Academic Search

Because oxidative stress and inflammation are believed to play roles in the pathogenesis of cardiovascular diseases, oxidative stress in polymorphonuclear leukocytes (PMNs) and mononuclear cells (MNCs) has been measured. A total of 529 subjects participated this study. Intracellular oxidative stress in PMNs and MNCs was measured by gated flow cytometry using carboxyfluorescin diacetate bis-acetoxymethyl ester. C-reacting protein (CRP), insulin action

Kenichi Yasunari; Kensaku Maeda; Munehiro Nakamura; Junichi Yoshikawa

89

Dielectric Characterization of Leukocytes from Human Blood  

Microsoft Academic Search

The impedance spectroscopy technique was used to perform a comparative study on blood samples from leukemia and from healthy people. An electric circuit model was used to determine the numerical parameters of each sample. The capacitive properties, reflected in the constant phase element of a Cole model, show the capability to analyze the dielectric and spectral behavior of blood cells,

Jesús Bernal-Alvarado; Gilberto Gutiérrez; Antonio Sánchez; Modesto Sosa; Francisco Hernández; Carlos Guerrero; Julio C. Villagómez; Pascual Palomares; Ana L. Contreras

2006-01-01

90

Dielectric Characterization of Leukocytes from Human Blood  

NASA Astrophysics Data System (ADS)

The impedance spectroscopy technique was used to perform a comparative study on blood samples from leukemia and from healthy people. An electric circuit model was used to determine the numerical parameters of each sample. The capacitive properties, reflected in the constant phase element of a Cole model, show the capability to analyze the dielectric and spectral behavior of blood cells, in particular, with cells from leukemia. The spectra and the capacitive parameters are shown, as well as de model fitted to the experimental data.

Bernal-Alvarado, Jesús; Gutiérrez, Gilberto; Sánchez, Antonio; Sosa, Modesto; Hernández, Francisco; Guerrero, Carlos; Villagómez, Julio C.; Palomares, Pascual; Contreras, Ana L.

2006-09-01

91

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

Federal Register 2010, 2011, 2012, 2013

...2001D-0037)] Guidance for Industry: Pre-Storage Leukocyte Reduction of Whole Blood and...entitled ``Guidance for Industry: Pre- Storage Leukocyte Reduction of Whole Blood and...establishments with recommendations for pre-storage leukocyte reduction of Whole Blood...

2012-09-25

92

Relationship between zinc malnutrition and alterations in murine peripheral blood leukocytes  

SciTech Connect

Studies using a murine model have shown that the immune system responds rapidly and adversely to zinc deficiency. The extent of alteration of peripheral blood leukocytes (PBL) and immunoglobulin levels were investigated in four zinc dietary groups: zinc adequate (ZA); restricted fed zinc adequate (RZA); marginal zinc deficient (MZD, 72-76% of ZA mouse weight); and severely zinc deficient. The peripheral white blood cell count was 3.66 {plus minus} 1.08 {times} 10{sup 6} cells/ml for ZA mice decreasing by 21%, 28% and 54% for RZA, MZD and SZD mice respectively. An equally dramatic change in the flow cytometric light scatter profile was found. ZA mice had 66% lymphocytes and 21% polymorphonuclear granulocytes (PMN) in their peripheral blood while MZD and SZD mice contained 43% and 30% lymphocytes and 40% and 60% PMNs respectively. Analysis of the phenotypic distribution of specific classes of lymphocytes revealed ZA blood contained 25% B-cells and 40% T-cells (CD5{sup +}). B-cells decreased 40-50% for RZA and MZD mice and 60-70% for SZD mice. The decline in CD5{sup +} T-cells was more modest at 30% and 45% for MZD and SZD mice. A nearly 40% decline in both T{sub h} and T{sub c/s} cells was noted for both MZD and SZD mice. Radioimmunoassay of serum for changes in IgM and IgG content revealed no change among dietary groups while serum zinc decreased 10% for RZA mice and 50% for both MZD and SZD mice. The authors conclude that peripheral blood differential counts in concert with total B and T-cell phenotype may serve as indicators of zinc status while serum zinc and Ig will not.

King, L.E.; Morford, L.A.; Fraker, P.J. (Michigan State Univ., East Lansing (United States))

1991-03-15

93

Mononuclear and Polymorphonuclear Leukocyte Dispositions of Clarithromycin and Azithromycin in AIDS Patients Requiring Mycobacterium avium Complex Prophylaxis  

Microsoft Academic Search

The intracellular dispositions of clarithromycin and azithromycin in AIDS patients requiring Mycobacterium avium complex (MAC) prophylaxis were studied. The dispositions of both drugs in mononuclear and poly- morphonuclear leukocytes were markedly different. Our data support the proven efficacy of these agents for MAC prophylaxis since clarithromycin and azithromycin displayed sustained intracellular concentrations which exceeded their MICs for MAC throughout the

KHANH Q. BUI; JOCAROL MCNABB; CHONGHUA LI; CHARLES H. NIGHTINGALE; DAVID P. NICOLAU

1999-01-01

94

[Luminol-enhanced chemiluminescence of rabbit polymorphonuclear leukocytes: the nature of oxidants that directly induce luminol oxidation].  

PubMed

The present work deals with the reaction pathways, including the formation of hydroxyl radicals and chloroamines, which lead to luminol chemiluminescence caused by hypochlorite generation in a suspension of stimulated rabbit polymorphnonuclear leukocyte. Luminol-enhanced (0.02 mM) chemiluminescence of leukocytes stimulated by phorbol 12-myristate 13-acetate does not change in the presence of dimethyl sulfoxide at moderate concentrations (0.02-2.6 mM) at which it must show the specific ability to scavenge hydroxyl radicals. It suggests that no generation of hydroxyl radical with the participation of hypochlorite and superoxide anion takes place after the stimulation of polymorphnonuclear leukocytes. A high dimethyl sulfoxide concentrations (260 mM) a significant fall in chemiluminescence intensity, due to direct interaction of the scavenger with hypochlorite, is observed. Chemiluminescence intensity rose if luminol was added to a leukocyte suspension preliminary stimulated for 10 min. The effect results from the accumulation of hydrogen peroxide but not chloroamines. Exogenic amino acids and taurin at high concentrations (3-15 mM) weaken the chemiluminescence. The data obtained suggest that chemiluminescence in the system studied results predominantly from the direct initial reaction of hypochlorite with luminol. The chemiluminescence intensity is enhanced by hydrogen peroxide via the oxidation of luminol oxidation products. PMID:16521559

Roshchupkin, D I; Belakina, N S; Murina, M A

2006-01-01

95

Relationship between light scattering in flow cytometry and changes in shape, volume, and actin polymerization in human polymorphonuclear leukocytes  

Microsoft Academic Search

Using two different cytometers, an Epics Profile II and a FACScan, we determined the extent to which changes in forward and right angle scatter are a reliable measure for changes in polymorphonu- clear leukocyte (PMN) shape, volume, and actm po- lymerization and whether distinct types of shape changes in PMNs can be recognized. PMN stimulation can substantially change the positions

H. U. Keller; A. Fedier; R. Rohnert

96

Characterization of the interaction between recombinant human interferon-gamma and its receptor on human polymorphonuclear leukocytes  

SciTech Connect

The interaction of human recombinant interferon-gamma (rIFN-gamma) with human polymorphonuclear cells (PMN) was investigated. Bolton-Hunter radioiodinated rIFN-gamma bound to PMN in a specific and saturable manner. Eleven hundred binding sites were observed with a Ka of 0.56 x 10(10) M-1. Binding to PMN was rapid with a K1 of 9 x 10(5) M-1 sec-1 at 4{degree}C. At 37{degree}C binding was complete within 6 min. About 50% of bound ligand was internalized within 30 min at 37{degree}C. The receptor demonstrated moderate lability at 37{degree}C in culture. After 1 h at 37{degree}C, PMN lost 80% of their {sup 125}I-rIFN-gamma binding sites. This loss was reversed in part by the presence of interleukin-1 in the culture, but not tumor necrosis factor. These studies provide a framework for further investigation into the signalling process of rIFN-gamma on PMN.

Hansen, B.D.; Finbloom, D.S. (Walter Reed Army Institute of Research, Washington, DC (USA))

1990-01-01

97

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

PubMed Central

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

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

98

The role of diacylglyceride generation by phospholipase D and phosphatidic acid phosphatase in the activation of 5-lipoxygenase in polymorphonuclear leukocytes.  

PubMed

Diacylglycerides (DAGs) such as 1-oleoyl-2-acetyl-sn-glycerol (OAG) stimulate 5-lipoxygenase (5-LO) enzyme activity and function as agonists for human polymorphonuclear leukocytes (PMNL) to induce 5-LO product synthesis. Here, we addressed the role of endogenous DAG generation in agonist-induced 5-LO activation in human PMNL. Preincubation of PMNL with the phospholipase D (PLD) inhibitor 1-butanol potently suppressed 5-LO product synthesis induced by the Ca(2)(+) ionophore A23187 or thapsigargin (TG) and blocked A23187-evoked translocation of 5-LO from the cytosol to the nuclear membrane, analyzed by subcellular fractionation as well as by indirect immunofluorescence microscopy. Tertiary-butanol, a rather poor inhibitor of PLD, caused only moderate suppression of 5-LO and hardly inhibited 5-LO translocation. Interestingly, 1-butanol failed to inhibit 5-LO product formation when PMNL were stimulated with OAG (30 microM). Moreover, coincubation of A23187- or TG-stimulated PMNL with OAG reversed inhibition of 5-LO product formation by 1-butanol in a concentration-dependent manner (EC(50), approximately 1 muM) and also restored 5-LO translocation. In addition, inhibition of phosphatidic acid phosphatase (PA-P) by propranolol or bromoenol lactone caused suppression of 5-LO product formation and of translocation, which could be reversed by addition of exogenous OAG. Together, our data suggest that in agonist-stimulated PMNL, the endogenous formation of DAGs via the PLD/PA-P pathway determines 5-LO activation. PMID:18218859

Albert, Dana; Pergola, Carlo; Koeberle, Andreas; Dodt, Gabriele; Steinhilber, Dieter; Werz, Oliver

2008-04-01

99

Inhibition of apoptosis induced by heat shock preconditioning is associated with decreased phagocytosis in human polymorphonuclear leukocytes through inhibition of Rac and Cdc42.  

PubMed

The functionality of polymorphonuclear leukocytes (PMNL) and the exact process of the protective program employed by these cells in response to the heat shock (HS) remain ill-defined and debated. Particularly, the mechanism of phagocytic impairment induced by the HS and the molecular events associated with the delay of apoptosis used by these cells in such condition have given conflictual data. The aim of the present work is to study the consequences of the HS in different pathways involved in human PMNL apoptosis and subsequently in human PMNL phagocytic function. We demonstrated that HS (41 degrees C, 1 h) preconditioning induced inhibition of spontaneous PMNL apoptosis observed at 18 h in control cells incubated at 37 degrees C. This inhibition was characterized by absence of morphological nuclear changes, decrease of DNA fragmentation, low level of annexin V expression and decrease of caspase-3 activity. In parallel, HS increased both Hsp70 and Mcl-1 protein levels in PMNL. Phagocytosis of latex beads by PMNL was inhibited by HS (41 degrees C, 1 h) preconditioning despite an upregulation of CD11b, CD16 and CD47. Moreover, HS induced prolonged F actin depolymerization and inhibited both Rac and Cdc42 activation in PMNL. Finally, our results identify a new function of Mcl-1 in HS protection against apoptosis. PMID:17228324

Selva, Eric; Brest, Patrick; Loubat, Agnès; Lassalle, Sandra; Auberger, Patrick; Hofman, Paul

2007-01-01

100

Inhibitors of actin polymerisation stimulate arachidonic acid release and 5-lipoxygenase activation by upregulation of Ca2+ mobilisation in polymorphonuclear leukocytes involving Src family kinases.  

PubMed

Here, we show that actin polymerisation inhibitors such as latrunculin B (LB), and to a minor extent also cytochalasin D (Cyt D), enhance the release of arachidonic acid (AA) as well as nuclear translocation of 5-lipoxygenase (5-LO) and 5-LO product synthesis in human polymorphonuclear leukocytes (PMNL), challenged with thapsigargin (TG) or N-formyl-methionyl-leucyl-phenylalanine. The concentration-dependent effects of LB (EC50 approximately 200 nM) declined with prolonged preincubation (>3 min) prior TG and were barely detectable when PMNL were stimulated with Ca2+-ionophores. Investigation of the stimulatory mechanisms revealed that LB (or Cyt D) elicits Ca2+ mobilisation and potentiates stimulus-induced elevation of intracellular Ca2+, regardless of the nature of the stimulus. LB caused rapid but only moderate activation of p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinase (ERK)2. The selective Src family kinase inhibitors PP2 and SU6656 blocked LB- or Cyt D-mediated Ca2+ mobilisation and suppressed the upregulatory effects on AA release and 5-LO product synthesis, without affecting AA metabolism evoked by ionophore alone. We conclude that in PMNL, inhibitors of actin polymerisation cause enhancement of intracellular Ca2+ levels through Src family kinase signaling, thereby facilitating stimulus-induced release of AA and 5-LO product formation. PMID:16126002

Fischer, Lutz; Poeckel, Daniel; Buerkert, Eva; Steinhilber, Dieter; Werz, Oliver

2005-09-15

101

Influence of polyclonal immunoglobulins on the polymorphonuclear leukocyte response to lipopolysaccharide of Salmonella enteritidis as measured with luminol-enhanced chemiluminescence.  

PubMed Central

In gram-negative sepsis, the activation of polymorphonuclear leukocytes (PMN) by lipopolysaccharide (LPS) and the resulting production of superoxide and other oxygen radicals may be an important cause of tissue damage. A suppression of the PMN response to LPS stimulation would be therapeutically beneficial. The aim of this study was to determine whether different polyclonal immunoglobulins (Igs; 5S-Ig, 7S-Ig, and 19S-Ig) influence the PMN response to LPS of Salmonella enteritidis in vitro. The respiratory burst activity of PMN was measured with luminol-enhanced chemiluminescence. After addition of a 5S-Ig solution containing F(ab')2 fragments of IgG and a 19S-Ig solution containing 12% polyclonal IgM, luminol-enhanced chemiluminescence was reduced by 27% (P < 0.05) and 46% (P < 0.005), respectively. However, after addition of a 7S-Ig solution containing polyclonal IgG, luminol-enhanced chemiluminescence was increased fourfold (P < 0.05). The results suggest that the influence of polyclonal Igs on PMN response to LPS stimulation is dependent on the Ig class, F(ab')2 fragments of IgG and IgM leading to LPS neutralization and IgG leading to the production of potentially toxic oxygen radicals. PMID:7927690

Wagner, D R; Heinrich, D

1994-01-01

102

Role of the capsular polysaccharide-like serotype-specific antigen in resistance of Actinobacillus actinomycetemcomitans to phagocytosis by human polymorphonuclear leukocytes.  

PubMed Central

Serotype b-specific polysaccharide antigen (SPA) of Actinobacillus actinomycetemcomitans Y4 consists of D-fucose and L-rhamnose. To clarify the role of SPA in phagocytosis of the organism by human polymorphonuclear leukocytes (PMNs), monoclonal antibodies (MAbs) against SPA and SPA-defective mutants, which were constructed by inserting the transposon Tn916 into strain Y4, were used in a chemiluminescence (CL) assay and a phagocytic killing assay. The CL responses of human PMNs to strain Y4 were very low, and the organism was not killed by PMNs. In contrast, SPA-defective mutants induced strong CL responses. The addition of immunoglobulin G MAbs against Y4 SPA enhanced significantly both the CL responses to strain Y4 and the killing of the organism in the presence of complement. The CL responses to SPA-defective mutants were little affected by the addition of these MAbs. We conclude that SPA of A. actinomycetemcomitans plays an important role in the resistance to host defenses by PMNs. PMID:7591110

Yamaguchi, N; Kawasaki, M; Yamashita, Y; Nakashima, K; Koga, T

1995-01-01

103

Myeloperoxidase modulates the phagocytic activity of polymorphonuclear neutrophil leukocytes. Studies with cells from a myeloperoxidase-deficient patient.  

PubMed Central

Patients lacking the primary granulae enzyme, myeloperoxidase (MPO), do not usually show any increased susceptibility to infection or altered inflammatory response, in contrast to several other biochemical defects in polymorphonuclear neutrophils. We have now evaluated the role of MPO on phagocyte function in a patient with complete MPO deficiency suffering from generalized pustular psoriasis. We found that the MPO-deficient neutrophils showed enhanced phagocytosis (greater than 200% of normal) of IgG- and C3b-opsonized yeast particles and prolonged N-formylmethionyl-leucyl-phenylaline-mediated stimulation of superoxide production. When purified human MPO was added to normal neutrophils during cell adhesion, their Fc- and C3b-mediated phagocytosis was reduced without affecting cell viability. 1 microgram/ml of MPO reduced the Fc and C3b phagocytosis to 47 and 65%, respectively, whereas 10 micrograms/ml reduced the activity to 20 and 54%. Both attachment and ingestion were reduced to a similar extent, indicating that MPO affected the receptor function per se. When MPO was added to the hyperactive MPO-deficient cells, phagocytosis was reduced more rapidly. Catalase, azide, and methionine eliminated the inhibitory effect, and catalase and methionine, in fact, enhanced the phagocytic activity of adherent neutrophils. These data indicate that, apart from being a potent antimicrobial system, the oxidizing activity of the MPO-H2O2-halide system may modulate the inflammatory response by impairing certain receptor-mediated recognition mechanisms of phagocytic cells, which otherwise could elicit inflammatory reactions and tissue injury. PMID:6321554

Stendahl, O; Coble, B I; Dahlgren, C; Hed, J; Molin, L

1984-01-01

104

Effects of allogeneic leukocytes in blood transfusions during cardiac surgery on inflammatory mediators and postoperative complications  

Microsoft Academic Search

Objective: To investigate whether the higher prevalence of postoperative complications in cardiac surgery after transfusion of leukocyte-containing red blood cells can be related to inflammatory mediators. Design: Analysis of inflammatory markers interleukin-6, interleukin-10, interleukin-12, and procalcitonin in patients participating in a randomized trial comparing leukocyte-depleted with leukocyte-containing, buffy-coat-depleted red blood cells. Setting: Two university-affiliated hospitals in the Netherlands. Subjects: A

Yavuz M. Bilgin; Watering van de L. M. G; Michel I. M. Versteegh; Oers van M. H. J; Anneke Brand

2010-01-01

105

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

Microsoft Academic Search

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

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

1998-01-01

106

Antibacterial Proteins and Peptides in Nurse Shark (Ginglymostoma Cirratum) Peripheral Blood Leukocytes  

Microsoft Academic Search

In many vertebrate and invertebrate species mediators of innate immunity include antimicrobial peptides (AMPs) such as peptide fragments of histones and other proteins with previously ascribed different functions. Shark AMPs have not been described and this research examines the antibacterial activity of nurse shark (Ginglymostoma cirratum) peripheral blood leukocyte lysates. Screening of lysates prepared by homogenizing unstimulated peripheral blood leukocytes

Nichole Hinds Vaughan

2011-01-01

107

Antibacterial proteins and peptides in nurse shark ( Ginglymostoma cirratum) peripheral blood leukocytes  

Microsoft Academic Search

In many vertebrate and invertebrate species mediators of innate immunity include antimicrobial peptides (AMPs) such as peptide fragments of histones and other proteins with previously ascribed different functions. Shark AMPs have not been described and this research examines the antibacterial activity of nurse shark (Ginglymostoma cirratum) peripheral blood leukocyte lysates. Screening of lysates prepared by homogenizing unstimulated peripheral blood leukocytes

Nichole Hinds Vaughan

2011-01-01

108

Derivative of wheat germ agglutinin specifically inhibits formyl-peptide-induced polymorphonuclear leukocyte chemotaxis by blocking re-expression (or recycling) of receptors  

SciTech Connect

The mechanism of action of a derivative of wheat germ agglutinin (WGA-D) which specifically and irreversibly inhibits N-formyl-methionyl-leucyl-phenylalanine (FMLP)-induced polymorphonuclear leukocyte (PMN) chemotaxis was examined. At a concentration that completely inhibited PMN chemotaxis, WGA-D had no effect on either the uptake or release of (/sup 3/H)-FMLP by PMN. Similarly, WGA-D did not affect either the short-term binding to, or internalization by, PMN of a fluoresceinated FMLP analog. WGA-D did interfere, however, with the re-expression (or recycling) of FMLP receptors by PMN that had been preincubated with 1 ..mu..M FMLP for 10 min at 4/sup 0/C. This effect was specific for WGA-D, because it was not observed when concanavalin A was used. Scatchard plot analysis of FMLP binding to PMN after receptor re-expression demonstrated that WGA-D-treated PMN had a significant diminution in the number of high affinity receptors. WGA-D-mediated inhibition of FMLP receptor re-expression was associated with inhibition of FMLP-induced PMN chemotaxis, but had no effect on either FMLP-induced PMN superoxide anion generation or degranulation. Studies using (/sup 12/%I)-WGA-D demonstrated that PMN did not internalize WGA-D spontaneously. The data indicate that WGA-D perhaps by binding to the FMLP receptor, inhibits FMLP-induced PMN chemotaxis by blocking the re-expression (or recycling) of a population of receptors required for continuous migration.

Perez, H.D.; Elfman, F.; Lobo, E.; Sklar, L.; Chenoweth, D.; Hooper, C.

1986-03-01

109

Interaction of human monocytes, macrophages, and polymorphonuclear leukocytes with zymosan in vitro. Role of type 3 complement receptors and macrophage-derived complement.  

PubMed Central

Macrophages take up zymosan in the absence of exogenous complement via receptors for iC3b (type 3 complement receptors) acting with or without lectin-like receptors for mannosyl-fucosyl-terminated glycoconjugates. We previously provided evidence that macrophages themselves secrete complement-alternative pathway components able to opsonize zymosan locally (Ezekowitz et al., J. Exp. Med. 1984. 159:244-260). We show here that covalently bound C3 cleavage products C3b and iC3b can be eluted from zymosan particles cultivated with 36-h adherent human monocytes in the absence of serum. The ligand binding site of type 3 complement receptors is involved in macrophage-zymosan interactions as shown by inhibition studies of zymosan binding and uptake with Fab fragments of anti-C3 antibodies and monoclonal antireceptor antibodies M01 and OKM10. In contrast, antibody IB4, which binds to a receptor epitope distinct from the binding site, does not inhibit zymosan uptake. Selective modulation of macrophage receptors onto anticomplement receptor antibody and mannose-rich yeast mannan, respectively, confirms that the complement and lectin-like receptors are distinct. Human polymorphonuclear leukocytes, which express receptors for complement, but are not known to secrete complement proteins, bind and ingest only exogenously opsonized zymosan. Unopsonized zymosan is a poor trigger of respiratory burst activity in neutrophils or 7-d adherent human macrophages, but induces cell aggregation and secretion of large amounts of superoxide anion when these cells are co-cultivated in serum-free medium and challenged with zymosan. Our studies indicate that complement and/or other products synthesized by macrophages at extravascular sites could play an important role in opsonization and lysis of pathogens able to activate the alternative pathway and mediate macrophage-neutrophil collaboration in first-line host defence. Images PMID:2934410

Ezekowitz, R A; Sim, R B; MacPherson, G G; Gordon, S

1985-01-01

110

Differential expression of interleukin-8 by polymorphonuclear leukocytes of two closely related species, Ovis canadensis and Ovis aries, in response to Mannheimia haemolytica infection.  

PubMed

The pneumonic lesions and mortality caused by Mannheimia haemolytica in bighorn sheep (BHS; Ovis canadensis) are more severe than those in the related species, domestic sheep (DS; Ovis aries), under both natural and experimental conditions. Leukotoxin (Lkt) and lipopolysaccharide (LPS) are the most important virulence factors of this organism. One hallmark of pathogenesis of pneumonia is the influx of polymorphonuclear leukocytes (PMNs) into the lungs. Lkt-induced cytolysis of PMNs results in the release of cytotoxic compounds capable of damaging lung tissue. Interleukin-8 (IL-8) is a potent PMN chemoattractant. The objective of the present study was to determine if there is differential expression of IL-8 by the macrophages and PMNs of BHS and DS in response to M. haemolytica. Macrophages and PMNs of BHS and DS were stimulated with heat-killed M. haemolytica or LPS. IL-8 expression by the cells was measured by enzyme-linked immunosorbent assays and real-time reverse transcription-PCR (RT-PCR). The PMNs of BHS expressed severalfold higher levels of IL-8 than those of DS upon stimulation. Lesional lung tissue of M. haemolytica-infected BHS contained significantly higher levels of IL-8 than nonlesional tissue. The bronchoalveolar lavage (BAL) fluid of infected BHS also contained higher levels of IL-8 than that of infected DS. Depletion of IL-8 reduced migration of PMNs toward BAL fluid by approximately 50%, indicating that IL-8 is integral to PMN recruitment to the lung during M. haemolytica infection. Excessive production of IL-8, enhanced recruitment of PMNs, and PMN lysis by Lkt are likely responsible for the severity of the lung lesions in M. haemolytica-infected BHS. PMID:20515932

Herndon, Caroline N; Foreyt, William J; Srikumaran, Subramaniam

2010-08-01

111

Effects of testosterone on blood leukocytes in Plasmodium berghei -infected mice  

Microsoft Academic Search

Gonadectomized male mice aged 5 weeks were given 5 mg testosterone propionate daily for 14 days. The treatment significantly decreased the number of blood leukocytes. The number of all individual types of leukocytes except basophils in vehicle-treated gonadectomized mice was increased. Testosterone-treated mice consistently had a lower number of leukocytes after being infected withPlasmodium berghei than did vehicle-treated mice. The

A. B. Kamis; J. B. Ibrahim

1989-01-01

112

Seasonal variation of peripheral blood leukocyte telomere length in Costa Rica: a population based observational study  

PubMed Central

Objectives Peripheral blood leukocyte telomere length is increasingly being used as a biomarker of aging, but its natural variation in human populations is not well understood. Several other biomarkers show seasonal variation, as do several determinants of leukocyte telomere length. We examined whether there was monthly variation in leukocyte telomere length in Costa Rica, a country with strong seasonal differences in precipitation and infection. Methods We examined a longitudinal population based cohort of 581 Costa Rican adults age 60 and above, from which blood samples were drawn between October 2006 and July 2008. Leukocyte telomere length was assayed from these samples using the quantitative PCR method. Multivariate regression models were used to examine correlations between month of blood draw and leukocyte telomere length. Results Telomere length from peripheral blood leukocytes varied by as much as 200 base pairs depending on month of blood draw, and this difference is not likely to be due to random variation. A moderate proportion of this association is statistically accounted for by month and region specific average rainfall. We found shorter telomere length associated with greater rainfall. Conclusions There are two possible explanations of our findings. First, there could be relatively rapid month-to-month changes in leukocyte telomere length. This conclusion would have implications for understanding the natural population dynamics of telomere length. Second, there could be seasonal differences in constituent cell populations. This conclusion would suggest that future studies of leukocyte telomere length use methods to account for the potential impact of constituent cell type. PMID:24615938

Rehkopf, David H; Dow, William H; Rosero-Bixby, Luis; Lin, Jue; Epel, Elissa S; Blackburn, Elizabeth H

2014-01-01

113

Exogenous phosphatidic acid with saturated short-chain fatty acyl groups induces superoxide anion release from guinea pig peritoneal polymorphonuclear leukocytes by three different mechanisms.  

PubMed

Treatment of suspensions of guinea pig peritoneal polymorphonuclear leukocytes (PMN) with four species of phosphatidate (PA) containing short-chain fatty acids induced sustained superoxide anion (O2-) production after a lag time. The rank order of efficiency of these PAs in triggering O2- production was PA8:0 [1,2-dioctanoyl-sn-glycerol-3-phosphate (GP)] > PA10:0 (1,2-didecanoyl-GP) > PA6:0 (1,2-dicaproyl-GP) > > PA12:0 (1,2-dilauroyl-GP). The O2- release from PMN stimulated with PA10:0 or PA12:0, but not with PA6:0 or PA8:0, was lowered by the addition of 1 mM extracellular Ca2+. Studies with various inhibitors showed that the mechanism of multiphasic O2- production induced by PA8:0 depended on its concentration: 1 and 3 microM PA8:0 induced O2- production constantly after a lag time through a protein kinase-dependent mechanism that was inhibited by 100 nM staurosporine. With concentrations of PA of 10 microM or more, an additional mechanism that was independent of protein kinase became operative and predominant over the protein kinase-dependent one. This protein kinase-independent mechanism was inhibited selectively by 80 microM TMB-8. Concentrations of 30, 60 and 100 microM PA first elicited transient O2- production via another protein kinase-dependent mechanism that was more sensitive to H-7 than to staurosporine, and then sustained O2- production, mainly driven by the protein kinase-independent mechanism. Metabolism of exogenously added [14C]PA8:0 in intact PMN was examined in the presence and absence of propranolol. Results suggest that PA itself is more important rather than its degradation products such as diacylglycerol, in inducing O2- production via three different mechanisms described above. PMID:9022758

Tokumura, A; Moriyama, T; Minamino, H; Hayakawa, T; Tsukatani, H

1997-01-01

114

DNA damage in peripheral blood leukocytes in tobacco users  

PubMed Central

Aim: To Quantify the DNA single-stranded breaks in the peripheral blood leukocytes (PBLs) of tobacco-habituated individuals with clinically normal mucosa and patients with oral carcinoma. Objectives: To evaluate DNA damage levels in PBLs of tobacco-habituated individuals with clinically normal mucosa and patients with oral carcinoma and compare with a control group of healthy volunteers. To evaluate the extent of DNA damage in PBLs using Single Cell Gel Electrophoresis (SCGE) in the above groups. Materials and Methods: Patients who were attending the outpatient department were enrolled in this study. A control group of 30 healthy volunteers included in Group I were selected from various age groups who are not tobacco users in any form. Thirty patients with tobacco habituation but with clinically normal mucosa were included in Group II, while 30 tobacco-habituated patients with oral squamous carcinoma were included in Group III. A biopsy was taken from the representative area and confirmed histologically. Intravenous blood samples were collected from all the groups for evaluation of the extent of DNA damage using ethidium bromide-stained slides under fluorescent microscope. The DNA tail length was calculated by subtracting the diameter from the total length. Twenty-five randomly selected cells per slide were analyzed and mean calculated. Results: The mean DNA damage levels in patients with tobacco habits were compared with that of the control group and the results were found to be statistically significant. The mean DNA damage level in PBLs between tobacco-habituated patients with normal mucosa and oral cancer patients was found to be statistically significant. The DNA damage in cancer patients was compared with the control group and the results were found to be statistically significant. Conclusion: DNA damage evaluation in PBLs by SCGE technique is a sensitive and reliable indicator of tobacco insult. PMID:25364170

Guttikonda, Venkateswara Rao; Patil, Rekha; Kumar, GS

2014-01-01

115

Adhesion of human leukocytes to biomaterials: An in vitro study using alkanethiolate monolayers with different chemically functionalized surfaces  

Microsoft Academic Search

The adhesion of human leukocytes to self-assem- bled monolayers of well-defined surface chemistry was in- vestigated in vitro. Polymorphonuclear (PMN) and mononu- clear leukocytes were isolated from human blood by centrifugation techniques. The effect on adhesion of cell activation produced by pre-incubation of leukocytes with phytohemagglutinin (PHA) and phorbol 12-myristate 13- acetate (PMA) was also studied. Gold substrates were mod-

Judite N. Barbosa; Mario A. Barbosa; Artur P. Aguas

2003-01-01

116

In vitro Activity of Two New Oral Cephalosporins, Cefixime and Cefdinir (CI 983), on Human Peripheral Mononuclear and Polymorphonuclear Leukocyte Functions  

Microsoft Academic Search

The in vitro effects of cefixime and cefdinir (CI 983), two so-called third-generation oral cephalosporin derivatives, on human polymorphonuclear and mononuclear phagocyte functions (random migration and chemotaxis, specific and nonspecific phagocytosis, nitroblue tetrazolium reduction, superoxide production, microbicidal activity) were studied. Neither antibiotic, in the range of its attainable therapeutic concentration, exhibited any toxic effect on random migration, chemotaxis, metabolic activation

A. Fietta; C. Merlini; Gialdroni Grassi

1994-01-01

117

LeukoCatch, a quick and efficient tool for the preparation of leukocyte extracts from blood  

PubMed Central

Background Whole-protein extracts from peripheral blood leukocytes are ideal for basic and clinical research. However, lack of a simple preparation technique has limited the use of such extracts. The aim of this study is to develop a simple and easy system that can selectively obtain leukocyte extracts without hemoglobin. Methods A filter that captures the leukocytes but not RBCs was set at the bottom of a 10-mL medical syringe by sandwiching it between plastic stoppers. The capturing efficiency of leukocytes with this tool, called LeukoCatch, was examined using human macrophage cells (MONO-MAC-6). The abilities of LeukoCatch system to capture the leukocyte proteins and to remove the hemoglobin from RBCs were tested by western blot analysis using human blood samples. Results This study presents the development of LeukoCatch, a novel tool that allows the preparation of leukocyte extracts from blood samples within 3 min without centrifugation. Tissue-cultured human macrophage cells were tested to determine the optimal filter numbers and pass-through frequencies of LeukoCatch, which was then applied to 2-mL blood samples. Samples were passed 2~5 times through a LeukoCatch equipped with 5 filters, washed twice with phosphate-buffered saline for red cell removal, and leukocyte proteins were extracted with 0.5 mL of elution buffer. Western blot analysis of the purified extract indicated that more than 90% of hemoglobin was removed by the LeukoCatch and that the protein recovery rate of leukocytes was at least 4 times better than that of the conventional centrifugation method. Conclusion We conclude that LeukoCatch is useful not only for diagnosis at the bedside but also for basic research using blood samples or tissue culture cells. PMID:21849019

2011-01-01

118

Establishment of a mouse primary co-culture of endometrial epithelial cells and peripheral blood leukocytes: Effect on epithelial barrier function and leukocyte survival  

Microsoft Academic Search

This study aimed to establish an in vitro co-culture model that would allow us to study the interaction between endometrial epithelial cells and immune cells. Flow cytometry analysis and cell surface marker staining were used to identify suitable immune leukocytes from a range of sources, such as intraepithelial lymphocytes (IEL), thymocytes, splenocytes and peripheral blood leukocytes. Optimizing culture conditions such

Lok Sze Ho; Lai Ling Tsang; Yiu Wa Chung; Hsiao Chang Chan

2006-01-01

119

Chronic Iodine Toxicity in Dairy Cattle: Blood Chemistry, Leukocytes, and Milk Iodide1  

Microsoft Academic Search

Preliminary data from farm herds fed excessive dietary iodide and displaying signs of iodism indicated hyperglycemia, hypocholesterolemia, and a neutrophilic- lymphopenic shift in blood leukocytes. Subsequently blood, mitk, and urine were analyzed from 90 cows in 10 herds fed normal (average 16 mg\\/cow daily) or high (average 164 mg) iodide as ethylene- diamine dihydriodide for prophylactic purposes and from one

Donald Hillman; A. R. Curtis

1980-01-01

120

Influence of chronic caprine arthritis-encephalitis virus infection on the population of peripheral blood leukocytes.  

PubMed

The influence of caprine arthritis-encephalitis (CAE) virus infection on the population of peripheral blood leukocytes in goats was evaluated. For this purpose two groups of adult dairy female goats were formed. The experimental group consisted of 17 goats, which had been naturally infected for many years. The control group comprised 29 non-infected goats, which originated from CAE-free herd. All goats were clinically healthy. Whole blood was collected and tested in hematological analyzer and light microscope to assess the total number of leukocytes and the percentage of four leukocyte populations--neutrophils, eosinophils, monocytes and lymphocytes. Then, flow cytometry with monoclonal antibodies against several surface antigens (namely CD14, CD2, B-B2, CD4, CD8h, TCR-N6, WC1-N2 and WC1-N3) was performed to assess the proportion of lymphocyte subpopulations. Statistically significant differences (alpha < or = 0.01) were observed only in the subpopulations of T lymphocytes--percentage of all subpopulations were significantly higher in the group of seropositive goats. No statistically significant differences were revealed with respect to the total number of blood leukocytes, the average percentage of blood leukocyte populations and proportions of both T and B lymphocytes. PMID:22439329

Kaba, J; Winnicka, A; Zaleska, M; Nowicki, M; Bagnicka, E

2011-01-01

121

Microfluidic aqueous two phase system for leukocyte concentration from whole blood  

PubMed Central

Leukocytes from a whole blood sample were concentrated using a microfluidic aqueous two phase system (?ATPS). Whole blood was simultaneously exposed to polyethylene glycol (PEG) and dextran (Dex) phase streams and cells were partitioned based on their differential affinity for the streams. The laminar flow characteristic of microfluidic devices was used to create zero, one, and two stable interfaces between the polymer streams. Three different patterns of three polymer streams each were evaluated for their effectiveness in concentrating leukocytes: immiscible PEG-PEG-Dex, immiscible Dex-PEG-Dex, and miscible PEG-PBS-Dex. The most effective configuration was the Dex-PEG-Dex stream pattern which on average increased the ratio of leukocytes to erythrocytes by a factor of 9.13 over unconcentrated blood. PMID:18937070

Walker, Glenn M.

2014-01-01

122

Selection of the best features for leukocytes classification in blood smear microscopic images  

NASA Astrophysics Data System (ADS)

Automatic differential counting of leukocytes provides invaluable information to pathologist for diagnosis and treatment of many diseases. The main objective of this paper is to detect leukocytes from a blood smear microscopic image and classify them into their types: Neutrophil, Eosinophil, Basophil, Lymphocyte and Monocyte using features that pathologists consider to differentiate leukocytes. Features contain color, geometric and texture features. Colors of nucleus and cytoplasm vary among the leukocytes. Lymphocytes have single, large, round or oval and Monocytes have singular convoluted shape nucleus. Nucleus of Eosinophils is divided into 2 segments and nucleus of Neutrophils into 2 to 5 segments. Lymphocytes often have no granules, Monocytes have tiny granules, Neutrophils have fine granules and Eosinophils have large granules in cytoplasm. Six color features is extracted from both nucleus and cytoplasm, 6 geometric features only from nucleus and 6 statistical features and 7 moment invariants features only from cytoplasm of leukocytes. These features are fed to support vector machine (SVM) classifiers with one to one architecture. The results obtained by applying the proposed method on blood smear microscopic image of 10 patients including 149 white blood cells (WBCs) indicate that correct rate for all classifiers are above 93% which is in a higher level in comparison with previous literatures.

Sarrafzadeh, Omid; Rabbani, Hossein; Talebi, Ardeshir; Banaem, Hossein Usefi

2014-03-01

123

Agent-Based Modeling of Endotoxin-Induced Acute Inflammatory Response in Human Blood Leukocytes  

E-print Network

Agent-Based Modeling of Endotoxin-Induced Acute Inflammatory Response in Human Blood Leukocytes Xu of endotoxin signaling at the cellular response level. The simulation results are in accordance with our prior, Calvano SE, Lowry SF, Androulakis IP (2010) Agent-Based Modeling of Endotoxin-Induced Acute Inflammatory

Androulakis, Ioannis (Yannis)

124

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

SciTech Connect

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.

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

1990-10-01

125

Inhaled thrombolytics reduce lung microclot and leukocyte infiltration after acute blood loss.  

PubMed

We showed previously that a 30% blood loss in rats, without resuscitation, caused significant accumulation of microthrombi and leukocytes within the pulmonary circulation by 24 h. We hypothesized that the microthrombi formed spontaneously as a consequence of hemorrhage-induced stasis within the low-pressure pulmonary circuit and that the leukocytes were attracted to them. This suggested that elimination of the microthrombi, using an inhaled thrombolytic agent, could prevent the neutrophil sequestration after blood loss. To test this hypothesis, we removed 30% of the calculated blood volume from isoflurane-anesthetized, male Sprague-Dawley rats (350-500 g) over 5 min and allowed them to recover. Six hours later, we re-anesthetized the rats and nebulized tissue plasminogen activator (80 or 320 µg/kg), lactated Ringer's solution (LRS), or ipratropium bromide (i-bromide) into their lungs. We used i-bromide as a control after we discovered that nebulized LRS had thrombolytic properties. At 24 h, we removed and fixed the lungs and prepared sections for immunohistochemistry using antibodies against fibrinogen (microthrombi) and CD16 (leukocytes). Digital images of each section were obtained using a confocal microscope. Pixel counts of the images showed significantly less accumulation of microthrombi and leukocytes in lungs nebulized with tissue plasminogen activator or LRS than in non-nebulized lungs or in lungs nebulized with i-bromide (P ? 0.05). Lactated Ringer's solution becomes positively charged when nebulized (unlike i-bromide), suggesting that it eliminated microthrombi by fibrin depolymerization. We confirmed this using an in vitro assay. Our results demonstrate that lyses of microthrombi that accumulate in the lung after acute blood loss prevent subsequent leukocyte sequestration. PMID:24837203

Conhaim, Robert L; Watson, Kal E; Dovi, William F; Bates, Melissa L

2014-06-01

126

Antioxidant properties and in vitro immunomodulatory effects of peppermint (Mentha x piperita l.) Essential oils in human leukocytes  

Microsoft Academic Search

Peppermint (Mentha x piperita L.) is popular in folk medicine. Its immune effects however received so far limited attention. In the present study, peppermint oils from two cultivars (RAC 541 and Laimburg) grown in Northern Italy (Valtellina and Val Formazza) were investigated for their effects on human polymorphonuclear leukocytes (PMNs) and peripheral blood mononuclear cells (PBMCs), for their antioxidant activity

Marco Cosentino; Raffaella Bombelli; Ario Conti; Maria Laura Colombo; Andrea Azzetti; Antonello Bergamaschi; Franca Marino; Sergio Lecchini; Canton Ticino

2009-01-01

127

Can Telomere Shortening in Human Peripheral Blood Leukocytes Serve as a Disease Biomarker of Friedreich's Ataxia?  

PubMed Central

Abstract Enhanced oxidative stress and inflammation contribute to telomere erosion. Friedreich's ataxia is a neurodegenerative disorder caused by a reduction in frataxin expression that results in mitochondrial dysfunction and oxidative damage. Furthermore, frataxin deficiency induces a strong activation of inflammatory genes and neuronal death. We investigated telomere length (TL) in peripheral blood leukocytes of 37 patients with Friedreich's ataxia and 36 controls. We noted a significant telomere shortening in patients with Friedreich's ataxia compared to healthy controls (p=0.03). We also found a correlation between TL and disease duration (p=0.001). Our observations lead to the hypothesis that the TL of human peripheral blood leukocytes may serve as a biomarker of Friedreich's ataxia that could be used as an outcome measure in clinical trials. Antioxid. Redox Signal. 18, 1303–1306. PMID:23146029

Vergara, Paola; Pinelli, Michele; Filla, Alessandro; De Michele, Giuseppe; Cocozza, Sergio; Monticelli, Antonella

2013-01-01

128

Selective cyclo-oxygenase-2 inhibition with celecoxib elevates blood pressure and promotes leukocyte adherence  

Microsoft Academic Search

1 Selective inhibitors of cyclo-oxygenase-2 have been shown to be eÄective anti-inflammatory drugs with reduced gastrointestinal toxicity relative to conventional nonsteroidal anti-inflammatory drugs (NSAIDs). In the present study, we examined the possibility that selective COX-2 inhibition, by blocking prostacyclin synthesis, would increase blood pressure and cause leukocyte adherence and platelet aggregation. 2 Normal rats and rats with hypertension induced by

Marcelo N. Muscara; Nathalie Vergnolle; Fina Lovren; Christopher R. Triggle; Susan N. Elliott; Samuel Asfaha; John L. Wallace

2000-01-01

129

Gene expression profiling of porcine peripheral blood leukocytes after infection with Actinobacillus pleuropneumoniae  

Microsoft Academic Search

The gene expression profile of peripheral blood leukocytes (PBL) from extreme performing pigs after infection with Actinobacillus pleuropneumoniae was analysed using a custom complementary DNA (cDNA) microarray and quantitative reverse transcription-PCR (qRT-PCR). Four high performing animals with low disease-score (HP), three low performing animals with high disease-score (LP) and one medium performing animal with medium disease-score (MP) were selected for

Ralf J. Moser; Antonio Reverter; Sigrid A. Lehnert

2008-01-01

130

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

PubMed Central

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

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

131

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

PubMed

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

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

132

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

PubMed Central

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

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

2014-01-01

133

Local L-NAME decreases blood flow and increases leukocyte adhesion via CD18.  

PubMed

Local inhibition of nitric oxide (NO) synthesis with L-arginine analogs such as NG-nitro-L-arginine methyl ester (L-NAME) decreased red blood cell velocity (VRBC) in capillaries and increased leukocyte adhesion in postcapillary venules in rat skeletal muscle. The goal of the present study was to determine the mechanism of this response to L-NAME. Using intravital videomicroscopy, we examined blood flow in the surface microvasculature of rat extensor digitorum longus muscle. L-NAME (30 mM in the pipette) locally applied to capillaries (300 microns from feeding arteriole) reduced VRBC [control VRBC = 244 +/- 53 (SE) microns/s; delta VRBC = -52 +/- 8%] and increased leukocyte adhesion (from 0.2 +/- 0.01 to 1.3 +/- 0.3 cells/100 microns) in control animals. Systemic pretreatment with fucoidan (selectin binder), superoxide dismutase and catalase (extracellular antioxidants), dimethylthiourea (intracellular antioxidant), or ketotifen (mast cell stabilizer) did not alter this response. Pretreatment with CL26, an anti-CD18 antibody, abolished the L-NAME response. Our results suggest that L-NAME increased leukocyte-endothelial interactions via an effect on CD11/CD18 or its ligand, intercellular adhesion molecule. PMID:9575930

Mitchell, D J; Yu, J; Tyml, K

1998-04-01

134

Occurrence of nonlymphoid leukocytes that are not derived from blood islands in Xenopus laevis larvae.  

PubMed

Previous immunohistochemical observations using the monoclonal antibody (XL-1) which recognizes all types of leukocytes in Xenopus laevis revealed the occurrence of XL-1+ cells in the mesenchyme throughout the early larval body, before the appearance of any lymphocytes. The present experiments were performed to determine whether these leukocytes originate, like lymphocytes and red blood cells (RBCs), in the ventral blood islands (VBI) or the dorsolateral plate (DLP). For tracing the derivation of cells, a specific staining by quinacrine to nuclei of X. laevis and Xenopus borealis hybrid (LB) cells was used to distinguish them from X. laevis (LL) cells. Orthotopic graftings of VBI tissue from st.22-23 LB embryos to the stage-matched LL embryos and examinations at st.44-45 before differentiation of the lymphocytes showed that the proportion of XL-1+ LB cells was always significantly lower than that of RBCs with the same marker in all experimental larvae. The head (LB)-body (LL) chimeras from st.22-23 embryos and culture of the head-portions as VBI- and DLP-free explants from st.14-23 embryos both demonstrated that a significant number of XL-1+ cells which had originated in the head portions had begun to differentiate by st.42-43. These results indicate that there is a significant population of larval nonlymphoid leukocytes (mostly macrophages) that do not originate from either the VBI or DLP region, and are distributed in the mesenchyme throughout the body. PMID:2202604

Ohinata, H; Tochinai, S; Katagiri, C

1990-09-01

135

Biophysical Description of Multiple Events Contributing Blood Leukocyte Arrest on Endothelium  

PubMed Central

Blood leukocytes have a remarkable capacity to bind to and stop on specific blood vessel areas. Many studies have disclosed a key role of integrin structural changes following the interaction of rolling leukocytes with surface-bound chemoattractants. However, the functional significance of structural data and mechanisms of cell arrest are incompletely understood. Recent experiments revealed the unexpected complexity of several key steps of cell-surface interaction: (i) ligand-receptor binding requires a minimum amount of time to proceed and this is influenced by forces. (ii) Also, molecular interactions at interfaces are not fully accounted for by the interaction properties of soluble molecules. (iii) Cell arrest depends on nanoscale topography and mechanical properties of the cell membrane, and these properties are highly dynamic. Here, we summarize these results and we discuss their relevance to recent functional studies of integrin-receptor association in cells from a patient with type III leukocyte adhesion deficiency. It is concluded that an accurate understanding of all physical events listed in this review is needed to unravel the precise role of the multiple molecules and biochemical pathway involved in arrest triggering. PMID:23750158

Robert, Philippe; Touchard, Dominique; Bongrand, Pierre; Pierres, Anne

2013-01-01

136

Different role of CD73 in leukocyte trafficking via blood and lymph vessels.  

PubMed

CD73 is involved in the extracellular ATP metabolism by dephosphorylating extracellular AMP to adenosine and thus regulating permeability of the blood vessels and leukocyte traffic into the tissues. It is also present on lymphatic vessels where its distribution and function have not been characterized. We found that CD73 is expressed on a subpopulation of afferent lymph vessels but is absent on efferent lymphatics, unlike LYVE-1 and podoplanin, which are expressed on both types of lymphatics. The extracellular nucleotide metabolism on lymphatic endothelium differs from that on blood vessel endothelium as lymphatic endothelium has lower NTPDase and higher ecto-5'-nucleotidase/CD73 activity than blood vascular endothelium. In knockout mice, the lack of CD73 on lymphocytes decreases migration of lymphocytes to the draining lymph nodes more than 50% while CD73-deficient lymph vessels mediate lymphocyte trafficking as efficiently as the wild-type lymphatics. Thus, although endothelial CD73 is important for permeability and leukocyte extravasation in blood vessels, it does not have a role in these functions on lymphatics. Instead, lymphocyte CD73 is intimately involved in lymphocyte migration via afferent lymphatic vessels. PMID:21346249

Ålgars, Annika; Karikoski, Marika; Yegutkin, Gennady G; Stoitzner, Patrizia; Niemelä, Jussi; Salmi, Marko; Jalkanen, Sirpa

2011-04-21

137

Original article Potential leukocyte attractants  

E-print Network

/ml), ascorbic acid (10-1 000 pg/ml) and CaCl2 (50-560 pg/ml) had significant leukocyte attrac- tant effects) that collagenase, ascorbic acid and Ca2+ are strong candidates as attractant con- stituents of ovarian secretions. bovine ovary / follicular fluid / polymorphonuclear leukocyte / collagenase / ascorbic acid / calcium

Paris-Sud XI, Université de

138

Prenatal Arsenic Exposure and DNA Methylation in Maternal and Umbilical Cord Blood Leukocytes  

PubMed Central

Background: Arsenic is an epigenetic toxicant and could influence fetal developmental programming. Objectives: We evaluated the association between arsenic exposure and DNA methylation in maternal and umbilical cord leukocytes. Methods: Drinking-water and urine samples were collected when women were at ? 28 weeks gestation; the samples were analyzed for arsenic using inductively coupled plasma mass spectrometry. DNA methylation at CpG sites in p16 (n = 7) and p53 (n = 4), and in LINE-1 and Alu repetitive elements (3 CpG sites in each), was quantified using pyrosequencing in 113 pairs of maternal and umbilical blood samples. We used general linear models to evaluate the relationship between DNA methylation and tertiles of arsenic exposure. Results: Mean (± SD) drinking-water arsenic concentration was 14.8 ± 36.2 ?g/L (range: < 1–230 ?g/L). Methylation in LINE-1 increased by 1.36% [95% confidence interval (CI): 0.52, 2.21%] and 1.08% (95% CI: 0.07, 2.10%) in umbilical cord and maternal leukocytes, respectively, in association with the highest versus lowest tertile of total urinary arsenic per gram creatinine. Arsenic exposure was also associated with higher methylation of some of the tested CpG sites in the promoter region of p16 in umbilical cord and maternal leukocytes. No associations were observed for Alu or p53 methylation. Conclusions: Exposure to higher levels of arsenic was positively associated with DNA methylation in LINE-1 repeated elements, and to a lesser degree at CpG sites within the promoter region of the tumor suppressor gene p16. Associations were observed in both maternal and fetal leukocytes. Future research is needed to confirm these results and determine if these small increases in methylation are associated with any health effects. PMID:22466225

Baccarelli, Andrea; Hoffman, Elaine; Tarantini, Letizia; Quamruzzaman, Quazi; Rahman, Mahmuder; Mahiuddin, Golam; Mostofa, Golam; Hsueh, Yu-Mei; Wright, Robert O.; Christiani, David C.

2012-01-01

139

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

NASA Astrophysics Data System (ADS)

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 could be suggested that the visible light susceptible leukocyte reflect a pre-inflammatory state of their donors.

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

1998-12-01

140

Comparison of photonic and electromagnetic effects on the human leukocyte  

NASA Astrophysics Data System (ADS)

The dielectric and magnetic influence on human cells have been widely studied previously by the authors. Recently, the effects of energy in the visible electromagnetic spectrum have been investigated. In this subsequent study, the photonic effects on the in vitro migration of the polymorphonuclear and mononuclear leukocytes are compared with the corresponding electromagnetic field effects. Dielectric spectra of the polymorph in the 300 KHz to 400 KHz and 700 KHz to 800 KHz range have been measured. At frequencies of 350 KHz and 720 KHz an increase in the migration of the polymorphonuclear leukocyte have been observed. This stimulation was attributed to the charges on the nuclear surface. Recent preliminary data have shown a similar increased migration in the 20 MHz range. Photonic studies have indicated an enhanced migration for the polymorphonuclear leukocytes at a wavelength of 660 nm (red) and an inhibited migration at 565 nm (green). The photonic effects were postulated to be the results of a biochemical interaction rather than a membranous surface charge displacement secondary to an electric field. The migration of the white blood cells were measurement via the Boyden chamber technique and expressed in terms of a cytokinetic index which expresses the cellular movement independent of its environmental concentration gradient.

DellaVecchia, Michael A.; Beard, Richard B.; Feng, D.; Dai, Xiaoyan; Pourrezaei, Kambiz; Priezzhev, Alexander V.

1998-06-01

141

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

NASA Astrophysics Data System (ADS)

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.

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

2000-04-01

142

Effect of vitamins A and E on nitric oxide production by blood mononuclear leukocytes from neonatal calves fed milk replacer.  

PubMed

This study evaluated the effects of dietary vitamin A and E on the in vitro capacity of blood mononuclear leukocytes from calves to produce nitric oxide. Calves fed milk replacer received 100 IU/d of vitamin E as RRR-alpha-tocopherol or RRR-alpha-tocopheryl acetate and 0, 1700, 34,000, or 68,000 IU of vitamin A as retinyl acetate. Leukocytes from calves produced greater amounts of nitric oxide relative to leukocytes from adult cattle. The greater production of nitric oxide by calf leukocytes may be typical of the immature neonatal immune system. Nitric oxide production by calves fed RRR-alpha-tocopherol and either 1700 or 34,000 IU of vitamin A was less than that of calves in other groups and was more typical of production by leukocytes from cows. Our data suggest that optimal amounts of dietary vitamins A and E prompt the maturation of this response toward one that is more typical of adult cattle. Leukocytes from 1-wk-old calves produced less nitric oxide and were less responsive to stimuli than were leukocytes from older calves, a possible consequence of suppressive factors that were present in the ingested colostrum or in the circulation at birth. PMID:9891273

Rajaraman, V; Nonnecke, B J; Franklin, S T; Hammell, D C; Horst, R L

1998-12-01

143

Sildenafil prevents indomethacin-induced gastropathy in rats: role of leukocyte adherence and gastric blood flow  

PubMed Central

Nitric oxide (NO) is an important mediator of gastric mucosal defense. Sildenafil (SILD), a cyclic GMP-specific phosphodiesterase inhibitor, promotes an increase in cGMP concentrations in the gastrointestinal tract. cGMP mediates many of the biological actions of NO.We tested the hypothesis that SILD could increase mucosal defense against indomethacin-induced gastropathy in rats.SILD (1, 4 or 10?mg?kg?1, p.o.) pretreatment significantly reduced (P<0.01) the gastric damage and the increase in gastric myeloperoxidase (MPO) activity elicited by indomethacin (20?mg?kg?1 p.o.), with the maximal effect at the dose of 10?mg?kg?1.L-NAME (3, 10 or 20?mg?kg?1, i.p.) dose dependently reversed the protective effects of SILD, an effect not seen when L-arginine (L-ARG) (200?mg?kg?1, i.p.) was co-administered with L-NAME.Indomethacin-induced leukocyte adhesion, assessed by intravital microscopy, was decreased (P<0.01) by SILD, and this effect was reversed by L-NAME cotreatment.Indomethacin elicited a decrease in gastric blood flow and in gastric PGE2 levels. SILD was able to prevent the decrease in gastric blood flow (P<0.01), without diminishing the inhibitory effect of indomethacin on prostaglandin synthesis.These results indicate that SILD, acting via NO-dependent mechanisms, prevents indomethacin-induced gastropathy, possibly through a reduction of leukocyte adhesion and maintenance of gastric blood flow. PMID:16113693

Santos, Camila L; Souza, Marcellus H L P; Gomes, Antoniella S; Lemos, Henrique P; Santos, Armenio A; Cunha, Fernando Q; Wallace, John L

2005-01-01

144

Isoprinosine and levamisole as stimulators of interferon production in blood leukocytes of patients with alcoholic liver cirrhosis.  

PubMed

Blood leukocytes of 16 patients with alcoholic liver cirrhosis and 18 healthy controls were induced for interferon (IFN) production by phytohemagglutinin (PHA) and concanavalin A (ConA) in the presence or absence of isoprinosine and levamisole at concentrations of 10 micrograms/ml and 1 ng/ml. This interferon was neutralized in 87-95% by anti-HuIFN-gamma monoclonal antibodies. In the presence of the drugs the IFN-gamma production was enhanced, however, IFN-gamma titers yielded from leukocytes of cirrhotic patients were still below the titers observed in stimulated and unstimulated blood leukocytes of healthy controls. For example, IFN titers induced by PHA in the presence of levamisole (1 ng/ml) in cirrhotic patients were 2.5 times lower (20.2 +/- 11.1 U/ml) in comparison to healthy subjects (50.6 +/- 27.3 U/ml). PMID:9597085

Daniluk, J; Kandefer-Szersze?, M

1997-01-01

145

Levels of DNA damage in blood leukocyte samples from non-diabetic and diabetic female rats and their fetuses exposed to air or cigarette smoke  

Microsoft Academic Search

The objective of the present study was to evaluate DNA damage level in blood leukocytes from diabetic and non-diabetic female Wistar rats exposed to air or to cigarette smoke, and to correlate the findings with levels of DNA damage detected in blood leukocyte samples from their fetuses. A total of 20 rats were distributed into four experimental groups: non-diabetic (control;

Paula Helena Ortiz Lima; Débora Cristina Damasceno; Yuri Karen Sinzato; Maricelma da Silva Soares de Souza; Daisy Maria Fávero Salvadori; Iracema de Mattos Paranhos Calderon; Marilza Vieira Cunha Rudge

2008-01-01

146

Individual-specific variation of gene expression in peripheral blood leukocytes.  

PubMed

DNA microarray technology is used to determine gene expression profiles of various cell types, especially abnormal cells, such as cancer. By contrast, relatively little attention has been given to expression profiling of normal tissues. Here we describe studies of gene expression in peripheral blood leukocytes (PBL) from normal individuals sampled multiple times over periods ranging from several weeks up to 6 months. We demonstrate stable patterns of gene expression that differ between individuals. Among the genes whose expression varies by individual is a group of genes responsive to interferon stimulation. Certain individuals ( approximately 10-20% of those tested) showed higher baseline levels and lower inducibility of these genes in response to in vitro interferon stimulation. These studies demonstrate the feasibility of using DNA microarrays to measure the variations in gene expression of PBL from different individuals in response to environmental and genetic factors. PMID:15177552

Radich, Jerald P; Mao, Mao; Stepaniants, Sergey; Biery, Matt; Castle, John; Ward, Terry; Schimmack, Greg; Kobayashi, Sumire; Carleton, Michael; Lampe, Johanna; Linsley, Peter S

2004-06-01

147

Original article Apoptosis of polymorphonuclear leukocytes  

E-print Network

PMN was observed by histochemical stain- ing for myeloperoxidase (MPO) and electron microscopy the stimulation. Apoptosis of PMN and phagocytosis by macrophages may represent a removal mechanism, 33]. Moreover, apoptotic PMN are promptly removed by the macrophages (MAC) before they can support

Paris-Sud XI, Université de

148

RNA-seq Transcriptional Profiling of Peripheral Blood Leukocytes from Cattle Infected with Mycobacterium bovis.  

PubMed

Bovine tuberculosis, caused by infection with Mycobacterium bovis, is a major endemic disease affecting cattle populations worldwide, despite the implementation of stringent surveillance and control programs in many countries. The development of high-throughput functional genomics technologies, including gene expression microarrays and RNA-sequencing (RNA-seq), has enabled detailed analysis of the host transcriptome to M. bovis infection, particularly at the macrophage and peripheral blood level. In the present study, we have analyzed the peripheral blood leukocyte (PBL) transcriptome of eight natural M. bovis-infected and eight age- and sex-matched non-infected control Holstein-Friesian animals using RNA-seq. In addition, we compared gene expression profiles generated using RNA-seq with those previously generated using the high-density Affymetrix(®) GeneChip(®) Bovine Genome Array platform from the same PBL-extracted RNA. A total of 3,250 differentially expressed (DE) annotated genes were detected in the M. bovis-infected samples relative to the controls (adjusted P-value ?0.05), with the number of genes displaying decreased relative expression (1,671) exceeding those with increased relative expression (1,579). Ingenuity(®) Systems Pathway Analysis (IPA) of all DE genes revealed enrichment for genes with immune function. Notably, transcriptional suppression was observed among several of the top-ranking canonical pathways including Leukocyte Extravasation Signaling. Comparative platform analysis demonstrated that RNA-seq detected a larger number of annotated DE genes (3,250) relative to the microarray (1,398), of which 917 genes were common to both technologies and displayed the same direction of expression. Finally, we show that RNA-seq had an increased dynamic range compared to the microarray for estimating differential gene expression. PMID:25206354

McLoughlin, Kirsten E; Nalpas, Nicolas C; Rue-Albrecht, Kévin; Browne, John A; Magee, David A; Killick, Kate E; Park, Stephen D E; Hokamp, Karsten; Meade, Kieran G; O'Farrelly, Cliona; Gormley, Eamonn; Gordon, Stephen V; MacHugh, David E

2014-01-01

149

Leukocyte Trafficking in Experimental Autoimmune Uveitis: Breakdown of Blood?Retinal Barrier and Upregulation of Cellular Adhesion Molecules  

Microsoft Academic Search

PURPOSE. To clarify the order of events occurring in the breakdown of the blood-retinal barrier (BRB) in experimental autoimmune uveoretinitis (EAU) and in particular to study the relationships between increased vascular permeability, upregu- lation of endothelial cell adhesion molecules, and leukocyte adhesion and infiltration during EAU. METHODS. B10.RIII mice were immunized with human inter- photoreceptor retinoid binding protein (IRBP) peptide

Heping Xu; John V. Forrester; Janet Liversidge; Isabel J. Crane

2003-01-01

150

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

NASA Technical Reports Server (NTRS)

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.

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

1996-01-01

151

Gene expression patterns in blood leukocytes discriminate patients with acute infections  

PubMed Central

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

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

152

Red blood cells are a sink for interleukin 8, a leukocyte chemotaxin.  

PubMed

IL-8 (also known as neutrophil-activating peptide 1) is recognized as a potent effector of neutrophil functions. Several different cell types that contact blood, namely T lymphocytes, monocytes, and endothelial cells, secrete this polypeptide following stimulation by cytokines, or lipopolysaccharide. Here we show that when IL-8 is added to blood it rapidly partitions from the plasma fluid to the blood cells and that erythrocytes account for the vast majority of this binding. Analysis of 125I-IL-8 binding [( ala-IL-8]77 form) to human red cells indicates a single, 5 nM Kd affinity class of binding sites, present at approximately 2,000 per red cell representing approximately 15 nmol of red cell IL-8 binding sites per liter of blood. These sites are protease sensitive. Their binding of IL-8 is rapidly reversible and does not result in receptor internalization, although bound IL-8 is resistant to extraction by pH 3 buffer at 5 degrees C. 125I-IL-8 binding to red cells was not inhibited by epidermal growth factor or interleukin 1, but was inhibited by monocyte chemotactic peptide-1, which is not a neutrophil chemotaxin, but is a member of the same family of polypeptides as IL-8. FACS analysis of IL-8-mediated mobilization of Ca2+ in neutrophils indicates that the IL-8 bound to red cells is incapable of stimulating neutrophils. Thus, red cell absorption of IL-8 may function to limit stimulation of leukocytes by IL-8 released into blood. PMID:1918386

Darbonne, W C; Rice, G C; Mohler, M A; Apple, T; Hébert, C A; Valente, A J; Baker, J B

1991-10-01

153

Surface membrane expression by human blood leukocytes and platelets of decay-accelerating factor, a regulatory protein of the complement system.  

PubMed

The decay-accelerating factor (DAF), an integral membrane protein of approximately 75,000 mol wt that regulates the stability of the C3 convertases of the classical and alternative complement pathways, was initially isolated from normal erythrocyte stroma and used to prepare a polyclonal antiserum. Previously, anti-DAF antiserum has been used to immunoprecipitate DAF from surface-labeled normal erythrocytes and to document the deficiency of DAF on the surface of erythrocytes from patients with paroxysmal nocturnal hemoglobinuria, a condition in which erythrocytes express abnormal sensitivity to complement-mediated lysis. DAF has now been demonstrated by cytofluorography with anti-DAF F(ab')2 and fluoresceinated second antibody to be present on the surface of resting polymorphonuclear leukocytes (PMN), monocytes, lymphocytes, and platelets. Populations of PMN, monocytes, and platelets each exhibited a unimodal distribution of fluorescent staining, reflecting uniform cellular expression of DAF antigen, while the lymphocyte population had a skewed pattern of staining, indicating the heterogeneous expression of DAF antigen. For platelets, the shift in mean fluorescence channel observed with cytofluorographic analysis was minimal, but the presence of surface DAF on platelets was demonstrated by specific and saturable anti-DAF F(ab')2 binding. The DAF antigen, analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of dithiothreitol-reduced anti-DAF immunoprecipitates prepared from surface-labeled, isolated populations of cells, presented a single polypeptide chain of approximately 84,000 mol wt for PMN and 75,000 to 80,000 mol wt for monocytes, T and B lymphocytes, and platelets. Thus, the complement regulatory protein, DAF, is expressed on the surface of all major types of circulating blood cells from normal donors. PMID:2581636

Nicholson-Weller, A; March, J P; Rosen, C E; Spicer, D B; Austen, K F

1985-05-01

154

Tetrameric complexes of human histocompatibility leukocyte antigen (HLA)-G bind to peripheral blood myelomonocytic cells.  

PubMed

The nonclassical MHC class I molecule human histocompatibility leukocyte antigen (HLA)-G is selectively expressed on fetal trophoblast tissue at the maternal-fetal interface in pregnancy. It has long been suggested that HLA-G may inhibit maternal natural killer (NK) cells through interaction with particular NK cell receptors (KIRs). To investigate interactions of HLA-G, we constructed phycoerythrin-labeled tetrameric complexes of HLA-G refolded with a self-peptide. These HLA-G tetramers failed to bind to NK cells and cells transfected with CD94/NKG2 and killer immunoglobulin-like NK receptors. In contrast, HLA-G tetramers did bind to peripheral blood monocytes, staining a CD16(+)CD14(mid) subset with greater intensity. On transfectants, HLA-G tetramers bound to inhibitory immunoglobulin-like transcript (ILT)2 and ILT4 receptors. However, staining in the presence of antibodies reactive with ILT receptors revealed that the interaction of HLA-G tetramers with blood monocytes was largely due to binding to ILT4. These results suggest that the primary role of HLA-G may be the modulation of myelomonocytic cell behavior in pregnancy. PMID:10190906

Allan, D S; Colonna, M; Lanier, L L; Churakova, T D; Abrams, J S; Ellis, S A; McMichael, A J; Braud, V M

1999-04-01

155

Oxidative Stress by Peripheral Blood Mononuclear Cells Is Increased in Hypertensives with an Extreme-Dipper Pattern and\\/or Morning Surge in Blood Pressure  

Microsoft Academic Search

Because oxidative stress and inflammation are known to play important roles in the pathogenesis of cardiovascular events that occur most frequently in the morning, we studied the association between reactive oxygen species (ROS) formation by polymorphonuclear leukocytes (PMNs) or mononuclear cells (MNCs) and morning blood pressure (BP) rhythm. A total of 31 hypertensives in whom ambulatory BP monitoring was performed

Kensaku Maeda; Kenichi Yasunari; Takanori Watanabe; Munehiro Nakamura

2005-01-01

156

Blood leukocyte DNA hypomethylation and gastric cancer risk in a high-risk Polish population  

PubMed Central

Global hypomethylation has been shown to increase genome instability potentially leading to increased cancer risk. We determined whether global methylation in blood leukocyte DNA was associated with gastric cancer in a population-based study on 302 gastric cancer cases and 421 age- and sex-matched controls in Warsaw, Poland, between 1994 and 1996. Using PCR-pyrosequencing, we analyzed methylation levels of Alu and LINE-1, 2 CG-rich repetitive elements, to measure global methylation levels. Gastric cancer risk was highest among those with lowest level of methylation in either Alu (OR = 1.3, 95% CI = 0.9–1.9) or LINE-1 (OR = 1.4, 95% CI = 0.9–2.0) relative to those with the highest levels, although the trends were not statistically significant. For Alu, the association was stronger among those aged 70 or older (OR = 2.6, 95% CI = 1.3–5.5, p for interaction = 0.02). We did not observe meaningful differences in the associations by other risk factors and polymorphisms examined. For LINE-1, the association tended to be stronger among individuals with a family history of cancer (OR = 3.1, 95% CI = 1.4–7.0, p for interaction = 0.01), current alcohol drinkers (OR = 1.9, 95% CI = 1.0–3.6, p for interaction = 0.05), current smokers (OR = 2.3, 95% CI = 1.1–4.6, p for interaction = 0.02), those who rarely or never consumed fruit (OR = 3.1, 95% CI = 1.2–8.1, p for interaction = 0.03), CC carriers for the MTRR Ex5+123C>T polymorphism (OR = 2.3, 95% CI = 1.2–4.4, p for interaction = 0.01) and TT carriers for the MTRR Ex15+572T>C polymorphism (OR = 1.7, 95% CI = 1.0–2.8, p for interaction = 0.06). The association was not different by sex, Helicobacter pylori infection, intake of folate, vitamin B6 and total protein and the remaining polymorphisms examined. Our results indicate that interactions between blood leukocyte DNA hypomethylation and host characteristics may determine gastric cancer risk. PMID:20099281

Hou, Lifang; Wang, Hao; Sartori, Samantha; Gawron, Andrew; Lissowska, Jolanta; Bollati, Valentina; Tarantini, Letizia; Zhang, Fang Fang; Zatonski, Witold; Chow, Wong-Ho; Baccarelli, Andrea

2010-01-01

157

Leukocyte orchestration in blood and tumour tissue following interleukin-2 based immunotherapy in metastatic renal cell carcinoma.  

PubMed

With the objective of evaluating leukocyte orchestration in situ, serial blood samples and tumour tissue core needle biopsies were obtained at baseline and repeated after 1 month of therapy, among 49 consecutive single-institution patients with metastatic renal cell carcinoma (mRCC). Patients were treated with outpatient low-dose subcutaneous interleukin 2 (IL-2) and interferon alpha (IFN-alpha) alone (n = 23) or in combination with histamine dihydrochloride (n = 26). Objective responses were achieved in ten of 49 patients (20%) with an overall median survival of 14 months and an estimated 1- to 4-year survival rate of 57, 35, 24 and 22%, respectively. Toxicity was mild to moderate with no treatment-related deaths. High numbers of blood monocytes and neutrophils were significantly correlated to short survival. By contrast, high numbers of intratumoural CD3+, CD4+, CD8+ and CD57+ lymphocytes were positively correlated to objective response and/or long-term survival. Intratumoural lymphocytes showed low zeta expression, whereas blood lymphocytes showed almost normal levels of zeta expression. Neutrophils, the most frequent peripheral blood leukocyte subset, were scarce within the tumour tissue. Intratumoural eosinophils were not observed. In progressing patients, both the absolute number and the relative composition of leukocyte subsets in blood and tumour tissue remained unaffected by cytokine therapy. However, in responding patients, cytokine therapy was followed by an absolute and relative increase in T cells in blood as well as tumour tissue, an absolute and relative reduction in neutrophils in peripheral blood and a relative reduction of intratumoural macrophages. Histamine did not influence levels of intratumoural or blood leukocyte numbers, zeta-chain expression or cytotoxicity. In conclusion, the present regimen of outpatient low-dose subcutaneous IL-2 and IFN-alpha in mRCC should attract interest based on response, survival and toxicity. In responding patients, cytokine therapy was followed by substantial changes in the blood and tumour tissue leukocyte composition, correlated to response and survival. No discernable differences in immunologic parameters studied could be detected between histamine- and nonhistamine-treated patients. PMID:15088127

Donskov, Frede; Bennedsgaard, Karen Marie; Hokland, Marianne; Marcussen, Niels; Fisker, Rune; Madsen, Hans Henrik Torp; Fode, Kirsten; von der Maase, Hans

2004-08-01

158

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

PubMed Central

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

Weise, Gesa; Stoll, Guido

2012-01-01

159

Comparative quantitation of human cytomegalovirus DNA in blood leukocytes and plasma of transplant and AIDS patients.  

PubMed Central

A new method for the quantitation of human cytomegalovirus (HCMV) DNA was used to determine the levels of viral DNA in parallel in 120 blood leukocyte (leukoDNAemia) and plasma (plasmaDNAemia) samples from 8 heart or heart-lung transplant patients and 17 AIDS patients with disseminated HCMV infection. PlasmaDNAemia was consistently associated with leukoDNAemia in both groups of patients. However, at least in the transplant patients, plasmaDNAemia was not necessarily associated with clinical symptoms, appearing later and disappearing earlier than leukoDNAemia during the course of infection. Quantitative mean levels of leukoDNAemia were mostly higher than those of plasmaDNAemia in both transplant and AIDS patients. However, in the absence of antiviral treatment, plasmaDNAemia levels were significantly higher in AIDS patients than in transplant recipients, whereas leukoDNAemia levels were not significantly different between the two groups of patients. A significant correlation was found between leukoDNAemia and plasmaDNAemia in AIDS patients, as well as in transplant recipients, although to a lesser degree. However, from a diagnostic standpoint, quantitative determination of plasmaDNAemia appears to represent a much less sensitive parameter than that of leukoDNAemia (or antigenemia) for monitoring HCMV infections and antiviral treatment. Images PMID:7852562

Gerna, G; Furione, M; Baldanti, F; Sarasini, A

1994-01-01

160

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

Microsoft Academic Search

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

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

2007-01-01

161

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

Microsoft Academic Search

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

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

2010-01-01

162

Association of hypomethylation of LINE-1 repetitive element in blood leukocyte DNA with an increased risk of hepatocellular carcinoma.  

PubMed

Global DNA hypomethylation has been associated with increased risk for cancers of the colorectum, bladder, breast, head and neck, and testicular germ cells. The aim of this study was to examine whether global hypomethylation in blood leukocyte DNA is associated with the risk of hepatocellular carcinoma (HCC). A total of 315 HCC cases and 356 age-, sex- and HBsAg status-matched controls were included. Global methylation in blood leukocyte DNA was estimated by analyzing long interspersed element-1 (LINE-1) repeats using bisulfite-polymerase chain reaction (PCR) and pyrosequencing. We observed that the median methylation level in HCC cases (percentage of 5-methylcytosine (5mC)=77.7%) was significantly lower than that in controls (79.5% 5mC) (P=0.004, Wilcoxon rank-sum test). The odds ratios (ORs) of HCC for individuals in the third, second, and first (lowest) quartiles of LINE-1 methylation were 1.1 (95% confidence interval (CI) 0.7-1.8), 1.4 (95% CI 0.8-2.2), and 2.6 (95% CI 1.7-4.1) (P for trend <0.001), respectively, compared to individuals in the fourth (highest) quartile. A 1.9-fold (95% CI 1.4-2.6) increased risk of HCC was observed among individuals with LINE-1 methylation below the median compared to individuals with higher (>median) LINE-1 methylation. Our results demonstrate for the first time that individuals with global hypomethylation measured in LINE-1 repeats in blood leukocyte DNA have an increased risk for HCC. Our data provide the evidence that global hypomethylation detected in the easily obtainable DNA source of blood leukocytes may help identify individuals at risk of HCC. PMID:21960343

Di, Jian-zhong; Han, Xiao-dong; Gu, Wen-ye; Wang, Yu; Zheng, Qi; Zhang, Pin; Wu, Hui-min; Zhu, Zhong-zheng

2011-10-01

163

Leukocyte orchestration in blood and tumour tissue following interleukin-2 based immunotherapy in metastatic renal cell carcinoma  

Microsoft Academic Search

With the objective of evaluating leukocyte orchestration in situ, serial blood samples and tumour tissue core needle biopsies were obtained at baseline and repeated after 1 month of therapy, among 49 consecutive single-institution patients with metastatic renal cell carcinoma (mRCC). Patients were treated with outpatient low-dose subcutaneous interleukin 2 (IL-2) and interferon a (IFN-a) alone ( n=23) or in combination with

Frede Donskov; Karen Marie Bennedsgaard; Marianne Hokland; Niels Marcussen; Rune Fisker; Hans Henrik Torp Madsen; Kirsten Fode; Hans von der Maase

2004-01-01

164

Age-related changes following in vitro stimulation with Rhodococcus equi of peripheral blood leukocytes from neonatal foals.  

PubMed

Rhodococcus equi is an intracellular bacterium primarily known as an equine pathogen that infects young foals causing a pyogranulomatuous pneumonia. The molecular mechanisms mediating the immune response of foals to R. equi are not fully elucidated. Hence, global genomic high-throughput tools like gene expression microarrays might identify age-related gene expression signatures and molecular pathways that contribute to the immune mechanisms underlying the inherent susceptibility of foals to disease caused by R. equi. The objectives of this study were 2-fold: 1) to compare the expression profiles at specific ages of blood leukocytes from foals stimulated with virulent R. equi with those of unstimulated leukocytes; and, 2) to characterize the age-related changes in the gene expression profile associated with blood leukocytes in response to stimulation with virulent R. equi. Peripheral blood leukocytes were obtained from 6 foals within 24 hours (h) of birth (day 1) and 2, 4, and 8 weeks after birth. The samples were split, such that half were stimulated with live virulent R. equi, and the other half served as unstimulated control. RNA was extracted and the generated cDNA was labeled with fluorescent dyes for microarray hybridizations using an equine microarray. Our findings suggest that there is age-related differential expression of genes involved in host immune response and immunity. We found induction of genes critical for host immunity against pathogens (MHC class II) only at the later time-points (compared to birth). While it appears that foals up to 8-weeks of age are able to initiate a protective inflammatory response against the bacteria, relatively decreased expression of various other immune-related genes points toward inherent diminished immune responses closer to birth. These genes and pathways may contribute to disease susceptibility in foals if infected early in life, and might thus be targeted for developing preventative or therapeutic strategies. PMID:23690962

Kachroo, Priyanka; Ivanov, Ivan; Seabury, Ashley G; Liu, Mei; Chowdhary, Bhanu P; Cohen, Noah D

2013-01-01

165

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

PubMed Central

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

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

2014-01-01

166

Leukocyte, red blood cell and morphological adaptation to moderate physical training in rats undernourished in the neonatal period  

PubMed Central

Objective To analyze the impact of moderate physical exercise on the total and differential leukocyte counts and red blood cell count of 36 sixty-day-old adult male Wistar rats subjected to early malnourishment. Methods The rats were divided in nourished (N - casein 17%) and malnourished groups (M - casein 8%) and thesegroups were then subdivided in trained (T) untrained (U) creating four groups NT, NU, MT and MU. The NT and MTgroups were submitted to moderate physical exercise using a treadmill (60 min/day, 5 days/week for 8 weeks). Onthe 1st day, before the training started T0 and 24 hours after the last training day of the week (T1 until T8), a 1 mLaliquot of blood was collected from the animals' tails for analysis. The total leukocyte count was evaluated in a cellcounter with an electronic microscope. The cyanmethemoglobin technique was used to measure the hemoglobin level. The hematocrit values were determined as a percentage using the micro-hematocrit technique with a microcapillaryreader and a cell counter was used to determine the red blood cell count. The t-test was used for statistical analysis and a p-value < 0.05 was considered significant. Data are expressed as means ± standard deviation. Results There was a significant difference in the total leukocyte count between the NT (9.1 ± 0.1) and MT groups (8.0 ± 0.1) from T1 and in neutrophils between the NT (22.1 ± 0.6) and MT groups (24.6 ± 1.8) from T7 (p < 0.05). There was no statistical significance in the hemoglobin, hematocrit and red blood cell count from T1. Conclusions According to the results of this study, moderate physical exercise seems to have induced physiologic adaptation in adult rats from T1. PMID:23049442

Viana, Marcelo Tavares; Perez, Manuella Cavalcanti; Ribas, Valdenilson Ribeiro; Martins, Gilberto de Freire; de Castro, Celia Maria Machado Barbosa

2012-01-01

167

IgG4 and release of histamine from human peripheral blood leukocytes.  

PubMed

Allergen-specific IgG4 was found in the sera from many normal, nonallergic individuals. Basophil leukocytes from donors with IgG4 antibodies to an allergen, but without IgE antibodies to the same allergen, did not release histamine when challenged with this allergen. In some cases, anti-IgG4 antiserum induced a release of histamine from the leukocytes. However, these cells also release histamine after incubation with normal rabbit serum or normal sheep serum. It is concluded that the IgG4-RAST cannot be used for the detection of IgG short-term sensitizing antibodies. PMID:6173336

van Toorenenbergen, A W; Aalberse, R C

1982-01-01

168

In vitro interferon gamma regulation of CCR-3 mRNA expression in peripheral blood leukocytes from atopic asthmatics.  

PubMed

Interferon gamma (IFN-gamma) exerts major pro-inflammatory, regulatory, and anti-inflammatory actions in immune defense responses. In asthma the infiltration of eosinophils, neutrophils, and lymphocytes is a critical event. Chemokines stimulate the migration of the susceptible subset of inflammatory cells. The chemokine receptors CCR-3 are mainly expressed in eosinophils, basophils, and Th2 cells. More recently it has been demonstrated that the IFN-gamma downregulates the expression of some chemokine receptors. IgE determinations were performed using an ELISA for total IgE Peripheral blood leukocytes from patients and controls were isolated by Ficoll-Hypaque gradient. The cells were incubated in the absence or presence of 500 IU/ml of recombinant human IFN-gamma for different times. After incubation the cells were washed and lyzed for reverse transcription-polymerase chain reaction (RT-PCR) analysis. RT-PCR was performed using a Perkin-Elmer kit. The amplified bands were run in 2% agarose gels and quantified. The basal levels of CCR-3 in asthmatic patients with IgE > 150 IU/ml tend to be higher than in controls. IFN-gamma down-regulates the expression of CCR-3 in peripheral blood leukocytes from asthmatics with IgE >150 IU/ml, when compared with the basal levels of expression. In conclusions, through the modification of the expression of CCR-3 in peripheral blood leukocytes from atopic asthmatics, IFN-gamma could exert a beneficial effect in patients with asthma, regulating the migration of some inflammatory cells involved in the pathogenesis of the disease. PMID:11699731

Bello, I; Rizo, M R; Badel, C B; Blanco, E; Valenzuela, C; del Rosario, M; Quiroga, M R; Bayard, R B; Saldivar, J C; Lopez-Saura, P

2001-06-01

169

Effect of gliotoxin on human polymorphonuclear neutrophils.  

PubMed Central

OBJECTIVES: Candida albicans is known to produce gliotoxin, which has several prominent biological effects, including immunosuppression. Interference with host defenses may arise from the effects of this toxin on leukocyte structure and function. METHODS: Flow cytometric analysis revealed that polymorphonuclear leukocytes (PMN) were more sensitive to gliotoxin than were mononuclear cells. Structural and various functional aspects of PMN exposed to gliotoxin were studied. RESULTS: Gliotoxin at (1 microgram/mL) did not affect the viability but did diminish PMN chemotaxis and reduced their ability to ingest particles. Other functional aberrations included decreased nitroblue tetrazolium dye reduction, decreased superoxide production, and release of lactoferrin suggesting by degranulation. Gliotoxin also affected the ability of PMN to kill Escherichia coli. CONCLUSIONS: This study suggests a previously unrecognized potential virulence factor of C. albicans that could contribute to persistence of yeast colonization or recurrence of symptomatic infection through diminished host resistance. PMID:9812249

Shah, D T; Jackman, S; Engle, J; Larsen, B

1998-01-01

170

Comparison of metabolic, hematological, and peripheral blood leukocyte cytokine profiles of dairy cows and heifers during the periparturient period.  

PubMed

The periparturient period presents major physiological challenges for the dairy cow. It is a period that is affected by metabolic stressors, major changes in endocrine status, and altered immune function, which together result in an increased risk of disease. Immunological, hematological, and metabolic profiles from the periparturient period of heifers (primipara) were compared with those of cows (pluripara) to test the hypothesis that at the time of calving they have qualitatively different peripheral blood profiles. Blood samples were collected from 22 Holstein-Friesian animals on 3 occasions: approximately 2 wk before calving, within 24h after calving, and approximately 2 wk after calving. Quantitative PCR was used to measure the expression of a selected set of cytokines and receptors by peripheral blood leukocytes. Additional analyses included hemoglobin concentration, red cell, platelet and white cell counts (total and differentiated), and clinical diagnostic biochemical profiles. Total leukocyte counts, neutrophils, and lymphocytes were higher in heifers than cows before calving and within 24h after calving. Alkaline phosphatase was consistently higher in heifers than cows and several significant differences were observed between the 2 groups with regards to cytokine and cytokine-receptor mRNA expression. The results warrant further investigation from the perspective of identifying risk factors for metabolic and parturient disease in dairy cattle. PMID:23462170

Jonsson, N N; Fortes, M R S; Piper, E K; Vankan, D M; de Cisneros, J Prada J; Wittek, T

2013-04-01

171

Effect of glucose concentration, osmolality, and sterilization process of peritoneal dialysis fluids on cytokine production by peripheral blood mononuclear cells and polymorphonuclear cell functions in vitro.  

PubMed

We sought to investigate the effects of high glucose concentration, osmolality, and heat sterilization of peritoneal dialysis fluids on tumor necrosis factor-alpha (TNF-alpha) production by peripheral blood mononuclear cells (PBMC) and polymorphonuclear cell (PMN) functions. Blood samples were obtained from eight healthy volunteers. PBMCs and PMNs were harvested by centrifugation with Ficoll-Hypaque (Sigma, St Louis, MO). PBMC were incubated with an equal volume of test fluids and RPMI for 4 hours (pH equilibrated), followed by incubation for 20 hours in RPMI with or without endotoxin (10 ng/mL). Total TNF-alpha production was measured by radioimmunoassay. PMNs were incubated with pH-adjusted test fluids for 30 minutes. After incubation, phagocytosis was determined by the uptake of 14C-labeled Staphylococcus aureus, oxidative burst by reduction of ferricytochrome C to ferrouscytochrome C on stimulation with phorbol myristate acetate, and enzyme release by measurement of endotoxin-stimulated bactericidal/permeability increasing factor. To study the effects of increasing glucose concentration and osmolality on PBMC and PMN functions, we compared conventional 1.5% Dianeal (1.5%D), (Baxter Healthcare Corp, Deerfield, IL) 2.5% Dianeal (2.5%D), 4.25% Dianeal (4.25%D), and control (RPMI for PBMCs and Hank's balanced salt solution for PMNs). PMNs exposed to 4.25%D exhibited an inhibition of phagocytosis, phorbol myristate acetate (PMA)-stimulated oxidative burst, and bactericidal/permeability increasing factor release compared with control, 1.5%D, or 2.5%D. To study the effects of increased osmolality when controlled for glucose concentration, we compared 1.5%D with 1.5%D in which osmolality was increased to that of 4.25%D with the addition of either sodium chloride (1.5%D+NaCl) or mannitol (1.5%D+M). High osmolality induced higher TNF-alpha production by unstimulated PBMCs and decreased TNF-alpha production by endotoxin-stimulated PBMCs. PMN functions were also inhibited by high osmolality. To study the effects of increased glucose concentration when controlled for osmolality, we compared 4.25%D with 1.5%D+NaCl and 1.5%D+M. High glucose concentration induced an increase in TNF-alpha production by unstimulated PBMCs, a decrease in TNF-alpha production by endotoxin-stimulated PBMCs, and an inhibition of PMN functions. Finally, to investigate the effects of heat sterilization, we compared 4.25%D (heat sterilized) to a filter-sterilized 4.25%D (4.25%D-F). The filter-sterilized fluid induced less changes in PBMC and PMN functions compared with the heat-sterilized fluid. These data suggest that the high glucose concentration, high osmolality, and heat sterilization of peritoneal dialysis fluids adversely affect PBMC and PMN functions. These effects could predispose continuous ambulatory peritoneal dialysis patients to peritonitis, compromise host defense during infection, and jeopardize long-term survival of the peritoneal membrane. PMID:9469498

Cendoroglo, M; Sundaram, S; Jaber, B L; Pereira, B J

1998-02-01

172

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

PubMed Central

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

zilinskas, Juozas; zekonis, Jonas; zekonis, Gediminas; Sadzeviciene, Renata; Sapragoniene, Marija; Navickaite, Justina; Barzdziukaite, Ingrida

2011-01-01

173

Cell type-specific release of matrix-metallo-proteinase-9 by bacterial chemoattractant in human blood phagocytic leukocytes  

PubMed Central

Stimulation of phagocytic leukocytes with bacterial chemoattractant resulted in the release of matrix metal-loproteinases (MMPs). Little is known about the mechanisms of bacterial chemoattractant regulation of MMP in phagocytic leukocytes. We report here that the mechanisms of the bacterial chemotactic peptidefMLP-induced MMP -9 release in monocytes appeared to be different from fMLP-stimulated MMP-9 release in neutrophils. In freshly prepared peripheral blood monocytes, fMLP induces MMP-9 release, starting at 8 h after stimulation. These functions of fMLP is accompanied by an increase in TNF? expression, and mediated through the phosphorylation of ERK1/2 in monocytes. However, neutrophil preparations that responded to fMLP with MMP-9 release did not require activation of ERK1/2 and TNF? expression. These results suggest a different role of fMLP in MMP-9 expression in neutrophils and monocytes, and the signal molecules involved in mediating this effect in human blood monocytes stimulated by bacterial chemoattractant. PMID:21394287

Doerner, Astrid M; Chen, Ling-Yu; Ye, Richard D; Yong, Jiang; Huang, Shuang; Pan, Zhixing K

2011-01-01

174

[Inhibitory activity of mite IgG4, antibody on antigen-induced histamine release from human peripheral blood leukocytes].  

PubMed

Several reports have yielded conflicting results on the role of IgG4 antibody on the surfaces of target cells in immediate type allergy. This study was performed to elucidate whether IgG4 antibody inhibits IgE-mediated histamine release from target cells after antigenic stimulation, and whether it has reaginic activity. Serum was obtained from patients with nasal allergy receiving specific immunotherapy for housedust and mites. IgE and IgG4 were enriched affinity chromatographically using monoclonal antibodies to IgE and IgG4, respectively, from the pooled sera. Both fraction revealed high antibody activity to Dermatophagoides Farinae antigen. Peripheral blood leukocytes from three non-allergic donors were passively sensitized with 100 or 300 micrograms of IgG4 according to the method of Levy and Osler with a slight modification. Minimal or no histamine release was observed from leukocytes after challenge with both mite antigen and anti-IgG4 monoclonal antibodies. Furthermore, to investigate the reaginic activity of IgG4, leukocytes from patients with nasal allergy were stimulated with anti-IgG4 antibodies. The leukocytes of only three out of twenty patients released up to 10% histamine regardless of the IgG4 concentration, while the other patients' leukocytes released minimal amounts of histamine. Two of the three above-mentioned non-allergic donors were passively sensitized with 100 or 300 micrograms of IgG4 either one hour after sensitization with 100 ngs of IgE or simultaneously with the same amount of IgE. After sensitization with 100 ngs of IgE, one showed high-grade histamine release after challenge with 0.5 micrograms/ml mite antigen and the other showed middle-grade release with 0.1 micrograms/ml mite. With the presence of 300 micrograms of IgG4, histamine release was significantly inhibited in both donors regardless of the manner of sensitization.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:1380985

Yamakoshi, T

1992-07-01

175

Leukocyte Mitochondrial DNA Copy Number in Blood Is Not Associated with Major Depressive Disorder in Young Adults  

PubMed Central

Background Major depressive disorder (MDD) is the leading cause of disability worldwide, and has significant genetic predisposition. Mitochondria may have a role in MDD and so mitochondrial DNA (mtDNA) has been suggested as a possible biomarker for this disease. We aimed to test whether the mtDNA copy number of peripheral blood leukocytes is related to MDD in young adults. Methods A case-control study was conducted with 210 MDD patients and 217 healthy controls (HC). The mtDNA copy number was measured by quantitative polymerase chain reaction (qPCR) method. Depression severity was assessed by the Hamilton-17 Depression Rating Scale (HDRS-17). Results We found no significant differences in mtDNA copy number between MDD patients and HC, though the power analysis showed that our sample size has enough power to detect the difference. There were also no significant correlations between mtDNA copy number and the clinical characteristics (such as age, age of onset, episodes, Hamilton Depression Rating Scale (HDRS) score and Global Assessment of Function Scale (GAF) score) in MDD patients. Conclusion Our study suggests that leukocyte mtDNA copy number is unlikely to contribute to MDD, but it doesn’t mean that we can exclude the possibility of involvement of mitochondria in the disease. Further studies are required to elucidate whether mtDNA can be a biomarker of MDD. PMID:24809340

He, Ying; Tang, Jinsong; Li, Zongchang; Li, Hong; Liao, Yanhui; Tang, Yanqing; Tan, Liwen; Chen, Jindong; Xia, Kun; Chen, Xiaogang

2014-01-01

176

Quantitative analysis of the suppressors of cytokine signaling 1 and 3 in peripheral blood leukocytes of patients with multiple sclerosis.  

PubMed

Multiple sclerosis (MS) is an autoimmune disease characterized by a triad of inflammation, demyelination and gliosis. Because the suppressors of cytokine signaling (Socs) regulate the immune response, we quantified SOCS1 and SOCS3 transcription in peripheral blood leukocytes of patients with MS. SOCS1 transcription decreased significantly in MS patients compared with neurologically healthy persons (0.08±0.02 vs. 1.02±0.23; p=0.0001); while SOCS3 transcription increased in MS patients compared with controls (2.76±0.66 vs. 1.03±0.27; p=0.0008). Our results showed an imbalance of SOCS1 and SOCS3 transcription in MS patients, and a moderated negative correlation between them (Spearman's r=-0.57; p=0.0003). PMID:24951315

Sedeño-Monge, Virginia; Arcega-Revilla, Raúl; Rojas-Morales, Emmanuel; Santos-López, Gerardo; Perez-García, Juan Carlos; Sosa-Jurado, Francisca; Vallejo-Ruiz, Verónica; Solis-Morales, Casandra Lucrecia; Aguilar-Rosas, Salvador; Reyes-Leyva, Julio

2014-08-15

177

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

PubMed

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

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

178

Are you in or out? Leukocyte, ion, and neurotransmitter permeability across the epileptic blood-brain barrier  

PubMed Central

Summary The credo that epileptic seizures can be initiated only by “epileptic” neurons has been recently challenged. The recognition of key astrocytic-neuronal communication, and the close interaction and crosstalk between astrocytes and brain endothelial cells, has shifted attention to the blood–brain barrier (BBB) and the “neurovascular unit.” Therefore, the pursuit of mechanisms of seizure generation and epileptogenesis now includes investigations of cerebral blood flow and permeability of cerebral microvessels. For example, leukocyte adhesion molecules at the BBB have been proposed to play a role as an initiating factor for pilocarpine-induced status epilepticus, and a viral infectionmodel with a strong BBB etiology has been used to study epileptogenesis. Finally, the fact that in nonepileptic subjects seizures can be triggered by BBB disruption, together with the antiseizure effects obtained by administration of potent antiinflammatory “BBB repair” drugs, has increased the interest in neuroinflammation; both circulating leukocytes and resident microglia have been studied in this context. The dual scope of this review is the following: (1) outline the proposed role of BBB damage and immune cell activation in seizure disorders; and (2) explain how increased cerebrovascular permeability causes neuronal misfiring. The temporal sequence linking seizures to peripheral inflammation and BBB dysfunction remains to be clarified. For example, it is still debated whether seizures cause systemic inflammation or vice versa. The topographic localization of fundamental triggers of epileptic seizures also remains controversial: Are immunologic mechanisms required for seizure generation brain-specific or is systemic activation of immunity sufficient to alter neuronal excitability? Finally, the causative role of “BBB leakage” remains a largely unresolved issue. PMID:22612806

Janigro, Damir

2014-01-01

179

[DNA extraction from coagulated human blood for application in genotyping techniques for human leukocyte antigen and immunoglobulin-like receptors].  

PubMed

The objective of this study was to standardize a method for extracting high-quality DNA from samples of coagulated blood. Forty-eight samples of human coagulated blood were used for DNA extraction by means of the EZ-DNA commercial kit (Biological Industries, Beit Haemek, Israel), the Neoscience column kit (One Lambda Inc., San Diego, CA, USA) and a modified salting-out method. Only the salting-out method was able to extract high concentrations of DNA (mean, 180 ng/(1/4)microl), which were measured using the Qubit fluorescence detector (Invitrogen, USA). This method enabled amplification of HLA (human leukocyte antigen) genes using the Luminex PCR-SSO (polymerase chain reaction - sequence-specific oligonucleotide) technology, which demands good quality DNA, and amplification of KIR (killer-cell immunoglobulin-like receptor) genes using an in-house PCR-SSP (polymerase chain reaction - sequence-specific primer) technique, which demands a specific concentration of DNA (10 ng/(1/4)microl). We concluded that the modified salting-out technique was very efficient, simple and fast for DNA extraction from human coagulated blood samples, with the aim of genotyping the HLA and KIR genes. PMID:20209349

Cardozo, Daniela Maira; Guelsin, Gláucia Andréia; Clementino, Samaia Laface; Melo, Fabiano Cavalcante de; Braga, Marco Antônio; Souza, Cleonice de; Moliterno, Ricardo Alberto; Visentainer, Jeane Eliete Laguila

2009-01-01

180

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

Microsoft Academic Search

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

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

1997-01-01

181

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

Microsoft Academic Search

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

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

1996-01-01

182

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

PubMed

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

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

1997-11-01

183

Increased soluble human leukocyte antigen-G levels in peripheral blood from climbers on Mount Everest.  

PubMed

Soluble human leukocyte antigen-G (HLA-G) is involved in maternal-fetal tolerance, transplant acceptance, and tumor escape from immunosurveillance, operating by inhibiting activity of T, antigen presenting cells (APC), and natural killer (NK) cells. HLA-G gene expression is modulated in vitro after hypoxic conditions, a situation evidenced during pregnancy and tumor progression. In extreme altitude, mountaineers are in hypoxic conditions that generate physiologic adaptative responses, some of them giving rise to pathologic signs. We performed measurements of plasma soluble HLA-G in six climbers before departure of the expedition and during their ascent to and descent from summit of Mount Everest, and in 3 Sherpas at 5300-6400 m. We found that HLA-G levels are upregulated during the ascent with a unique pattern in comparison with angiogenic/lymphangiogenic factors. Our data suggest that HLA-G has to be taken into account in the mechanisms participating in adaptation to high altitudes and reinforce hypoxia as an important factor in the regulation of HLA-G expression. PMID:20732367

Bourguignon, Michel; Yaghi, Layale; Flajollet, Sébastien; Radanne-Krawice, Irène; Rouas-Freiss, Nathalie; Lugrin, Didier; Richalet, Jean-Paul; Carosella, Edgardo D; Moreau, Philippe

2010-11-01

184

Red blood cell size is inversely associated with leukocyte telomere length in a large multi-ethnic population.  

PubMed

Although mutations in the genes encoding either the protein or RNA component of telomerase have been found in patients with various blood disorders, the impact of telomere length on hematopoiesis is less well understood for subjects from the general population. Here we have measured telomere lengths of genomic DNA isolated from circulating leukocytes of 3157 subjects, ranging from 18 to 85 years of age, enrolled in a large multiethnic population based study, the Dallas Heart Study 2. Shorter telomere lengths are marginally associated with lower red blood cell counts in this cohort, but are significantly associated with larger mean red blood cell size (as measured by the MCV), increased red blood cell distribution width (RDW), higher hemoglobin levels and lower platelet counts, even after correction for age, gender and ethnicity (p-values of <0.0001, <0.0001, 0.0009 and 0.0016, respectively). In a multiple regression model we find that telomere length is a significant covariate of MCV (p = 7.6 × 10(-8)), independent of age, ethnicity, BMI, current smoking, alcohol consumption, iron or homocysteine levels. The effect of telomere length on MCV variation is comparable to the effect of smoking or alcohol consumption and is more significant in older individuals (p = 9.2 × 10(-7) for >50 years vs. p = 0.0006 for <50 years of age). To our knowledge, this is the first report of an association between telomere length and red cell size in a large urban US population and suggests a biologic mechanism for macrocytosis of aging. PMID:23226558

Kozlitina, Julia; Garcia, Christine Kim

2012-01-01

185

Red Blood Cell Size Is Inversely Associated with Leukocyte Telomere Length in a Large Multi-Ethnic Population  

PubMed Central

Although mutations in the genes encoding either the protein or RNA component of telomerase have been found in patients with various blood disorders, the impact of telomere length on hematopoiesis is less well understood for subjects from the general population. Here we have measured telomere lengths of genomic DNA isolated from circulating leukocytes of 3157 subjects, ranging from 18 to 85 years of age, enrolled in a large multiethnic population based study, the Dallas Heart Study 2. Shorter telomere lengths are marginally associated with lower red blood cell counts in this cohort, but are significantly associated with larger mean red blood cell size (as measured by the MCV), increased red blood cell distribution width (RDW), higher hemoglobin levels and lower platelet counts, even after correction for age, gender and ethnicity (p-values of <0.0001, <0.0001, 0.0009 and 0.0016, respectively). In a multiple regression model we find that telomere length is a significant covariate of MCV (p?=?7.6×10?8), independent of age, ethnicity, BMI, current smoking, alcohol consumption, iron or homocysteine levels. The effect of telomere length on MCV variation is comparable to the effect of smoking or alcohol consumption and is more significant in older individuals (p?=?9.2×10?7 for >50 years vs. p?=?0.0006 for <50 years of age). To our knowledge, this is the first report of an association between telomere length and red cell size in a large urban US population and suggests a biologic mechanism for macrocytosis of aging. PMID:23226558

Kozlitina, Julia; Garcia, Christine Kim

2012-01-01

186

Degeneration and atrophy of the thymus of lethally irradiated dogs, rescued by transfusion of cryopreserved autologous blood leukocytes  

SciTech Connect

Dogs exposed to a fatal radiation dose of 12 Gy were rescued by transfusion of autologous blood leukocytes. A severe acute and long-lasting damage to the thymus was observed. The acute damage, as observed on the tenth day, consisted of a marked reduction in the number of lymphocytes, degeneration of Hassall's bodies, and hemorrhage. Long-term effects, observed several months after irradiation, were partial to total atrophy of the thymus. Regeneration, when it occurred, was limited to a few small isolated areas in which lymphopoiesis was supported by epithelial reticular cells. In contrast, the lymph nodes of all dogs had abundant cortical lymphopoiesis. The abundant hemopoiesis present in the marrow from the tenth day after irradiation until the end of the observation period should have provided sufficient circulating precursor cells to seed the thymus and regenerate the organ to the same extent as that observed in the other blood-forming organs. The impairment of lymphopoietic regeneration in the thymus seems to be due, therefore, to damage caused by irradiation on the specific stroma of the organ, which is not able to support such activity.

Calvo, W.; Fliedner, T.M.; Herbst, E.W.; Huegl, E.B.; Boedey, B.

1987-12-01

187

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

PubMed

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

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

2014-07-01

188

Low-temperature crystal structures of tetrakis-mu-3,5-diisopropylsalicylatobis-dimethylformamidodico pper(II) and tetrakis-mu-3,5-diisopropylsalicylatobis-diethyletheratodicopp er(II) and their role in modulating polymorphonuclear leukocyte activity in overcoming seizures.  

PubMed

Two binuclear copper(II) complexes of 3,5-diisopropylsalicylic acid were characterized by single crystal X-ray diffraction methods and examined for anti-inflammatory activity using activated polymorphonuclear leukocytes and for anticonvulsant activities using electroshock and metrazol models of seizures. These complexes were crystallized from dimethylformamide (DMF) or diethylether. Tetrakis-mu-3,5-diisopropylsalicylatobis-dimethylformamidodicop per(II) [Cu(II)2(3,5-DIPS)4(DMF)2] I is in space group P 1; a = 10.393 (2), b = 11.258 (2), c = 12.734 (2) A, alpha = 96.64 (2), beta = 92.95 (2), gamma = 94.90 (2) degrees; V = 1471.7 (4) A3; Z = 1. Tetrakis-mu-3,5-diisopropylsalicylatobis-etheratodicopper(II ) [Cu(II)2(3,5-DIPS)4(ether)2] II is in space group P 1; a = 10.409 (3), b = 11.901 (4), c = 12.687 (6) A, alpha = 91.12 (5), beta = 90.84 (5), gamma = 100.90 (4) degrees; V = 1542 (1) A3; Z = 1. The structure of I was determined at 140 K from 4361 unique reflections (I > 2sigma(1)) and refined on F2 to R1 = 0.04 and wR2 = 0.09. The structure of II was determined at 180 K from 4605 unique reflections (I > 2sigma(I)) and refined on F2 to R1 = 0.05 and wR2 = 0.13. Each compound is a crystallographically centrosymmetric binuclear complex with Cu atoms bridged by four 3,5-diisopropylsalicylate ligands related by a symmetry center [Cu-Cu(i): 2.6139 (9) A in I and 2.613 (1) in II]. The four nearest O atoms around each Cu atom form a nearly rectangular planar arrangement with the square pyramidal coordination completed by the dimethylformamide (or diethylether) oxygen atom occupying an apical position, at a distance of 2.129 (2) A in I and 2.230 (3) A in II. Each Cu atom is displaced towards the DMF (or diethylether) ligand, by 0.189 A in I and 0.184 A in II, from the plane of the four O atoms. The crystal structures of I and II are essentially similar to each other, except for the DMF or diethylether accommodation. Many disorder phenomena were found in the crystal structure of I. Copper(II)2(3,5-DIPS)4(DMF)2 inhibited polymorphonuclear leukocyte (PMNL) oxidative metabolism in vitro. This effect was concentration related and significant for concentrations higher than 10 microg or 0.68 nmol/ml. Copper(II)2(3,5-DIPS)4(DMF)2 was more active than the parent ligand, 3,5-DIPS, as has been demonstrated with copper complexes of other non-steroidal anti-inflammatory drugs. The DMF and diethylether ternary complexes of Cu(II)2(3,5-DIPS)4 were found to have anticonvulsant activity in the maximal electroshock model of grand mal epilepsy in doses ranging from 26 to 258 micromol/kg of body mass following intraperitoneal, subcutaneous, or oral treatment. The DMF ternary complex was also found to be effective in the subcutaneous injection of metrazol model of petit mal epilepsy. We conclude that both ternary copper complexes are lipophilic and bioavailable, capable of facilitating the inflammatory response to brain injury and causing the subsidence of this response in bringing about remission of these disease states. PMID:11001426

Morgant, G; Dung, N H; Daran, J C; Viossat, B; Labouze, X; Roch-Arveiller, M; Greenaway, F T; Cordes, W; Sorenson, J R

2000-07-15

189

Functional groups grafted nonwoven fabrics for blood filtration-The effects of functional groups and wettability on the adhesion of leukocyte and platelet  

NASA Astrophysics Data System (ADS)

In this work, the effects of grafted functional groups and surface wettability on the adhesion of leukocyte and platelet were investigated by the method of blood filtration. The filter materials, poly(butylene terephthalate) nonwoven fabrics bearing different functional groups including hydroxyl (OH), carboxyl (COOH), sulfonic acid group (SO3H) and zwitterionic sulfobetaine group (?N((CH3)2)(CH2)3SO3?) with controllable wettability were prepared by UV radiation grafting vinyl monomers with these functional groups. Our results emphasized that both surface functional groups and surface wettability had significant effects on the adhesion of leukocyte and platelet. In the case of filter materials with the same wettability, leukocytes adhering to filter materials decreased in the order: the surface bearing OH only > the surface bearing both OH and COOH > the surface bearing sulfobetaine group > the surface bearing SO3H, while platelets adhering to filter materials decreased as the following order: the surface bearing SO3H > the surface bearing both OH and COOH > the surface bearing OH only > the surface bearing sulfobetaine group. As the wettability of filter materials increased, both leukocyte and platelet adhesion to filter materials declined, except that leukocyte adhesion to the surface bearing OH only remained unchanged.

Yang, Chao; Cao, Ye; Sun, Kang; Liu, Jiaxin; Wang, Hong

2011-01-01

190

Living T9 glioma cells expressing membrane macrophage colony-stimulating factor produce immediate tumor destruction by polymorphonuclear leukocytes and macrophages via a "paraptosis"-induced pathway that promotes systemic immunity against intracranial T9 gliomas.  

PubMed

Cloned T9-C2 glioma cells transfected with membrane macrophage colony-stimulating factor (mM-CSF) never formed subcutaneous tumors when implanted into Fischer rats, whereas control T9 cells did. The T9-C2 cells were completely killed within 1 day through a mechanism that resembled paraptosis. Vacuolization of the T9-C2 cell's mitochondria and endoplasmic reticulum started within 4 hours after implantation. By 24 hours, the dead tumor cells were swollen and terminal deoxynucleotide transferase-mediated dUTP nick-end labeling (TUNEL)-positive. Bcl2-transduced T9-C2 cells failed to form tumors in rats. Both T9 and T9-C2 cells produced cytokine-induced neutrophil chemoattractant that recruited the granulocytes into the tumor injection sites, where they interacted with the tumor cells. Freshly isolated macrophages killed the T9-C2 cells in vitro by a mechanism independent of phagocytosis. Nude athymic rats treated with antiasialo GM1 antibody formed T9-C2 tumors, whereas rats treated with a natural killer cell (NK)-specific antibody failed to form tumors. When treated with antipolymorphonuclear leukocyte (anti-PMN) and antimacrophage antibodies, 80% of nude rats formed tumors, whereas only 40% of the rats developed a tumor when a single antibody was used. This suggests that both PMNs and macrophages are involved in the killing of T9-C2 tumor cells. Immunocompetent rats that rejected the living T9-C2 cells were immune to the intracranial rechallenge with T9 cells. No vaccinating effect occurred if the T9-C2 cells were freeze-thawed, x-irradiated, or treated with mitomycin-C prior to injection. Optimal tumor immunization using mM-CSF-transduced T9 cells requires viable tumor cells. In this study optimal tumor immunization occurred when a strong inflammatory response at the injection of the tumor cells was induced. PMID:12149220

Chen, Yijun; Douglass, Thomas; Jeffes, Edward W B; Xu, Qingcheng; Williams, Christopher C; Arpajirakul, Neary; Delgado, Christina; Kleinman, Michael; Sanchez, Ramon; Dan, Qinghong; Kim, Ronald C; Wepsic, H Terry; Jadus, Martin R

2002-08-15

191

Periodic oscillation of blood leukocytes, platelets, and reticulocytes in a patient with chronic myelocytic leukemia  

Microsoft Academic Search

A patient with chronic myelocytic leukemia had a cyclic oscillation of blood neutrophils, eosinophils, basophils, monocytes, platelets, normoblasts, and reticulocytes but not of lymphocytes. The cycle interval was 53-69 days. Except for reticulocytes all other cells cycled with neutrophils. Plasma colony-stimulating factor (CSF) oscillated out of phase with neutrophils, suggesting that granulocytopoiesis is regulated through CSF by a feedback mechanism.

G. Chikkappa; G. Borner; H. Burlington; A. D. Chanana; E. P. Cronkite; S. Ohl; M. Pavelec; J. S. Robertson

1976-01-01

192

Peripheral Blood Leukocyte Production of BDNF following Mitogen Stimulation in Early Onset and Regressive Autism  

Microsoft Academic Search

Brain-derived neurotrophic factor (BDNF) is critical for neuronal differentiation and synaptic development. BDNF is also implicated in the development of psychological disorders including depression, bipolar disorder and schizophrenia. Previously, elevated BDNF levels were observed in neonatal blood samples from infants who were later diagnosed with autism when compared with children who developed normally, suggesting that BDNF may be involved in

Amanda Enstrom; Charity Onore; Angela Tarver; Irva Hertz-Picciotto; Robin Hansen; Lisa Croen

2008-01-01

193

Changes in some blood micronutrients, leukocytes and neutrophil expression of adhesion molecules in periparturient dairy cows.  

PubMed

Dairy cows are highly susceptible to infectious diseases, like mastitis, during the period around calving. Although factors contributing to increased susceptibility to infection have not been fully elucidated, impaired neutrophil recruitment to the site of infection and changes in the concentrations of some micronutrients related with the function of the immune defence has been implicated. Most of the current information is based on studies outside the Nordic countries where the conditions for dairy cows are different. Therefore, the aim of the study was to evaluate changes in blood concentrations of the vitamins A and E, the minerals calcium (Ca), phosphorous (P), and magnesium (Mg), the electrolytes potassium (K) and sodium (Na) and the trace elements selenium (Se), copper (Cu) and zinc (Zn), as well as changes in total and differential white blood cell counts (WBC) and expression of the adhesion molecules CD62L and CD18 on blood neutrophils in Swedish dairy cows during the period around calving. Blood samples were taken from 10 cows one month before expected calving, at calving and one month after calving. The results were mainly in line with reports from other countries. The concentrations of vitamins A and E, and of Zn, Ca and P decreased significantly at calving, while Se, Cu, and Na increased. Leukocytosis was detected at calving, mainly explained by neutrophilia, but also by monocytosis. The numbers of lymphocytes tended to decrease at the same time. The mean fluorescent intensity (MFI) of CD62L and CD18 molecules on blood neutrophils remained constant over time. The proportion of CD62L+ neutrophils decreased significantly at calving. The animals were fed according to, or above, their requirements. Therefore, changes in blood levels of vitamins, minerals and trace elements were mainly in response to colostrum formation, changes in dry matter intake, and ruminal metabolism around calving. Decreased levels of vitamins A and E, and of Zn at calving might have negative implications for the functions of the immune defence. The lower proportion of CD62L+ neutrophils at calving may result in less migration of blood neutrophils into the tissues, and might contribute to the increased susceptibility to infections at this time. PMID:11455894

Meglia, G E; Johannisson, A; Petersson, L; Waller, K P

2001-01-01

194

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

PubMed

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

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

2003-01-01

195

[The indices of the peripheral blood leukocytes in subjects working within a 30-kilometer area of the Chernobyl Atomic Electric Power Station and in the workers of a chemical plant].  

PubMed

Changes in leukocytes are analyzed together with the pattern of leukogrammes, morphological and cytochemical parameters characterizing leukocytes of the peripheral blood in those persons working within the zone of radiation control and in workers of chemical plants. Criteria of high informative value have been established for identification of those groups of individuals running a risk for development of hematological pathology. PMID:10476631

Klimenko, V I; Oberenko, O A

1999-06-01

196

IL-17A Expression Is Localised to Both Mononuclear and Polymorphonuclear Synovial Cell Infiltrates  

PubMed Central

Introduction This study examines the expression of IL-17A-secreting cells within the inflamed synovium and the relationship to in vivo joint hypoxia measurements. Methods IL-17A expression was quantified in synovial tissue (ST), serum and synovial fluid (SF) by immunohistochemistry and MSD-plex assays. IL-6 SF and serum levels were measured by MSD-plex assays. Dual immunofluorescence for IL-17A was quantified in ST CD15+ cells (neutrophils), Tryptase+ (mast cells) and CD4+ (T cells). Synovial tissue oxygen (tpO2) levels were measured under direct visualisation at arthroscopy. Synovial infiltration was assessed using immunohistochemistry for cell specific markers. Peripheral blood mononuclear and polymorphonuclear cells were isolated and exposed to normoxic or 3% hypoxic conditions. IL-17A and IL-6 were quantified as above in culture supernatants. Results IL-17A expression was localised to mononuclear and polymorphonuclear (PMN) cells in inflamed ST. Dual immunoflourescent staining co-localised IL-17A expression with CD15+ neutrophils Tryptase+ mast cells and CD4+T cells. % IL-17A positivity was highest on CD15+ neutrophils, followed by mast cells and then CD4+T-cells. The number of IL-17A-secreting PMN cells significantly correlated with sublining CD68 expression (r?=?0.618, p<0.01). IL-17A SF levels correlated with IL-6 SF levels (r?=?0.675, p<0.01). Patients categorized according to tp02< or >20mmHg, showed those with low tp02<20mmHg had significantly higher IL-17A+ mononuclear cells with no difference observed for PMNs. Exposure of mononuclear and polymorphonuclear cells to 3% hypoxia, significantly induced IL-6 in mononuclear cells, but had no effect on IL-17A expression in mononuclear and polymorphonuclear cells. Conclusion This study demonstrates IL-17A expression is localised to several immune cell subtypes within the inflamed synovial tissue, further supporting the concept that IL-17A is a key mediator in inflammatory arthritis. The association of hypoxia with Il-17A expression appears to be indirect, probably through hypoxia-induced pro-inflammatory pathways and leukocyte influx within the joint microenvironment. PMID:21887369

Moran, Ellen M.; Heydrich, Rene; Ng, Chin Teck; Saber, Tajvur P.; McCormick, Jennifer; Sieper, Joachim; Appel, Heiner; Fearon, Ursula; Veale, Douglas J.

2011-01-01

197

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

PubMed Central

Background Global genomic hypomethylation is a common epigenetic event in cancer that mostly results from hypomethylation of repetitive DNA elements. Case-control studies have associated blood leukocyte DNA hypomethylation with several cancers. Because samples in case-control studies are collected after disease development, whether DNA hypomethylation is causal or just associated with cancer development is still unclear. Methods In 722 elderly subjects from the Normative Aging Study cohort, we examined whether DNA methylation in repetitive elements (Alu, LINE-1) was associated with cancer incidence (30 new cases, median follow-up: 89 months), prevalence (205 baseline cases), and mortality (28 deaths, median follow-up: 85 months). DNA methylation was measured by bisulfite pyrosequencing. Results Individuals with low LINE-1 methylation (

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

2013-01-01

198

Telomere length in peripheral blood leukocytes and lung cancer risk: a large case-control study in Caucasians.  

PubMed

Telomere dysfunction is a crucial event in malignant transformation and tumorigenesis. Telomere length in peripheral blood leukocytes has been associated with lung cancer risk, but the relationship has remained controversial. In this study, we investigated whether the association might be confounded by study of different histological subtypes of lung cancer. We measured relative telomere lengths in patients in a large case-control study of lung cancer and performed stratified analyses according to the two major histologic subtypes [adenocarcinoma and squamous cell carcinoma (SCC)]. Notably, patients with adenocarcinoma had longer telomeres than controls, whereas patients with SCC had shorter telomeres compared with controls. Long telomeres were associated with increased risk of adenocarcinoma, with the highest risk associated with female sex, younger age (<60 years), and lighter smoking (<30 pack-years). In contrast, long telomeres were protective against SCC, particularly in male patients. Our results extend the concept that telomere length affects risk of lung cancer in a manner that differs with histologic subtype. PMID:24618342

Sanchez-Espiridion, Beatriz; Chen, Meng; Chang, Joe Y; Lu, Charles; Chang, David W; Roth, Jack A; Wu, Xifeng; Gu, Jian

2014-05-01

199

[Clinical significance of adhesive properties of leukocytes and blood serum in patients with multiple sclerosis].  

PubMed

Adhesive blood properties have been studied in 100 MS patients with the help of the new method developed on the basis of the leucocyte adherence inhibition (LAI) test, which was based on the calculation of the ratio of adhesive cells to non-adhesive ones. The value obtained was called the Index of Spontaneous Adhesion (ISA), while the respective indicator reflecting the effect of adhesion strengthening under the influence of autoserum, being expressed by 30% and more, was named the Effect of the Adhesion Strengthening (ES-a). Blood samples of 54 donors and 31 patients with other neurological diseases were used as controls. Statistically significant increase of ISA values of MS patients was detected as compared to the group of donors. The highest indices of ISA and ES-a were found in the primarily progressive course and at the stage of MS exacerbation in the remitting course. Correlation between levels of adhesion and clinical features as well as parameters of clinical and humoral immunity are described. A role of membrane and soluble forms of adhesion molecules in initiation and progression of the immunologic process in MS is discussed. PMID:17172233

Larina, I V; Zhirnova, I G; Komel'kova, L V; Peresedova, A V; Tsareva, M I; Gannushkina, I V; Zavalishin, A I

2006-01-01

200

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

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

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

2006-01-01

201

Leukocyte-endothelial cell adhesive receptors.  

PubMed

The first step in leukocyte localization at inflammatory foci is their adhesion to the endothelial surface. This is a complex process mediated by several adhesive molecules expressed both on the leukocyte and endothelial membrane. In the early phases of inflammation, leukocytes transiently adhere to the vessel wall in a process termed "rolling". Rolling of leukocytes is mediated by a family of adhesive molecules called selectins, expressed both on the leukocyte and endothelial surface. Other adhesive molecules and chemotactic agents act in a complementary way, with selectins stabilizing polymorphonuclear cell adhesion and mediating their transendothelial migration into the inflamed foci. In particular, leukocyte beta 2 integrins present on the leukocyte membrane, binding to two adhesive immunoglobulins (ICAM-1 and ICAM-2) on the endothelial surface. Monocytes and lymphocytes also express the integrin VLA-4 (alpha 4 beta 1) which is the ligand of the immunoglobulin VCAM-1 on endothelial cells. It is still unknown how leukocytes can migrate through the inter-endothelial junctions. An interesting possibility is that leukocyte adhesion to endothelial cells could trigger intracellular signals that in turn can mediate junction disassembly. An understanding of the molecular mechanisms at the basis of leukocyte adhesion to the vessel wall and of their infiltration into the inflamed area could help to develop specific antagonists and a more targeted therapy for inflammatory diseases. PMID:7955623

Dejana, E; Breviario, F; Caveda, L

1994-01-01

202

Effect of antiarrhythmic drugs on In-111-labeled leukocytes: chemotaxis and adherence to nylon wool  

SciTech Connect

The influence of lidocaine (L) and procainamide (P) on the chemotactic ability and adherence to nylon wool of In-111-labeled human polymorphonuclear leukocytes (PMNs) was investigated. At the normal therapeutic levels of L (0.022 mM whole blood) or P (0.03 mM whole blood) no change in PMN function was observed. However, at and above five times the aforementioned blood levels of L, significant reduction in the chemotactic ability of PMNs was noted (P <0.005). The adverse effects of In-111 radiation appeared insignificant at all L or P concentrations during the 3-hr observation period. The labeled PMNs were resistant to the toxic effects of a higher concentration of P than that of L, and the reduction in PMN chemotaxis and adherence to nylon wool was not apparent until the P concentration reached 1.5 mM.

Thakur, M.L.; Walsh, L.J.; Zaret, B.L.; Gottschalk, A.

1982-02-01

203

Effect of antiarrhythmic drugs on In-111-labeled leukocytes: chemotaxis and adherence to nylon wool  

SciTech Connect

The influence of lidocaine (L) and procainamide (P) on the chemotactic ability and adherence to nylon wool of In-111-labeled human polymorphonuclear leukocytes (PMNs) was investigated. At the normal therapeutic levels of L (0.022 mM whole blood) or P (0.03 mM whole blood) no change in PMN function was observed. However, at and above five times the aforementioned blood levels of L, significant reduction in the chemotactic ability of PMNs was noted (p less than 0.005). The adverse effects of In-111 radiation appeared insignificant at all L or P concentrations during the 3-hr observation period. The labeled PMNs were resistant to the toxic effects of a higher concentration of P than that of L, and the reduction in PMN chemotaxis and adherence to nylon wool was not apparent until the P concentration reached 1.5 mM.

Thakur, M.L.; Walsh, L.J.; Zaret, B.L.; Gottschalk, A.

1982-02-01

204

In vitro assessment of blood compatibility: residual and dynamic markers of cellular activation.  

PubMed

The blood compatibility of materials and surfaces used for medical device fabrication is a crucial factor in their function and effectiveness. Expansion of device use into more sensitive and longer term applications warrants increasingly detailed evaluations of blood compatibility that reach beyond the customary measures mandated by regulatory requirements. A panel of tests that assess both deposition on the surface and activation of circulating blood in contact with the surface has been developed. Specifically, the ability of a surface to modulate the biological response of blood is assessed by measuring: (1) dynamic thrombin generation; (2) surface-bound thrombin activity after exposure to blood; (3) activation of monocytes, polymorphonuclear leukocytes, lymphocytes, and platelets; (4) activation of complement; and (5) adherent monocytes, polymorphonuclear leukocytes, lymphocytes, and platelets on blood-contacting surfaces. The tests were used to evaluate surfaces modified with immobilized heparin (Ension's proprietary bioactive surface) and demonstrated that the modified surfaces reduced platelet activation, leukocyte activation, and complement activation in flowing human blood. Perfusion of the surfaces with human platelet-rich plasma showed that the immobilized heparin surfaces also reduce both dynamic thrombin levels in the circulating plasma and residual thrombin generated at the material surface. PMID:22210807

Johnson, Greg; Curry, Benjamin; Cahalan, Linda; Prater, Roni; Beeler, Michael; Gartner, Mark; Biggerstaff, John; Cahalan, Patrick

2013-05-01

205

Conditions for generating and assaying human leukocyte inhibitory factor (LIF) induced by purified protein derivative (PPD).  

PubMed

This report examines a variety of experimental conditions for the production of human leukocyte inhibitory factor (LIF). The data indicate that the production of LIF as measured in the indirect capillary-tube migration inhibition assay using human polymorphonuclear indicator cells accurately reflects delayed hypersensitivity to purified protein derivative (PPD) as detected by skin-test reactivity. Mononuclear cells and semipurified lymphocytes separated from whole blood by sedimentation in Ficoll-Hypaque were able to generate LIF in response to PPD. The quantity of cells responsible for LIF production were standardized and as little as 1 x 10(6) mononuclear cells were required for detectable LIF production. LIF produced by mononuclear cells required only temporary exposure to PPD, thus eliminating the necessity for a control to which antigen was added at the end of culture to antigen-free supernatants. In addition, the removal of antigen after a brief exposure helps to avoid the possible toxic effect of some antigens on the migrating polymorphonuclear leukocytes (PMNL) population. LIF production did not require extraneous serum protein (i.e., fetal bovine serum). Further, kinetic studies indicated that LIF was detected as early as 8 hr following a 2 hr-exposure to PPD and that even a 1 hr-exposure was sufficient to generate measurable detectable quantities of LIF. After 48 hr of culture, supernatants were found to contain considerable amounts of LIF, with reactivity at dilutions as high as 1:40. PMID:45790

Maurer, B A; Siwarski, D F; Fischetti, G P; McCoy, J L; Dean, J H; Herberman, R B

1978-12-01

206

Temperature-induced transcription of inflammatory mediators and the influence of Hsp70 following LPS stimulation of southern bluefin tuna peripheral blood leukocytes and kidney homogenates.  

PubMed

Temperature is known to influence inflammatory signalling in mammals, but far less understood in fish. The aim of the present study was to explore the potential effects of temperature on innate immune signalling in head kidney and leukocyte populations of the economically important southern bluefin tuna through the identification and utilization of gene expression targets in vitro. Here, we identified the mRNA sequences of five potential inflammatory mediators - TNF? (1 and 2), IL-1?, IL-8, and Cox2 - and demonstrate induction of four - TNF? (2), IL-1?, IL-8, and Cox2 - following LPS stimulation of both peripheral blood leukocytes and head kidney homogenates in vitro by real-time quantitative PCR. Comparison of transcriptional expression in cultures held at 18 and 25 °C (both within the presumed natural temperature range of this heterothermic species) showed accelerated transcription of cytokines TNF?, IL-1? and IL-8 following LPS stimulation at 25 °C in both tissue types. Peak induction reached comparable levels for each transcript at both temperatures during the 24 h test period with only limited (if any) protraction in expression resulting from cold temperature (18 °C) incubation. Partial mRNA sequences were also identified for both the constitutively expressed and heat inducible chaperone proteins Hsc70 and Hsp70, and 24 h incubation at 25 °C was sufficient to induce Hsp70 transcription in leukocyte but not in head kidney cell populations. Taken together these findings suggest that temperature exerts influence in the timing but not the degree of an innate inflammatory response in bluefin tuna and that different cell populations have differential responsiveness to heat shock in this heterothermic species. Further, LPS stimulation failed to induce Hsp70 at either incubation temperature in leukocytes; whereas 25 °C incubation caused Hsp70 up-regulation in leukocytes with or without the presence of LPS. This suggests that Hsp70 does not play a direct role in immune responsiveness for this species and that an environmental temperature of 25 °C in excess of 24 h initiates a cellular stress response in blood cells of this organism. Lastly, a strong correlation between Hsp70 and IL-8 transcriptional expression was observed following LPS/heat shock stimulation of leukocytes and five potential heat shock response elements were subsequently identified on the gene promoter region of IL-8 indicating that heat shock co-activation of this chemokine previously identified in mammals is also likely present in fish. PMID:23439399

Polinski, Mark; Bridle, Andrew; Nowak, Barbara

2013-05-01

207

Levels of DNA damage in blood leukocyte samples from non-diabetic and diabetic female rats and their fetuses exposed to air or cigarette smoke.  

PubMed

The objective of the present study was to evaluate DNA damage level in blood leukocytes from diabetic and non-diabetic female Wistar rats exposed to air or to cigarette smoke, and to correlate the findings with levels of DNA damage detected in blood leukocyte samples from their fetuses. A total of 20 rats were distributed into four experimental groups: non-diabetic (control; G1) and diabetic exposed to filtered air (G2); non-diabetic (G3) and diabetic (G4) exposed to cigarette smoke. Rats placed into whole-body exposure chambers were exposed for 30min to filtered air (control) or to tobacco smoke generated from 10 cigarettes, twice a day, for 2 months. Diabetes was induced by a pancreatic beta-cytotoxic agent, streptozotocin (40mg/kgb.w.). At day 21 of pregnancy, each rat was anesthetized and humanely killed to obtain maternal and fetal blood samples for genotoxicity analysis using the alkaline comet assay. G2, G3 and G4 dams presented higher DNA damage values in tail moment and tail length as compared to G1 group. There was a significant positive correlation between DNA damage levels in blood leukocyte samples from G2 and G3 groups (tail moment); G3 and G4 groups (tail length) and G3 group (tail intensity) and their fetuses. Thus, this study showed the association of severe diabetes and tobacco cigarette smoke exposure did not exacerbate levels of maternal and fetal DNA damages related with only diabetes or cigarette smoke exposure. Based on the results obtained and taking into account other published data, maternal diabetes requires rigid clinical control and public health and education campaigns should be increased to encourage individuals, especially pregnant women, to stop smoking. PMID:18455954

Lima, Paula Helena Ortiz; Damasceno, Débora Cristina; Sinzato, Yuri Karen; de Souza, Maricelma da Silva Soares; Salvadori, Daisy Maria Fávero; Calderon, Iracema de Mattos Paranhos; Rudge, Marilza Vieira Cunha

2008-05-31

208

Interactions between Shiga toxins and human polymorphonuclear leukocytes  

Microsoft Academic Search

Human intestinal infections by Shiga toxin (Stx)-producing Escherichia coli cause hem- orrhagic colitis and hemolytic uremic syndrome (HUS), which represents the main cause of acute renal failure in early childhood. In HUS, Stx re- leased in the gut enter the bloodstream and are targeted to renal endothelium. The mechanism of toxin delivery is still a matter of debate, although the

Maurizio Brigotti; Domenica Carnicelli; Elisa Ravanelli; Stefania Barbieri; Francesca Ricci; Andrea Bontadini; Alberto E. Tozzi; Gaia Scavia; Alfredo Caprioli; Pier Luigi Tazzari

2008-01-01

209

Chemiluminescence and superoxide anion production by leukocytes from chronic hemodialysis patients  

Microsoft Academic Search

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

Elizabeth E Ritchey; John D Wallin; Sudhir V Shah

1981-01-01

210

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

PubMed Central

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

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

211

[The influence of melatonin and epithalon on blood leukocyte count and leukocyte alkaline phosphatase in rats under different lighting conditions during ontogenesis].  

PubMed

The effect of pineal body hormone melatonin and synthetic tetrapeptide epithalon (Ala-Glu-Asp-Gly) under different light conditions on leucocytes differential count in rats were investigated. It has been established that melatonin and epithalon decrease the level of blood leucocytes and relative content of band neutrophils in 12 months rats which was higher in the constant light more than in other photoperiod. The melatonin prevents age-specific decreasing blood lymphocytes level in standard photoperiod (12 h light/12 h darkness). Contrary to melatonin, epithalon significantly reduces the number of lymphocytes and increases the number of neutrophils in some age period. The leucocytes alkaline phosphatase activity was increased during aging. Constant light in compare with other light conditions promotes early increasing of alkaline phosphatase activity (at 12 months), associated with accelerated development of pathological process in organism. The melatonin and epithalon adjacency effect on increasing of alkaline phosphatase activity under the standard as well as natural light condition demonstrate homeostatic character of geroprotectors action furthermore depend on leucocytes functional status. PMID:19432172

Uzenbaeva, L B; Vinogradova, I A; Golubeva, A G; Niuppieva, M G; Iliukha, V A

2008-01-01

212

Evaluation of leukocyte stabilisation in TransFix®-treated blood samples by flow cytometry and transmission electron microscopy  

Microsoft Academic Search

In this report, we have evaluated the effects of a TransFix®-based stabilisation technique on leukocyte scatter characteristics, immunophenotyping, membrane permeability, absolute cell counting and morphology to extend previously reported flow cytometric data focused on the lymphocyte population. We show that scatter characteristics, immunophenotyping and absolute cell counting are well preserved, particularly in the lymphocyte population. Nevertheless, a general increase in

B. Canonico; L. Zamai; S. Burattini; V. Granger; F. Mannello; P. Gobbi; C. Felici; E. Falcieri; J. T. Reilly; D. Barnett; S. Papa

2004-01-01

213

Mononuclear Leukocyte Infiltrate in Extraplacental Membranes and Preterm Delivery  

PubMed Central

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

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

2013-01-01

214

Sour cherry (Prunus cerasus) seed extract increases heme oxygenase-1 expression and decreases proinflammatory signaling in peripheral blood human leukocytes from rheumatoid arthritis patients.  

PubMed

Sour cherry seed extract (SCE) was evaluated for its capacity to inhibit lipopolysaccharide-treated human peripheral blood T cells expressing tumor necrosis factor-alpha, and the chemokine interleukin-8. Both proteins are diagnostic biomarkers for inflammatory pathologies. Peripheral blood leukocytes from 11 rheumatoid arthritis (RA) patients and 8 healthy control subjects were co-cultured for 24h in lipopolysaccharide and the extract, then evaluated by flow cytometry for T cell activation and by enzyme-linked immunoassay for lymphocyte-associated heme oxygenase-1 (HO-1) expression. There was a dose-dependent decrease in expression of the immunophenotypes: CD3+TNF-?+, and CD3+IL8+ in cultures from RA patients to a greater extent than in cells from healthy participants. These results suggest that the extract may have a modulatory roll in RA and other inflammatory disorders via the induction of HO-1, thus abating oxidative stress and strengthening regulation of pro-inflammatory signaling pathways. PMID:24631368

Mahmoud, Fadia; Haines, David; Al-Awadhi, Rana; Dashti, Ali A; Al-Awadhi, Adel; Ibrahim, Basel; Al-Zayer, Bashayer; Juhasz, Bela; Tosaki, Arpad

2014-05-01

215

Enhanced Chemotactic and Phagocytic Activities of Leukocytes in Psoriasis Vulgaris  

Microsoft Academic Search

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

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

1978-01-01

216

Kidney from uncontrolled donors after cardiac death with one hour warm ischemic time: resuscitation by extracorporal normothermic abdominal perfusion "in situ" by leukocytes-free oxygenated blood.  

PubMed

The availability of brain death donors is restricted by many factors. Use of uncontrolled donors after cardiac death could be a promising perspective, but the limiting factor in uncontrolled donation after cardiac death is the warm ischemic time. The purpose of our work was to develop an in situ kidney preservation protocol with application of the extracorporal normothermic abdominal perfusion for organ resuscitation in uncontrolled donors after cardiac death. The main attention was paid to the elimination of leukocytes as the key damaging factor from modified donor oxygenated blood circulating in the device. In 2009, we had 10 uncontrolled donors with warm ischemic time from 45 to 92 min; a normothermic extracorporal perfusion device was applied, providing preservation and restoration of kidney after ischemic damage. In 6 out of 20 kidney recipients, graft function was recovered immediately. All kidney grafts are functioning, and to the end of the third month, the average creatinine was 118.5 ± 19.9 mM. Treatment of ischemically damaged kidney by normothermic extracorporal perfusion with leukocyte depletion before procurement seems to be a challenging protocol for expanding donors' pool and demands further study. PMID:20973824

Reznik, Oleg; Skvortsov, Andrej; Loginov, Igor; Ananyev, Alexey; Bagnenko, Sergey; Moysyuk, Yan

2011-01-01

217

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

NASA Astrophysics Data System (ADS)

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.

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

2013-04-01

218

Characterization of the impairment of the uptake of apoptotic polymorphonuclear cells by monocyte subpopulations in systemic lupus erythematosus.  

PubMed

Efficient removal of apoptotic polymorphonuclear leukocytes (PMNs) is an important step in the resolution of inflammation, which protects tissues from the noxious contents of dying cells. While the impairment of apoptotic PMNs removal has been demonstrated for macrophages in systemic lupus erythematosus (SLE), recent studies show that monocytes are also capable of such phagocytosis, although their involvement in SLE is not clear. Therefore, we characterized phagocytosis of apoptotic PMNs by monocytes in 22 patients with SLE and 22 healthy controls. Using flow cytometry we demonstrate that in SLE peripheral blood monocytes show impaired phagocytosis of autologous apoptotic PMNs, while they efficiently engulf apoptotic PMNs isolated from healthy subjects. Monocytes CD14highCD16+ and CD14dimCD16+ more efficiently interacted with apoptotic neutrophils than CD16- cells both in SLE and healthy subjects. Monocytes in SLE showed modestly decreased expression of CD35 and CD91 and increased expression of T Cell Ig- and mucin-domain-containing molecule-3 (TIM-3); however, these differences were evident mainly in selected subsets of monocytes (CD16+) while defects in phagocytosis were observed in all monocyte subsets. Apoptotic cell-dependent induction of lipopolysaccharide (LPS) stimulated production of anti-inflammatory cytokine IL-10 by peripheral blood mononuclear cells (PBMC) was blunted in SLE while the production of pro-inflammatory cytokine TNF-? was unchanged. PMID:24969081

Miko?ajczyk, T P; Skiba, D; Batko, B; Krezelok, M; Wilk, G; Osmenda, G; Pryjma, J R; Guzik, T J

2014-11-01

219

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

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

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

2014-06-01

220

Generation of large numbers of human dendritic cells from whole blood passaged through leukocyte removal filters: an alternative to standard buffy coats.  

PubMed

Many blood banks now use whole blood inline filtration to produce leukocyte-depleted blood products. As a result, a common source of large numbers of human dendritic cells (DC) for research purposes, namely standard buffy coats, has been lost. Therefore, we have adapted our conventional method for growing DC from CD14(+) precursors in order to make use of these filter units. A dextran solution containing human serum albumin was used to flush back the filters. After pelleting, mononuclear cells were obtained by standard density gradient centrifugation (Lymphoprep). To eliminate T cells, we used rosetting with sheep red blood cells. In addition to the classical PBMC, the cell population obtained after Lymphoprep centrifugation was found to contain high numbers of CD14(+) granulocytes which could be depleted by separation on an additional Percoll gradient. At this stage, FACS analysis revealed a cell population that resembled the CD14(+) monocyte-enriched population, obtained from traditional buffy coat preparations after Lymphoprep centrifugation and T cell elimination. Culture of the cells and the induction of maturation was identical to the previously described procedures, except that the culture time was reduced from 7 to 5 days and the maturation time from 3 to 2 days. Analyses of the major molecules indicative of DC maturation (CD83, CD86, CD208/DC-LAMP) and functional analyses of the T cell-stimulatory capacity of the DC population (using the MLR assay with normal peripheral T cells and naive T cells) revealed no major differences from buffy coat-derived DC preparations. PMID:11334969

Ebner, S; Neyer, S; Hofer, S; Nussbaumer, W; Romani, N; Heufler, C

2001-06-01

221

Reduced Arylsulfatase B activity in leukocytes from cystic fibrosis patients.  

PubMed

The enzyme Arylsulfatase B (ARSB; N-acetylgalactosamine-4-sulfatase) removes 4-sulfate groups from chondroitin-4-sulfate and dermatan sulfate and is required for the degradation of these sulfated glycosaminoglycans (sGAGs). Since these GAGs accumulate in patients with Cystic Fibrosis (CF), we investigated the activity of ARSB in leukocytes of patients with CF, to consider if reduced activity of ARSB might contribute to the pathophysiology of CF. Previous cell-based experiments had demonstrated that when the deficiency of the cystic fibrosis transmembrane regulator (CFTR) was corrected in bronchial epithelial cells, the ARSB activity increased significantly. De-identified, citrated blood samples were collected from 16 children with CF and 31 control subjects, seen in the Pediatric Clinic at Rush University Medical Center. Polymorphonuclear leukocytes (PMN) and mononuclear cell (MC) populations were separated by density gradient, and blinded determinations of ARSB activity were performed using the exogenous substrate 4-methylumbilliferyl sulfate. Interleukin-6 was measured in the plasma samples by ELISA. ARSB activity was significantly less in the PMN and MC from the CF patients than controls (P?

Sharma, Girish; Burke, Jenifer; Bhattacharyya, Sumit; Sharma, Neha; Katyal, Shivani; Park, R Lucy; Tobacman, Joanne

2013-03-01

222

Technetium-99m-labeled white blood cells: a new method to define the local and systemic role of leukocytes in acute experimental pancreatitis.  

PubMed Central

OBJECTIVE: We developed a new method to quantitate leukocyte accumulation in tissues and used it to examine the time course and severity of acute experimental pancreatitis. BACKGROUND: Leukocyte activation and infiltration are believed to be critical steps in the progression from mild to severe pancreatitis and responsible for many of its systemic complications. METHODS: Pancreatitis of graded severity was induced in Sprague-Dawley rats with a combination of caerulein and controlled intraductal infusion. Technetium-99m (99mTc)-labeled leukocytes were quantified in pancreas, lung, liver, spleen, and kidney and compared with myeloperoxidase activity. The severity of pancreatitis was ascertained by wet/dry weight ratio, plasma amylase, and trypsinogen activation peptide in the pancreas. The time course of leukocyte accumulation was determined over 24 hours. RESULTS: Pancreatic leukocyte infiltration correlated well with tissue myeloperoxidase concentrations. In mild pancreatitis, leukocytes accumulated only in the pancreas. Moderate and severe pancreatitis were characterized by much greater leukocyte infiltration in the pancreas than in mild disease (p < 0.01), and increased 99mTc radioactivity was detectable in the lung as early as 3 hours. 99mTc radioactivity correlated directly with the three levels of pancreatitis. CONCLUSIONS: Mild pancreatitis is characterized by low-level leukocyte activation and accumulation in the pancreas without recruitment of other organs; marked leukocyte accumulation was found in the pancreas and in the lung in more severe grades of pancreatitis. These findings provide a basis for the pathophysiologic production of cytokines and oxygen free radicals, which potentiate organ injury in severe pancreatitis. This study validates a new tool to study local and systemic effects of leukocytes in pancreatitis as well as new therapeutic hypotheses. PMID:9445115

Werner, J; Dragotakes, S C; Fernandez-del Castillo, C; Rivera, J A; Ou, J; Rattner, D W; Fischman, A J; Warshaw, A L

1998-01-01

223

The effects of stress on the enzymes of peripheral leukocytes  

NASA Technical Reports Server (NTRS)

Previous work showed an early response of rabbit and human leukocyte enzymes to the stress of bacterial infection. Since these represented a mixed population of leukocytes and since polymorphonuclear leukocytes (PMN) increased in these preparations, it was necessary to establish whether the observed increase in lactate dehydrenase (LDH) and protein was the result of an increase in any one particular cell type or in all cells. The need for the development of a simple reproducible method for the differential separation of peripheral leukocytes for the furtherance of our own studies was apparent. It was also becoming increasingly apparent that morphologically similar cells, such as small lymphocytes (L) and macrophages, were capable of different biological functions. A dextran gradient centrifugation method was developed which has provided an easily reproducible technique for separating L from PMN. During the course of this work, in which over 250 rabbits were examined, the pattern of daily leukocyte protein and enzyme variation became increasingly more apparent. This information could have some impact on future work with leukocyte enzymes, by our group and by other workers. The differences in normal protein and enzyme levels maintained by some individuals, and some inbred strains, were evaluated and reported separately. It has been shown that one type of leukocyte may react more to a given stress than other leukocytes.

Leise, E. M.; Gray, I.

1973-01-01

224

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

PubMed Central

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

2014-01-01

225

Influence of antibiotics on formylmethionyl-leucyl-phenylalanine-induced leukocyte chemiluminescence.  

PubMed Central

The effect of three antimicrobial agents, penicillin G, ampicillin, and chloramphenicol, on luminol-enhanced chemiluminescence of polymorphonuclear leukocytes stimulated by the chemoattractant formylmethionyl-leucyl-phenylalanine was studied. An inhibitory effect of penicillin G and of ampicillin was demonstrated, whereas chloramphenicol gave rise to an enhancement of the chemiluminescence response from polymorphonuclear leukocytes. These effects could be due to interaction between the drugs and the polymorphonuclear leukocytes, but they could also be the result of interference with the generation of light without any effect on the cells. Therefore, the effects of the same antimicrobial agents on the chemiluminescence generated from a cell-free system consisting of myeloperoxidase and hydrogen peroxide were investigated in parallel. The results obtained in the cell-free system were almost identical to those obtained in the cell system; i.e., penicillin G and ampicillin caused an inhibition and chloramphenicol caused an enhancement of the light emission. These results indicate that observed effects induced by drugs in a chemiluminescence assay are not necessarily due to interaction between the drug and polymorphonuclear leukocytes but may be caused by interference with other components of the assay. In view of these findings, the conflicting data reported in the literature on the effects of antimicrobial agents on phagocyte function are discussed. PMID:3606075

Briheim, G; Dahlgren, C

1987-01-01

227

Cytotoxic Activity of Peripheral Blood Mononuclear Leukocytes, Activated by Interleukin-2/?-Cyclodextrin Nanocomposition against Androgen Receptor-Negative Prostate Cancers.  

PubMed

Nanocomposition comprised of interleukin-2 in suboptimal noneffective concentration and ?-cyclodextrin was studied in vitro. This preparation as well as interleukin-2 in optimal concentration was shown to increase natural killer activity to K-562 cells and cytotoxicity of activated peripheral blood mononuclear cells (PBMCs) against PC-3 and DU 145 cells. At the same time ?-cyclodextrin or interleukin-2 in equimolar concentrations did not influence the spontaneous killer activity of PBMC. This combination of cyclodextrin + interleukin-2 led to the decrease of interleukin-2 effective concentration by an order. This phenomenon could be explained by cyclodextrins ability to promote the formation of nanoparticles with drugs, which results in enhancing their water solubility and bioavailability. Besides, interleukine-2/?-cyclodextrin nanocomposition as opposed to interleukin-2 alone led to increasing the number of not only lymphocytes, but also macrophages contained in activated PBMC population. Application of low concentration of interleukin-2 allowing for good clinical efficiency may significantly mitigate the side effects of the drug and enable to develop adoption of immunotherapy for patients with androgen-resistant prostate cancer. PMID:22084730

Anisimova, Natalia Yu; Sosnov, Andrey V; Ustyuzhanina, Nadezhda E; Baronzio, Gianfranco; Kiselevsky, Mikhail V

2011-01-01

228

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

PubMed Central

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

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

229

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

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

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

2007-05-01

230

Intracellular concentrations of micafungin in different cellular compartments of the peripheral blood.  

PubMed

Whilst micafungin serum concentrations are well studied, little is known about its concentrations within cellular compartments of the peripheral blood. Hence, in this study blood samples were collected from patients receiving micafungin (n=26). These samples were separated by double-discontinuous Ficoll-Hypaque density gradient centrifugation. Intracellular concentrations within the obtained cells, i.e. peripheral blood mononuclear cells (PBMCs), polymorphonuclear leukocytes (PMNs) and red blood cells (RBCs), were determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Within PBMCs and PMNs, the intracellular micafungin concentration was significantly increased compared with the concentration in plasma (P<0.001). The intracellular concentration within RBCs did not significantly differ from the plasma concentration. Micafungin reaches high concentrations in human PBMCs and PMNs and is present in RBCs. In vitro data showed that intracellular uptake of micafungin by PBMCs depends on the albumin concentration of the surrounding medium, but only at non-physiological protein concentrations. PMID:22230332

Farowski, Fedja; Cornely, Oliver A; Vehreschild, Jörg J; Bauer, Tim; Hartmann, Pia; Steinbach, Angela; Vehreschild, Maria J G T; Scheid, Christof; Müller, Carsten

2012-03-01

231

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

PubMed Central

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

1996-01-01

232

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

Microsoft Academic Search

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

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

1994-01-01

233

Substitution of aspartate for glycine 1018 in the type III procollagen (COL3A1) gene causes type IV Ehlers-Danlos syndrome: the mutated allele is present in most blood leukocytes of the asymptomatic and mosaic mother.  

PubMed Central

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 acid 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. Images Figure 5 Figure 2 Figure 3 Figure 4 PMID:1496983

Kontusaari, S; Tromp, G; Kuivaniemi, H; Stolle, C; Pope, F M; Prockop, D J

1992-01-01

234

Secretory virulence factors produced by Staphylococcus aureus isolates obtained from mastitic bovine milk--effect on bovine polymorphonuclear neutrophils.  

PubMed

The aim of the research was to test whether exogenic virulence factors secreted by Staphylococcus aureus isolates are involved in mechanisms that allow the bacteria to modulate and evade phagocytosis by bovine polymorphonuclear neutrophils. The research was based on the comparison of the effects of supernatants, prepared from cultures of 30 S. aureus isolates, on the functional properties of bovine neutrophils in vitro. S. aureus isolates were collected from milk samples from cows with clinical mastitis. Supernatants, which were used to treat leukocytes, were prepared from 18 h S. aureus cultures. Exogenic virulence factors secreted by S. aureus isolates significantly influenced the phagocytosis parameters evaluated. Depending on their leukotoxic or superantigenic properties, supernatants could affect the ingestion process, and also showed an influence on the digestion efficiency and phagocytosis carried out by bovine polymorphonuclear neutrophils in vitro. PMID:21944833

Fijalkowski, Karol; Czernomysy-Furowicz, Danuta; Irwin, Jane A; Nawrotek, Pawel; Pobucewicz, Anna

2012-08-01

235

Leukocyte nucleus segmentation and nucleus lobe counting  

PubMed Central

Background Leukocytes play an important role in the human immune system. The family of leukocytes is comprised of lymphocytes, monocytes, eosinophils, basophils, and neutrophils. Any infection or acute stress may increase or decrease the number of leukocytes. An increased percentage of neutrophils may be caused by an acute infection, while an increased percentage of lymphocytes can be caused by a chronic bacterial infection. It is important to realize an abnormal variation in the leukocytes. The five types of leukocytes can be distinguished by their cytoplasmic granules, staining properties of the granules, size of cell, the proportion of the nuclear to the cytoplasmic material, and the type of nucleolar lobes. The number of lobes increased when leukemia, chronic nephritis, liver disease, cancer, sepsis, and vitamin B12 or folate deficiency occurred. Clinical neutrophil hypersegmentation has been widely used as an indicator of B12 or folate deficiency.Biomedical technologists can currently recognize abnormal leukocytes using human eyes. However, the quality and efficiency of diagnosis may be compromised due to the limitations of the biomedical technologists' eyesight, strength, and medical knowledge. Therefore, the development of an automatic leukocyte recognition system is feasible and necessary. It is essential to extract the leukocyte region from a blood smear image in order to develop an automatic leukocyte recognition system. The number of lobes increased when leukemia, chronic nephritis, liver disease, cancer, sepsis, and vitamin B12 or folate deficiency occurred. Clinical neutrophil hypersegmentation has been widely used as an indicator of B12 or folate deficiency. Results The purpose of this paper is to contribute an automatic leukocyte nuclei image segmentation method for such recognition technology. The other goal of this paper is to develop the method of counting the number of lobes in a cell nucleus. The experimental results demonstrated impressive segmentation accuracy. Conclusions Insensitive to the variance of images, the LNS (Leukocyte Nuclei Segmentation) method functioned well to isolate the leukocyte nuclei from a blood smear image with much better UR (Under Segmentation Rate), ER (Overall Error Rate), and RDE (Relative Distance Error). The presented LC (Lobe Counting) method is capable of splitting leukocyte nuclei into lobes. The experimental results illuminated that both methods can give expressive performances. In addition, three advanced image processing techniques were proposed as weighted Sobel operator, GDW (Gradient Direction Weight), and GBPD (Genetic-based Parameter Detector). PMID:21073711

2010-01-01

236

Reduced Arylsulfatase B Activity in Leukocytes from Cystic Fibrosis Patients  

PubMed Central

Summary The enzyme Arylsulfatase B (ARSB; N-acetylgalactosamine-4-sulfatase) removes 4-sulfate groups from chondroitin-4-sulfate and dermatan sulfate and is required for the degradation of these sulfated glycosaminoglycans (GAGs). Since these GAGs accumulate in patients with Cystic Fibrosis (CF), we investigated the activity of ARSB in leukocytes of patients with CF, to consider if reduced activity of ARSB might contribute to the pathophysiology of CF. Previous cell-based experiments had demonstrated that when the deficiency of the cystic fibrosis transmembrane regulator (CFTR) was corrected in bronchial epithelial cells, the ARSB activity increased significantly. De-identified, citrated blood samples were collected from 16 children with cystic fibrosis and 31 control subjects, seen in the Pediatric Clinic at Rush University Medical Center. Polymorphonuclear (PMN) and mononuclear cell (MC) populations were separated by density gradient, and blinded determinations of ARSB activity were performed using the exogenous substrate 4-methylumbilliferyl sulfate. Interleukin-6 was measured in the plasma samples by ELISA. ARSB activity was significantly less in the PMN and MC from the CF patients than controls (p<0.0001, unpaired t-test, two-tailed). Interleukin-6 levels in plasma were significantly greater in the CF population (p<0.001). Mean age, age range, and male:female ratio of CF patients and controls were similar, and no association of ARSB activity with age, gender, or CFTR genotype was evident. Since recombinant human ARSB is used successfully for replacement therapy in Mucopolysaccharidosis VI, it may be useful to restore ARSB activity to normal levels and increase degradation of sulfated GAGs in CF patients. PMID:22550062

Sharma, Girish; Burke, Jenifer; Bhattacharyya, Sumit; Sharma, Neha; Katyal, Shivani; Park, R. Lucy; Tobacman, Joanne

2013-01-01

237

Effects of Laparoscopic and Conventional (Open) Cholecystectomy on Human Leukocyte Antigen-DR Expression in Peripheral Blood Monocytes: Correlations with Immunologic StatusRID=\\  

Microsoft Academic Search

. It is well known that surgery\\u000a \\u000a significantly decreases immune responses. Laparoscopic cholecystectomy\\u000a \\u000a (LC) is a “miniinvasive” surgical procedure; and on the basis of\\u000a \\u000a this consideration we have investigated if and how the immune response\\u000a \\u000a is modified in patients after laparoscopic cholecystectomy compared to\\u000a \\u000a patients who underwent open cholecystectomy. Immune activity\\u000a \\u000a [neutrophils, total lymphocytes count, lymphocytes subpopulations,\\u000a \\u000a human leukocyte antigen-DR

Francesco Carlei; Mario Schietroma; Giovanni Cianca; Alberto Risetti; Sandro Mattucci; Gerard Ngome Enang; Mario Simi

1999-01-01

238

Gene expression profile in human leukocytes.  

PubMed

Leukocytes are classified as myelocytic or lymphocytic, and each class of leukocytes consists of several types of cells that have different phenotypes and different roles. To define the gene expression in these cells, we have performed serial analysis of gene expression (SAGE) using human leukocytes and have provided the gene database for these cells not only at the resting stage but also at the activated stage. A total of 709,990 tags from 17 libraries were analyzed for the manifestation of gene expression profiles in various types of human leukocytes. Types of leukocytes analyzed were as follows: peripheral blood monocytes, colony-stimulating factor-induced macrophages, monocyte-derived immature dendritic cells, mature/activated dendritic cells, granulocytes, natural killer (NK) cells, resting B cells, activated B cells, naive T cells, CCR4(-) memory T cells (resting T(H)1 cells), CCR4(+) memory T cells (resting T(H)2 cells), activated T(H)1 cells, and activated T(H)2 cells. Among 38,961 distinct tags that appeared more than once in the combined total libraries, 27,323 tags were found to represent unique genes in certain type(s) of leukocytes. Using probability (P) and hierarchical clustering analysis, we identified the genes selectively expressed in each type of leukocytes. Identification of the genes specifically expressed in different types of leukocytes provides not only a novel molecular signature to define different subsets of resting and activated cells but also contributes to further understanding of the biologic function of leukocytes in the host defense system. PMID:12522010

Hashimoto, Shin-ichi; Nagai, Shigenori; Sese, Jun; Suzuki, Takuji; Obata, Aya; Sato, Taku; Toyoda, Nobuaki; Dong, Hong-Yan; Kurachi, Makoto; Nagahata, Tomoyuki; Shizuno, Ken-ichi; Morishita, Shinichi; Matsushima, Kouji

2003-05-01

239

Feature selection and classification of leukocytes using random forest.  

PubMed

In automatic segmentation of leukocytes from the complex morphological background of tissue section images, a vast number of artifacts/noise are also extracted causing large amount of multivariate data generation. This multivariate data degrades the performance of a classifier to discriminate between leukocytes and artifacts/noise. However, the selection of prominent features plays an important role in reducing the computational complexity and increasing the performance of the classifier as compared to a high-dimensional features space. Therefore, this paper introduces a novel Gini importance-based binary random forest feature selection method. Moreover, the random forest classifier is used to classify the extracted objects into artifacts, mononuclear cells, and polymorphonuclear cells. The experimental results establish that the proposed method effectively eliminates the irrelevant features, maintaining the high classification accuracy as compared to other feature reduction methods. PMID:25284218

Saraswat, Mukesh; Arya, K V

2014-12-01

240

Leukocyte trafficking in experimental autoimmune uveitis in vivo  

Microsoft Academic Search

Leukocyte trafficking from blood into tissue is a fundamental process in immune surveil- lance and the immune response to stimuli. Experi- mental autoimmune uveitis (EAU) is an animal model for posterior uveitis and is mediated by T lymphocytes and macrophages that infiltrate the posterior segment of the eye. To analyze leukocyte migration into retinal tissue during the course of EAU,

Adrian Parnaby-Price; Miles R. Stanford; John Biggerstaff; Lucy Howe; Roy A. Whiston; John Marshall; Graham R. Wallace

1998-01-01

241

Leukocyte adherence to venular endothelium during ischemia-reperfusion.  

PubMed

Xanthine oxidase-derived oxidants and leukocytes have been implicated in the microvascular injury associated with reperfusion of ischemic intestine. The objective of this study was to determine whether xanthine oxidase-derived oxidants play a role in the leukocyte-microvascular interactions initiated by ischemia-reperfusion. Adherence and extravasation of leukocytes were monitored in cat mesenteric venules subjected to 1 h of ischemia (blood flow reduced to 20% of control) and reperfusion. Leukocyte rolling velocity, vessel diameter, and red cell velocity were also measured in control (untreated) animals and in animals pretreated with either allopurinol or superoxide dismutase. The responses of venular blood flow, wall shear rate, and leukocyte rolling velocity to ischemia and reperfusion did not differ between the three experimental groups. In control animals, 1 h of ischemia was associated with significant adherence and extravasation of leukocytes with reperfusion greatly enhancing these responses. Allopurinol treatment did not alter the responses to ischemia per se, yet it largely prevented the further increment in adherence and extravasation associated with reperfusion. Superoxide dismutase treatment attenuated the leukocyte responses elicited by both ischemia and reperfusion. Our observations that both allopurinol and superoxide dismutase attenuate reperfusion-induced leukocyte adherence and extravasation are consistent with the hypothesis that xanthine oxidase-derived oxidants initiate the leukocyte infiltration induced by reperfusion of ischemic intestine. PMID:2596604

Granger, D N; Benoit, J N; Suzuki, M; Grisham, M B

1989-11-01

242

Oxygen radical production by avian leukocytes.  

PubMed Central

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

Conlon, P; Smith, D; Gowlett, T

1991-01-01

243

Release of Infectious Epstein-Barr Virus by Transformed Marmoset Leukocytes  

Microsoft Academic Search

Marmoset blood leukocytes transformed in vitro by Epstein-Barr virus regularly release extracellular infectious Epstein-Barr virus with high titers of transforming activity. By comparison, human umbilical cord leukocytes and adult human leukocytes transformed by Epstein-Barr virus release either no extracellular infectious virus or small amounts, irregularly.

George Miller; Muriel Lipman

1973-01-01

244

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

Microsoft Academic Search

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

Pia Kivisäkk; Grahame Kidd; Richard M. Ransohoff

2003-01-01

245

Size referenced electronic leukocyte counting threshold and lysed leukocyte size distribution of common domestic animal species.  

PubMed

Using a single channel electronic cell counter and attached particle size analyzer, leukocyte size distribution histograms were determined on canine, feline, bovine, and equine blood diluted with chloride-based diluent and treated with a conventional stromatolysin. Histograms were usually unimodal, but a few were bimodal. Mean values for mean lysed leukocyte particle volume were 49.2, 51.1, 55.4, and 65.0 fl for canine, feline, equine, and bovine blood, respectively. From inspection of histograms, a lower threshold of 30 fl referenced to latex spheres was interpreted to be appropriate for counting leukocytes of these four species simultaneously. Debris below the threshold was seen in many samples and was usually separated from the leukocyte population by a valley touching the histogram baseline at the threshold channel. Debris resulted in a visually detectable threshold failure by extending considerably into the leukocyte size range in 9% of feline, 9% of canine, and 7% of bovine samples. It is recommended that careful establishment of the lower counting threshold will minimize frequency and severity of leukocyte count error associated with failure to exclude debris. PMID:3455085

Weiser, M G

1987-11-01

246

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

PubMed Central

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

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

2009-01-01

247

Electrophoretic detection of protein p53 in human leukocytes  

SciTech Connect

The authors have found an acid-soluble protein with mol. wt. of about 53 kD in peripheral blood leukocytes of persons with Down's syndrome. It was present in different quantities in all 20 patients tested, but was virtually not discovered in 12 healthy blood donors. This paper determines the possible identity of this protein with protein p53 from mouse ascites carcinoma by comparing their electrophoretic mobilities, because the accuracy of electrophoretic determination of the molecular weight of proteins is not sufficient to identify them. The paper also describes experiments to detect a protein with electrophoretic mobility identical with that of a protein in the leukocytes of patients with Down's syndrome in leukocytes of patients with leukemia. To discover if protein p53 is involved in cell proliferation, the protein composition of leukocytes from healthy blood donors, cultured in the presence and absence of phytohemagglutinin (PHA), was compared. Increased incorporation of H 3-thymidine by leukocytes of patients with Down's syndrome is explained by the presence of a population of immature leukocytes actively synthesizing DNA in the peripheral blood of these patients, and this can also explain the presence of protein p53 in the leukocytes of these patients.

Paponov, V.D.; Kupsik, E.G.; Shcheglova, E.G.; Yarullin, N.N.

1986-01-01

248

Blood  

MedlinePLUS

... mysterious, life-sustaining fluid called blood. What Is Blood and What Does It Do? Two types of ... mixture of blood cells and plasma. Continue Red Blood Cells Red blood cells (RBCs, and also called ...

249

Blood  

MedlinePLUS

... solid part of your blood contains red blood cells, white blood cells, and platelets. Red blood cells deliver oxygen from your lungs to your tissues and organs. White blood cells fight infection and are part of your body's ...

250

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

PubMed Central

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.

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

2014-01-01

251

Nano-surgery at the leukocyte-endothelial docking site.  

PubMed

The endothelium has an important role in controlling the extravasation of leukocytes from blood to tissues. Endothelial permeability for leukocytes is influenced by transmembrane proteins that control inter-endothelial adhesion, as well as steps of the leukocyte transmigration process. In a cascade consisting of leukocyte rolling, adhesion, firm adhesion, and diapedesis, a new step was recently introduced, the formation of a docking structure or "transmigratory cup." Both terms describe a structure formed by endothelial pseudopods embracing the leukocyte. It has been found associated with both para- and transcellular diapedesis. The aim of this study was to characterize the leukocyte-endothelial contact area in terms of morphology and cell mechanics to investigate how the endothelial cytoskeleton reorganizes to engulf the leukocyte. We used atomic force microscopy (AFM) to selectively remove the leukocyte and then analyze the underlying cell at this specific spot. Firmly attached leukocytes could be removed by AFM nanomanipulation. In few cases, this exposed 8-12 microm wide and 1 microm deep footprints, representing the cup-like docking structure. Some of them were located near endothelial cell junctions. The interaction area did not exhibit significant alterations neither morphologically nor mechanically as compared to the surrounding cell surface. In conclusion, the endothelial invagination is formed without a net depolymerization of f-actin, as endothelial softening at the site of adhesion does not seem to be involved. Moreover, there were no cases of phagocytotic engulfment, but instead the formation of a transmigratory channel could be observed. PMID:18094992

Riethmuller, Christoph; Nasdala, Ines; Vestweber, Dietmar

2008-04-01

252

Leukocyte adhesion in local versus hemorrhage-induced ischemia.  

PubMed

The objective of this study was to compare the leukocyte-endothelial cell adhesive interactions elicited in postcapillary venules by either local ischemia-reperfusion or hemorrhage-reperfusion. Leukocyte rolling, adherence, and emigration were monitored in cat mesenteric venules exposed to an 85% reduction in blood flow (induced by either hemorrhage or local restriction of arterial inflow) for 1 h, followed by 1 h reperfusion. Leukocyte-endothelial cell interactions, venular diameter, and red blood cell velocity were measured during baseline, ischemia, and reperfusion periods. Both local and hemorrhage-induced ischemia reperfusion caused a reduction in leukocyte rolling velocity and increases in leukocyte adherence and emigration. Quantitatively, the adherence and emigration responses in both ischemia models were nearly identical. However, the two models differed in their response to immunoneutralization of the leukocyte adhesion glycoprotein CD11/CD18 with monoclonal antibody (MAb) IB4. The MAb had a more profound effect in attenuating leukocyte adherence and emigration in the local ischemia model. These results indicate that different factors may contribute to leukocyte-endothelial cell adhesive interactions observed in local vs. systemic models of ischemia-reperfusion. PMID:1415607

Perry, M A; Granger, D N

1992-09-01

253

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

PubMed Central

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

1979-01-01

254

Serum opsonin, bacteria, and polymorphonuclear leukocyte interactions in subacute bacterial endocarditis  

PubMed Central

The effect of anti-?-globulin factors on 7S ?-globulin opsonins from patients with subacute bacterial endocarditis has been examined with a quantitative in vitro phagocytosis system. Human anti-?-globulin factors from patients with subacute bacterial endocarditis and rheumatoid arthritis inhibited the opsonic action of 7S ?-globulin specifically bound to bacteria. A similar antiopsonic effect was obtained with rabbit antiserum to human ?G globulin. The antiopsonic effect of anti-?-globulin factors did not correlate with their ability to potentiate agglutination of bacteria by 7S antibody. Competition was demonstrated between the antiopsonic effect of anti-?-globulin factors and the phagocytosis-promoting action of heat-labile serum factors containing hemolytically active complement. PMID:5645856

Messner, Ronald P.; Laxdal, Throstur; Quie, Paul G.; Williams, Ralph C.

1968-01-01

255

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

E-print Network

will determine if differences in a specific neutrophil function occur with age . Combining information from this study with that available from other studies will be helpful in further elucidating age-related change in PMNs. An age-related comparison of 02... will determine if differences in a specific neutrophil function occur with age . Combining information from this study with that available from other studies will be helpful in further elucidating age-related change in PMNs. An age-related comparison of 02...

Hanson, Thomas Dale

2012-06-07

256

Precipitated immune complexes of IgM induce the generation of reactive oxygen species by rabbit polymorphonuclear leucocytes  

Microsoft Academic Search

We report that immune complexes of IgM (ICIgM) antibodies and ovalbumin in the form of a precipitate from the equivalence zone induce the generation of reactive oxygen species by rabbit blood polymorphonuclear leucocytes (PMN), as measured by the chemilu- minescence (CL) production in the presence of luminol. The kinetics of CL generation induced by ICIgM is quite different from that

Y. M. Lucisano; A. M. de-Mello; E. R. Vasques; B. Mantovani

1998-01-01

257

Cyclic nucleotide changes induced in human leukocytes by a product of axenically grown Entamoeba histolytica that inhibits human monocyte locomotion  

Microsoft Academic Search

Pulse exposure of human mononuclear phagocytes to the monocyte locomotion-inhibitory factor produced byEntamoeba histolytica (i.e., the 369- to 765-Da chromatographic fraction obtained from the supernatant fluid of axenically grownE. histolytica) led to a swift increase in the intracellular concentration of adenosine 3':5' cyclic monophosphate (cAMP). A weaker response was observed in human polymorphonuclear leukocytes, the locomotion of which, however, is

G. Rico; O. Díaz-Guerra; R. R. Kretschmer

1995-01-01

258

Role of tumor necrosis factor ? release and leukocyte margination in indomethacin-induced gastric injury in rats  

Microsoft Academic Search

Background\\/Aims: Several studies have shown that polymorphonuclear neutrophil leukocyte (PMN) margination is an early and critical event in the pathogenesis of gastric mucosal injury caused by nonsteroidal anti-inflammatory drugs. Tumor necrosis factor (TNF) ? is a proinflammatory cytokine that causes PMN margination by up-regulating expression of adhesion molecules on both PMN and endothelial cells. This study investigated whether substances that

Luca Santucci; Stefano Fiorucci; Francesco Maria Di Matteo; Antonio Morelli

1995-01-01

259

Influence of plasma factors and drugs on in vitro leukocyte adhesiveness.  

PubMed

Leukocyte adhesiveness was studied by measuring the retention of leukocytes in glass bead columns after the passage of heparinized whole blood or leukocyte suspensions. Leukocytes adhered readily to glass beads and independently of divalent cations when proteins were absent. Addition of plasma inhibited leukocyte adherence which became increasingly dependent on divalent cations. In conflict with results reported from nylon wool column experiments, we could not demonstrate a leukocyte adherence augmenting effect of zymosan activated plasma or plasma from patients with inflammatory diseases or patients undergoing hemodialysis. Ethanol and lidocaine gave a dose response inhibition of leukocyte adherence. The conflicting results obtained with the nylon wool and the glass head column techniques in conditions with "augmented adherence" may be due to retention of leukocyte aggregates in the nylon wool columns. PMID:6782662

Hopen, G

1980-01-01

260

Singlet oxygen inactivates fibrinogen, factor V, factor VIII, factor X, and platelet aggregation of human blood.  

PubMed

Activated polymorphonuclear leukocytes participate in hemostasis. These phagocytes generate up to 5 mmol/l of oxidants of the HOCl- and chloramine-type. The present study shows, for the first time, that physiological concentrations of NaOCl or chloramines act as anticoagulants in human plasma. Prothrombin time, activated partial thromboplastin time, and thrombin time at chloramine concentrations greater than 1 mmol/l are prolonged proportional to the oxidant concentration. Plasmatic coagulation factors sensible to oxidation are fibrinogen, factor V, factor VIII, and factor X with a 50% effective dose of 2-3 mmol/l NaOCl or taurine-chloramine. Chloramines or chloramine-like agents (e.g., chloramine T(R) or vancomycin) also inactivate platelet aggregation (in whole blood or platelet-rich plasma) at an 50% effective dose of about 1.0 mmol. This irreversible oxidation of the hemostasis components is inhibited by addition of methionine, cysteine, ascorbic acid, or azide in 10-fold molar excess prior to oxidation. The oxy-radical inhibitors mannitol, superoxide dismutase, or catalase do not antagonize the action of NaOCl or chloramines. Therefore, the oxidant here involved has reaction characteristics of singlet oxygen (1O(2)), a nonradical, excited (i.e., light-emitting) oxidant. The hemostasis factors sensible to oxidation might dispose of oxidizable, for their function critical, methionine or cysteine residues. In conclusion, blood coagulation factors I, V, VIII, X and thrombocytes are sensible to nonradical oxidants of activated phagocytes. Via 1O(2) generation, polymorphonuclear leukocytes can generate a local pericellular zone of anticoagulation. The data suggest that the cell signal 1O(2) in physiological amounts is an antithrombotic agent. PMID:10704657

Stief, T W; Kurz, J; Doss, M O; Fareed, J

2000-03-15

261

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

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

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

2014-01-01

262

Uptake of indium-111-labeled leukocytes by brain metastasis  

SciTech Connect

Uptake of indium-labeled leukocytes was seen in two cases of histologically proven brain metastasis. In one, this led to misdiagnosis of the lesion as an abscess. On histological evaluation, a large number of white blood cells or macrophages was seen at the neoplastic sites. Reasons for leukocyte accumulation around metastatic brain neoplasms are discussed. In contrast to the current reports that indium-labeled leukocyte scans can differentiate intracranial infection from tumor, these cases demonstrate their lack of specificity in the detection of brain abscess.

Balachandran, S.; Husain, M.M.; Adametz, J.R.; Pallin, J.S.; Angtuaco, T.L.; Boyd, C.M.

1987-04-01

263

Polymorphonuclear neutrophil in brain parenchyma after experimental intracerebral hemorrhage.  

PubMed

Polymorphonuclear neutrophils (PMNs) infiltration into brain parenchyma after cerebrovascular accidents is viewed as a key component of secondary brain injury. Interestingly, a recent study of ischemic stroke suggests that after ischemic stroke, PMNs do not enter brain parenchyma and as such may cause no harm to the brain. Thus, the present study was designed to determine PMNs' behavior after intracerebral hemorrhage (ICH). Using the autologous blood injection model of ICH in rats and immunohistochemistry for PMNs and vascular components, we evaluated the temporal and spatial PMNs distribution in the ICH-affected brain. We found that, similar to ischemia, there is a robust increase in presence of PMNs in the ICH-injured tissue that lasts for at least 1 to 2 weeks. However, in contrast to what was suggested for ischemia, besides PMNs that stay in association with the vasculature, after ICH, we found abundance of intraparenchymal PMNs (with no obvious association with vessels) in the ICH core and hematoma border, especially between 1 and 7 days after the ictus. Interestingly, the increased presence of intraparenchymal PMNs after ICH coincided with the massive loss of microvascular integrity, suggesting vascular disruption as a potential cause of PMNs presence in the brain parenchyma. Our study indicates that in contrast to ischemic stroke, after ICH, PMNs target not only vascular compartment but also brain parenchyma in the affected brain. As such, it is possible that the pathogenic role and therapeutic implications of targeting PMNs after ICH could be different from these after ischemic stroke. Our work suggests the needs for more studies addressing the role of PMNs in ICH. PMID:24696130

Zhao, Xiurong; Sun, Guanghua; Zhang, Han; Ting, Shun-Ming; Song, Shen; Gonzales, Nicole; Aronowski, Jaroslaw

2014-10-01

264

Biomechanics of leukocyte rolling  

PubMed Central

Leukocyte rolling on endothelial cells and other P-selectin substrates is mediated by P-selectin binding to P-selectin glycoprotein ligand-1 expressed on the tips of leukocyte microvilli. Leukocyte rolling is a result of rapid, yet balanced formation and dissociation of selectin-ligand bonds in the presence of hydrodynamic shear forces. The hydrodynamic forces acting on the bonds may either increase (catch bonds) or decrease (slip-bonds) their lifetimes. The force-dependent ‘catch-slip’ bond kinetics are explained using the ‘two pathway model’ for bond dissociation. Both the ‘sliding-rebinding’ and the ‘allosteric’ mechanisms attribute ‘catch-slip’ bond behavior to the force-induced conformational changes in the lectin-EGF domain hinge of selectins. Below a threshold shear stress, selectins cannot mediate rolling. This ‘shear-threshold’ phenomenon is a consequence of shear-enhanced tethering and catch-bond enhanced rolling. Quantitative dynamic footprinting microscopy has revealed that leukocytes rolling at venular shear stresses (> 0.6 Pa) undergo cellular deformation (large footprint) and form long tethers. The hydrodynamic shear force and torque acting on the rolling cell are thought to be synergistically balanced by the forces acting on tethers and stressed microvilli, however, their relative contribution remains to be determined. Thus, improvement beyond the current understanding requires in silico models that can predict both cellular and microvillus deformation and experiments that allow measurement of forces acting on individual microvilli and tethers. PMID:21515934

Sundd, Prithu; Pospieszalska, Maria K.; Cheung, Luthur Siu-Lun; Konstantopoulos, Konstantinos; Ley, Klaus

2011-01-01

265

Degradation of cartilage proteoglycan by human leukocyte granule neutral proteases--a model of joint injury. I. Penetration of enzyme into rabbit articular cartilage and release of 35SO4-labeled material from the tissue.  

PubMed Central

The present work was undertaken to explore the effect of two purified neutral proteases derived from human peripheral blood polymorphonuclear leukocytes (PMN) on articular cartilage as a model of joint injury. Human leukocyte elastase and chymotrypsin-like enzyme, purified by affinity chromatography, released 32SO4 from labeled rabbit articular cartilage slices in vitro. Release of isotope was initially delayed, suggesting that either a lag in enzyme penetration occurs or that size of degradation fragments is a limiting factor in diffusion of label out of the tissue. The release of 35SO4 was inhibited by preincubation of elastase and chymotrypsin-like enzyme with human alpha 1-anti-trypsin, or with their specific chloromethyl ketone inactivators, and the action of elastase was also inhibited by a monospecific antiserum to PMN elastase, freed of major serum proteinase inhibitors. Immunohistochemical staining procedures revealed the presence of PMN elastase inside the matrix of cartilage slices after a 20-min exposure of tissue to either the pure enzyme or crude PMN granule extract. Serum alpha 1-antitrypsin failed to penetrate into the cartilage slices under identical in vitro conditions. In association with the results reported in the accompanying paper, these findings suggest a model of cartilage matrix degradation by PMN neutral proteases in which local protease-antiprotease imbalance, coupled with different rates of penetration of protease and antiprotease into target tissue, plays a key role in accounting for matrix damage. Images PMID:129482

Janoff, A; Feinstein, G; Malemud, C J; Elias, J M

1976-01-01

266

The onset of polymorphonuclear leucocyte membrane-stimulated metabolic activity.  

PubMed Central

The time course of polymorphonuclear leucocyte oxidative metabolism following membrane stimulation by four different agents was examined using the techniques of luminol- and lucigenin-dependent chemiluminescence. After addition of opsonized zymosan, phorbol myristate acetate or digitonin to polymorphonuclear leucocytes there was a lag period of between 35 and 55 sec before the onset of chemiluminiscence. In contrast, after addition of the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (FMLP), the lag period before chemiluminescence was less than 19 sec. Luminol-dependent chemiluminescence was reduced by superoxide dismutase and almost abolished by sodium azide. The inhibitory effect of the latter was less marked when using FMLP. Lucigenin-dependent chemiluminescence was inhibited by superoxide dismutase and enhanced by sodium azide. Cytochalasin B reduced zymosan and digitonin stimulated chemiluminescence but increased FMLP stimulated chemiluminescence. The results of the onset of polymorphonuclear leucocyte metabolic activity using other techniques. PMID:7275176

Williams, A J; Cole, P J

1981-01-01

267

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

NASA Astrophysics Data System (ADS)

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

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

2005-10-01

268

Rheological modelling of leukocytes.  

PubMed

A three-layer Newtonian model is investigated using a combined Eulerian-Lagrangian computational method to describe the dynamic behaviour of leukocytes. The model, composed of a cell membrane (outer layer), cytoplasm (middle layer) and nucleus (inner layer), can better describe the recovery characteristics because large viscosity and capillarity differences between layers are considered, and both Newtonian and seemingly non-Newtonian behaviours reported in the literature can be reproduced. It is found that, to describe adequately the various rheological characteristics of leukocytes, the presence of the highly viscous nucleus and its deformation/recovery, as well as the surface energy stored in the fluid interfaces, are critical. Photographs from pipette experiments using a fluorescent technique confirm the theoretical finding of the important role played by the nucleus in cell deformation. PMID:9684470

Tran-Son-Tay, R; Kan, H C; Udaykumar, H S; Damay, E; Shyy, W

1998-03-01

269

Leukocyte labeling with technetium-99m tin colloids.  

PubMed

Triple density gradients of metrizamide in plasma (MP) were used to characterize label distribution in human leukocyte preparations incubated with 99mTc tin colloids. Less than 50% of the cell-associated radioactivity was specifically bound to leukocytes when heparinized blood was rotated with stannous fluoride colloid ([Tc]SFC). Labeling efficiency in leukocyte rich plasma (LRP) averaged 44%, of which greater than 90% was specifically bound to leukocytes. MP-gradient analysis also revealed that leukocyte labeling did not occur with stannous chloride colloid, nor when citrate was present during rotation with [Tc]SFC. When citrate was added after labeling to "solubilize" unbound [Tc]SFC, radiocolloid was removed from the leukocytes, indicating that the mechanism of [Tc]SFC labeling is adherence rather than phagocytosis. Technetium-labeled neutrophils exhibited normal in vitro chemotaxis and no lung uptake in vivo. Technetium-labeled mononuclear leukocytes, on the other hand, exhibited prolonged lung transit in vivo. Neither [Tc]SFC cell preparation showed signs of in vivo reoxidation to pertechnetate. PMID:3625299

Mock, B H; English, D

1987-09-01

270

Role of superoxide dismutase and catalase as determinants of pathogenicity of Nocardia asteroides: importance in resistance to microbicidal activities of human polymorphonuclear neutrophils.  

PubMed Central

The roles of nocardial superoxide dismutase (SOD) and catalase in the resistance of Nocardia asteroides to the microbicidal properties of human polymorphonuclear leukocytes were determined in vitro. The neutrophils killed ca. 80% of the cells of the less virulent N. asteroides 10905 and ca. 50% of the log phase of the more virulent N. asteroides GUH-2 after 180 min of incubation. These phagocytes were not able to kill early-stationary-phase cells of strain GUH-2 that contained 10 times more intracytoplasmic catalase than log-phase cells of the same culture. However, the polymorphonuclear leukocytes were able to kill more than 50% of the cells of early-stationary-phase strain GUH-2 after treatment with purified antibody specific for surface-associated SOD. No killing was observed when the bacteria were treated with normal rabbit immunoglobulin G or with serum obtained from rabbits immunized against whole nocardial cells (containing little or no activity against SOD). These phagocytes killed more than 99% of Listeria monocytogenes used as a control. Chlorpromazine-treated polymorphonuclear leukocytes killed L. monocytogenes (70%) but they were not able to kill antibody-treated cells of N. asteroides GUH-2. Exogenously added SOD partially protected strain 10905, which lacked surface-associated enzyme, but it had no effect on the killing of strain GUH-2, which already possessed significant amounts of surface-bound SOD. In contrast, catalase added to the nocardiae provided almost complete protection to the log-phase cells of strain GUH-2, but strain 10905 was only partially protected. SOD combined with catalase had additive activity which completely protected the cells of strain 10905. A mutant of N. asteroides GUH-2 (SCII-C) is more virulent during the log phase than is the parental strain. This mutant contained at least 7 times more catalase at this stage of growth than did the parent. No other differences between these two strains were observed during the log phase. In sharp contrast to those of the parent, log-phase cells of this high-catalase mutant were not killed by polymorphonuclear phagocytes. These data indicate a role for both SOD and catalase in the resistance of Nocardia spp. to human neutrophils, and they represent at least two factors associated with virulence. PMID:3880721

Beaman, B L; Black, C M; Doughty, F; Beaman, L

1985-01-01

271

In Vivo Leukocyte Changes Induced by Escherichia coli Subtilase Cytotoxin?  

PubMed Central

Subtilase cytotoxin (SubAB) is the prototype of a new family of AB5 cytotoxins produced by Shiga-toxigenic Escherichia coli. Its cytotoxicity is due to its capacity to enter cells and specifically cleave the essential endoplasmic reticulum chaperone BiP. Previous studies have shown that intraperitoneal injection of mice with purified SubAB causes a pathology that overlaps with that seen in human cases of hemolytic-uremic syndrome, as well as dramatic splenic atrophy, suggesting that leukocytes are targeted. Here we investigated SubAB-induced leukocyte changes in the peritoneal cavity, blood, and spleen. After intraperitoneal injection, SubAB bound peritoneal leukocytes (including T and B lymphocytes, neutrophils, and macrophages). SubAB elicited marked leukocytosis, which peaked at 24 h, and increased neutrophil activation in the blood and peritoneal cavity. It also induced a marked redistribution of leukocytes among the three compartments: increases in leukocyte subpopulations in the blood and peritoneal cavity coincided with a significant decline in splenic cells. SubAB treatment also elicited significant increases in the apoptosis rates of CD4+ T cells, B lymphocytes, and macrophages. These findings indicate that apart from direct cytotoxic effects, SubAB interacts with cellular components of both the innate and the adaptive arm of the immune system, with potential consequences for disease pathogenesis. PMID:21282417

Wang, Hui; Paton, Adrienne W.; McColl, Shaun R.; Paton, James C.

2011-01-01

272

In vitro comparison of HMPAO and gentisic acid for labelling leukocytes with 99mTc.  

PubMed

Leukocytes can be labelled with 99mTc using HMPAO and gentisic acid methods. We compared the two methods with respect to labelling efficiency on mixed leukocytes and isolated polymorphonuclear (PMN) and mononuclear (MN) cells, and the in vitro stability of the label. HMPAO produced approximately 70% labelling efficiency on mixed or PMN cells and the label was stable in saline or plasma. Labelling efficiency on MN was only 14% and was less stable. Gentisic acid produced a labelling efficiency of 52% on PMN and 35% on MN; both were stable in saline but less stable in plasma. In conclusion, HMPAO produces higher labelling efficiency and the label shows greater in vitro stability in plasma. However, gentisic acid is much less expensive to use, allows labelling of MN cells, and should result in more favourable microdosimetry. Preliminary clinical results suggest that gentisic acid is equivalent to HMPAO but has the advantage of being much cheaper. PMID:2161770

Ecclestone, M; Proulx, A; Ballinger, J R; Gerson, B; Reid, R H; Gulenchyn, K Y

1990-01-01

273

In vitro effects of 1,25-dihydroxyvitamin D3 on interferon-gamma and tumor necrosis factor-alpha secretion by blood leukocytes from young and adult cattle vaccinated with Mycobacterium bovis BCG.  

PubMed

Interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) are critical in the development of an effective immune response. Vitamin D, essential in short-term calcium homeostasis and recently shown to modulate proliferation and function of blood mononuclear cells from adult dairy cattle, may be an effective modulator of the calf's immune system. Effects of antigen sensitization and 1,25-dihydroxyvitamin D3[1,25-(OH)2D3] on cytokine secretion by cells from calves vaccinated with Bacille Calmette-Guérin (BCG) were examined. One-week-old dairy calves (n = 6) and yearling heifers (n = 4) were vaccinated concurrently with BCG and boosted six weeks later. Ten weeks after primary vaccination, cells from vaccinated calves and adults, and nonvaccinated, age-matched calves (n = 4) were evaluated in vitro for their capacity to produce IFN-gamma and TNF-alpha. Cells were stimulated with pokeweed mitogen (PWM) or recall antigen [Mycobacterium bovis-derived purified protein derivative (PPD)] in the presence of 0, 0.1, 1.0, and 10 nM of 1,25-(OH)2D3 for 20, 44, and 68 hours, respectively. IFN-gamma and TNF-alpha concentrations in culture supernatants harvested at these times were quantified by enzyme-linked immunosorbent assay (ELISA). PPD-induced IFN-gamma and TNF-alpha responses of cells from vaccinated calves and adults were greater than responses of autologous unstimulated cells. In contrast, PPD-specific responses of calf and adult cells collected immediately before primary vaccination were substantially lower and comparable to responses in resting (i.e., unstimulated) cultures. At ten weeks, the PPD-specific response of vaccinates exceeded the response of nonvaccinated calves; however, responses of vaccinated calves were more vigorous than corresponding responses of vaccinated adults. Incubation period also influenced the magnitude of both IFN-gamma and TNF-alpha, responses in PPD- and PWM-stimulated cultures. Effects of 1,25-(OH)2D3 on antigen-induced secretion of IFN-gamma and TNF-alpha were marginal. Only IFN-gamma responses of vaccinated adults were affected by 1,25-(OH)2D3. Vitamin D caused a concentration-dependent decrease in IFN-gamma response and an increase in TNF-alpha response in PWM-stimulated cultures. These results indicate that animal maturity (i.e., age) and antigenic experience affect IFN-gamma and TNF-alpha secretion by bovine leukocytes and suggest that 1,25-(OH)2D3 can alter secretion of both cytokines under specific conditions of culture. PMID:12951895

Nonnecke, Brian J; Waters, W Ray; Foote, Monica R; Horst, Ronald L; Fowler, Mike A; Miller, Bill L

2003-07-01

274

Neutrophil-derived oxidants promote leukocyte adherence in postcapillary venules.  

PubMed

The objective of this study was to determine whether hydrogen peroxide (H2O2), hypochlorous acid (HOCl), and monochloramine (NH2Cl), at concentrations produced by activated neutrophils, promote leukocyte adherence to microvascular endothelium in post-capillary venules. Cat mesenteric venules (30-45 microns diameter) were examined using intravital video microscopy. Red blood cell velocity (VRBC), venular diameter (DV), and the number of adherent leukocytes (NWBC) were measured in postcapillary venules. Venular blood flow and wall shear rate (tau) were calculated from the measured values of VRBC and DV. Different concentrations (0.01-1.0 mM) of H2O2, HOCl, or NH2Cl were superfused on the mesentery. In some experiments, the contributions of the leukocyte adhesive glycoprotein CD11/CD18 and platelet-activating factor (PAF) in the oxidant-induced leukocyte adherence were determined using a CD18-specific antibody (IB4) and a PAF-receptor antagonist (WEB 2086), respectively. The results of our in vivo experiments indicate that H2O2 and NH2Cl, but not HOCl, promote leukocyte adhesion to venular endothelium. Incubation of isolated cat neutrophils with either NH2Cl or H2O2 resulted in activation of CD11/CD18, as assessed by flow cytometry. Although the leukocyte adhesion induced by both H2O2 and NH2Cl was associated with a reduction in venular wall shear rate, corresponding decrements in shear rate induced by partial occlusion of the mesenteric artery did not lead to similar levels of leukocyte adherence. The leukocyte adherence induced by H2O2 and NH2Cl was largely prevented by monoclonal antibody IB4, indicating that both oxidants promote leukocyte adherence via activation of CD11/CD18. The H2O2-induced, CD18-mediated leukocyte adherence appears to be elicited by PAF and by a direct effect of the oxidant on CD11/CD18 expression. The mechanism underlying the NH2Cl-induced leukocyte adherence remain unclear. PMID:1658575

Suzuki, M; Asako, H; Kubes, P; Jennings, S; Grisham, M B; Granger, D N

1991-09-01

275

Studies on PyrogenicFever Induced by Granulocytes- Free Leukocytes in Rabbits  

Microsoft Academic Search

The ability of rabbits' mononuclear cells to release a leukocytic pyrogen (LP) in vitro and its pyrogenecity were studied in the rabbits. Crude LP was obtained from granulocytes-free leukocytes as follows; the mixed cells of lymphocytes and monocytes had been separated on Ficoll-Conray gradients from whole blood of rabbits, and these cells were sensitized with bacterial lipopolysaccharide (LPS) in RPMI

Mariko FUJIWARA; Jun IWAMOTO; Nobu OHWATARI; Katsuhiko TSUCHIYA; Mitsuo KOSAKA; Tadamichi YANAGIand; Yoshiro TSUJI; Soeliadi HW

276

Local Oxidative and Nitrosative Stress Increases in the Microcirculation during Leukocytes-Endothelial Cell Interactions  

PubMed Central

Leukocyte-endothelial cell interactions and leukocyte activation are important factors for vascular diseases including nephropathy, retinopathy and angiopathy. In addition, endothelial cell dysfunction is reported in vascular disease condition. Endothelial dysfunction is characterized by increased superoxide (O2•?) production from endothelium and reduction in NO bioavailability. Experimental studies have suggested a possible role for leukocyte-endothelial cell interaction in the vessel NO and peroxynitrite levels and their role in vascular disorders in the arterial side of microcirculation. However, anti-adhesion therapies for preventing leukocyte-endothelial cell interaction related vascular disorders showed limited success. The endothelial dysfunction related changes in vessel NO and peroxynitrite levels, leukocyte-endothelial cell interaction and leukocyte activation are not completely understood in vascular disorders. The objective of this study was to investigate the role of endothelial dysfunction extent, leukocyte-endothelial interaction, leukocyte activation and superoxide dismutase therapy on the transport and interactions of NO, O2•? and peroxynitrite in the microcirculation. We developed a biotransport model of NO, O2•? and peroxynitrite in the arteriolar microcirculation and incorporated leukocytes-endothelial cell interactions. The concentration profiles of NO, O2•? and peroxynitrite within blood vessel and leukocytes are presented at multiple levels of endothelial oxidative stress with leukocyte activation and increased superoxide dismutase accounted for in certain cases. The results showed that the maximum concentrations of NO decreased ?0.6 fold, O2•? increased ?27 fold and peroxynitrite increased ?30 fold in the endothelial and smooth muscle region in severe oxidative stress condition as compared to that of normal physiologic conditions. The results show that the onset of endothelial oxidative stress can cause an increase in O2•? and peroxynitrite concentration in the lumen. The increased O2•? and peroxynitrite can cause leukocytes priming through peroxynitrite and leukocytes activation through secondary stimuli of O2•? in bloodstream without endothelial interaction. This finding supports that leukocyte rolling/adhesion and activation are independent events. PMID:22719984

Kar, Saptarshi; Kavdia, Mahendra

2012-01-01

277

Leucocyte Blood Picture in Ill Newborn Babies  

PubMed Central

Serial blood leucocyte counts were made on 35 ill preterm and term babies during the first 28 days of life. Ill babies with no clinical or bacteriological evidence of infection showed no changes in the blood leucocytes during the neonatal period when compared with normal babies. The changes in the blood leucocytes in babies with proven or suspected bacterial infection were an increase in the absolute values of polymorphonuclear neutrophils, an increase in the absolute values of immature neutrophils, a significant fall in polymorphonuclear neutrophils (below 1000/mm3) and in eosinophils (down to 0) in very ill babies, and toxic granulation of neutrophils. ImagesFIG. 3 PMID:4563923

Xanthou, Marietta

1972-01-01

278

CD31\\/PECAM-1 Is a Ligand for otv133 Integrin Involved in Adhesion of Leukocytes to Endothelium  

Microsoft Academic Search

To protect the body efficiently from infec- tious organisms, leukocytes circulate as nonadherent cells in the blood and lymph, and migrate as adherent cells into tissues. Circulating leukocytes in the blood have first to adhere to and then to cross the endothelial lining. CD31\\/PECAM-1 is an adhesion molecule ex- pressed by vascular endothelial cells, platelets, mono- cytes, neutrophils, and naive

Luca Piali; Philippe Hammel; Christoph Uherek; Felix Bachmann; Roland H. Gisler; Dominique Dunon; Beat A. Imhof

1995-01-01

279

Kinetics of reversible-sequestration of leukocytes by the isolated perfused rat lung  

SciTech Connect

The kinetics and morphology of sequestration and margination of rat leukocytes were studied using an isolated perfused and ventilated rat lung preparation. Whole rat blood, bone marrow suspension, or leukocyte suspensions, were used to perfuse the isolated rat lung. The lung was also perfused with latex particle suspensions and the passage of particles through the lung capillaries was studied. When a leukocyte suspension was perfused through the lung in the single-pass mode, the rate of sequestration decreased as more cells were perfused. In contrast, latex particles of a size comparable to that of leukocytes were totally stopped by the lung. When the leukocyte suspension was recirculated through the lung, cells were rapidly removed from circulation until a steady state was reached, after which no net removal of cells by the lung occurred. These results indicate that leukocytes are reversibly sequestered from circulation. The sequestered cells marginated and attached to the luminal surface of the endothelium of post-capillary venules and veins. A mathematical model was developed based on the assumption that the attachment and detachment of leukocytes to blood vessel walls follows first-order kinetics. The model correctly predicts the following characteristics of the system: (a) the kinetics of the sequestration of leukocytes by the lung; (b) the existence of a steady state when a suspension of leukocytes is recirculated through the lung; and (c) the independence of the fraction of cells remaining in circulation from the starting concentration for all values of starting concentration. (ERB)

Goliaei, B.

1980-08-01

280

Leukocyte response to eastern equine encephalomyelitis virus in a wild passerine bird.  

PubMed

Leukocyte counts are frequently used to assess the immunologic status of animals; however, few studies have directly looked at the predictive value of leukocyte counts and an animal's ability to respond to an infection with a pathogen. Understanding how an animal's leukocyte profile is altered by an active infection can assist with interpretation of leukocyte profiles in animals for which infection status is not known. In this study we examine the leukocyte counts of gray catbirds (Dumetella carolinensis) infected with eastern equine encephalomyelitis virus (EEEV). Blood smears were collected from infected catbirds on -4, 2, 5, and 14 days postinoculation (dpi) with EEEV, and from a corresponding uninfected control group, to monitor leukocyte counts. Although we found that preinfection leukocyte counts were not a reliable predictive of a catbird's viremia, we did find that infected catbirds exhibited significant hematologic changes in response to EEEV infection. We observed a significant drop in all subpopulations of leukocytes (i.e., lymphocytes, monocytes, and granulocytes) following infection. Lymphocytes and granulocytes still had not recovered to preinfection levels at 14 dpi. Uninfected catbirds also exhibited statistically significant changes in leukocyte counts, but this was due to a slight increase at 14 dpi and was not considered biologically relevant. Studies such as this can provide important information for field ecoimmunologists that use leukocyte counts to assess immunocompetence in free-living animals. PMID:24597116

Owen, J C; Cornelius, E A; Arsnoe, D A; Garvin, M C

2013-12-01

281

Red blood cells serve as intravascular carriers of myeloperoxidase.  

PubMed

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

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

282

Superoxide mediates reperfusion-induced leukocyte-endothelial cell interactions.  

PubMed

The objective of this study was to determine whether superoxide mediates the leukocyte-endothelial cell interactions elicited by reperfusion (reoxygenation) of ischemic (hypoxic) tissues. Mesenteric and intestinal blood flows were reduced to 20% of control for 1 h, followed by 1 h of reperfusion. Sixty minutes after reperfusion, red blood cell velocity (Vr), leukocyte rolling velocity (Vw), and the number of adherent leukocytes were measured in mesenteric venules. Then, either human superoxide dismutase (hSOD), hydrogen peroxide-inactivated hSOD, or MoAb IB4 (a monoclonal antibody against the leukocyte adhesion molecule CD18) was injected intravenously. Ten minutes later, repeat measurements were obtained and compared with pretreatment values. hSOD attenuated reperfusion-induced neutrophil adherence and increased Vw/Vr, an index of the fracture stress between leukocytes and endothelium. Peroxide-inactivated hSOD did not alter any parameter. MoAb IB4 attenuated reperfusion-induced adherence but did not alter Vw/Vr. In a correlate study, cultured bovine microvascular endothelium was exposed to 30 min of anoxia, followed by 60 min of reoxygenation. Cat neutrophils were added during reoxygenation. Reoxygenation-induced leukocyte adherence was attenuated by either hSOD or MoAb IB4 but not by inactivated hSOD. Adherence of phorbol 12-myristate 13-acetate-activated cat neutrophils to plastic was unaffected by hSOD or inactive hSOD, yet MoAb IB4 virtually abolished the response. These results indicate that superoxide mediates reperfusion-induced leukocyte adherence and that endothelial cells are required for this superoxide-mediated adherence. PMID:2556051

Suzuki, M; Inauen, W; Kvietys, P R; Grisham, M B; Meininger, C; Schelling, M E; Granger, H J; Granger, D N

1989-11-01

283

Role of platelet-activating factor in ischemia/reperfusion-induced leukocyte adherence.  

PubMed

The objective of this study was to determine whether platelet-activating factor (PAF) mediates the leukocyte-endothelial cell interactions elicited by ischemia/reperfusion. The rates of adherence and extravasation of leukocytes were monitored in cat mesenteric venules subjected to 60 min of ischemia (blood flow reduced to 20% of control) followed by 60 min of reperfusion. Leukocyte rolling velocity, red blood cell velocity, and vessel diameter were also measured. The experiments were performed in control (untreated) animals and in animals pretreated with one of two PAF receptor antagonists, i.e., BN 52021 or WEB 2086. The responses of venular blood flow, wall shear rate, and vessel diameter did not differ between the three groups. In the control group, 1 h of ischemia was associated with significant adherence and extravasation of leukocytes, with reperfusion greatly enhancing these responses. The rates of leukocyte adherence and extravasation during reperfusion were greatly attenuated by both PAF antagonists. Furthermore, the proportion of adherent leukocytes that ultimately extravasate during reperfusion was markedly reduced by WEB 2086. These results suggest that PAF plays an important role in mediating the adhesive interaction between circulating leukocytes and microvascular endothelium induced by ischemia/reperfusion and that the phospholipid promotes the leukocyte extravasation associated with ischemia/reperfusion. PMID:2166441

Kubes, P; Ibbotson, G; Russell, J; Wallace, J L; Granger, D N

1990-08-01

284

Quantitative reconstruction of leukocyte subsets using DNA methylation  

PubMed Central

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

2014-01-01

285

Morphology and density features of eosinophil leukocytes in eosinophilic pneumonia  

Microsoft Academic Search

Summary  We compared the morphological characteristics and density properties of eosinophil leukocytes obtained from the blood and bronchoalveolar lavage fluid of a 29-year-old patient with chronic eosinophil pneumonia during exacerbation. The lavage eosinophils were significantly increased in size when compared with blood cells (surface area: 208 ± 12 m2 versus 161 ± 13 m2). Moreover, eosinophils contained slightly more granules (23.4

C. Kroegel; H. Matthys; U. Costabel

1992-01-01

286

Effect of leptin on polymorphonuclear leucocyte functions in healthy subjects and haemodialysis patients  

PubMed Central

Background. Dysfunction of polymorphonuclear leucocytes (PMNLs) in end-stage renal disease (ESRD) patients contributes to a diminished immune defence. The serum levels of leptin are elevated in patients with ESRD. We analysed in vitro effects of leptin on PMNLs from healthy subjects (HS; n = 12) and haemodialysis (HD) patients (n = 15) before and after HD. Methods. PMNL oxidative burst and phagocytosis were tested by flow cytometry in whole blood. Chemotaxis of isolated PMNLs was assessed by the under-agarose method. To assess the involvement of leptin in PMNL signalling pathways, signal transduction inhibitors were used and the activity of intracellular kinases was investigated by western blotting, in vitro kinase assays and the Luminex technology. Results. Increasing the leptin level in the blood of HS leads to a reduced activation of the oxidative burst by Escherichia coli and phorbol 12-myristate 13-acetate. Activation of the oxidative burst is reduced in the blood of HD patients and the addition of leptin does not lead to further PMNL inhibition. Leptin at a concentration measured in HD patients significantly reduces the chemotaxis of PMNLs from HS but had no effect on PMNLs from ESRD patients before and also after HD treatment with high-flux dialysers. The phosphoinositide 3-kinase/Akt pathway is involved in the inhibitory effects of leptin. Conclusions. In the presence of leptin, PMNLs from HS and HD patients respond differently to stimuli. The lack of response to leptin in PMNLs from HD patients cannot be influenced by HD. PMID:21216885

Cohen, Gerald; Raupachova, Jana; Ilic, Dalibor; Werzowa, Johannes; Horl, Walter H.

2011-01-01

287

Protein-Bound Uremic Toxins Stimulate Crosstalk between Leukocytes and Vessel Wall  

PubMed Central

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

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

2013-01-01

288

Impact of Ambient Air Pollution on the Differential White Blood Cell Count in Patients with Chronic Pulmonary Disease  

PubMed Central

Epidemiologic studies report associations between particulate air pollution and increased mortality from pulmonary diseases.To examine whether the exposure to ambient gaseous and particulate air pollution leads to an alteration of the differential white blood cell count in patients with chronic pulmonary diseases like chronic bronchitis, chronic obstructive pulmonary disease, and asthma. A prospective panel study was conducted in Erfurt, Eastern Germany, with 12 repeated differential white blood cell counts in 38 males with chronic pulmonary diseases. Hourly particulate and gaseous air pollutants and meteorological data were acquired. Mixed models with a random intercept adjusting for trend, meteorology, weekday, and other risk variables were used. In this explorative analysis we found an immediate decrease of polymorphonuclear leukocytes in response to an increase of most gaseous and particulate pollutants. Lymphocytes increased within 24 hours in association with all gaseous pollutants but showed no effect in regard to particulate air pollution. Monocytes showed an increase associated with ultrafine particles, and nitrogen monoxide. The effect had two peaks in time, one 0-23 hours before blood withdrawal and a second one with a time lag of 48-71 hours. The increase of particulate and gaseous air pollution was associated with multiple changes in the differential white blood cell count in patients with chronic pulmonary diseases. PMID:20064088

Bruske, Irene; Hampel, Regina; Socher, Martin M.; Ruckerl, Regina; Schneider, Alexandra; Heinrich, Joachim; Oberdorster, Gunter; Wichmann, H.-Erich; Peters, Annette

2013-01-01

289

Extravasation of leukocytes in comparison to tumor cells  

PubMed Central

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

Strell, Carina; Entschladen, Frank

2008-01-01

290

Selective mobilization of cytotoxic leukocytes by epinephrine.  

PubMed

It is well-known that acute stress, presumably as a first defense against pathogens, enhances PBMC counts by mobilizing these beta2-adrenoceptor positive cells from the marginal pool. Yet, only select leukocyte subsets participate in this phenomenon of adrenergic leukocytosis and underlying mechanisms are obscure. In this study, we analyzed in human blood adhesion molecule and chemokine receptor profiles in 14 leukocyte subsets, and responsiveness of subsets to epinephrine in vivo and in vitro. Five subsets, namely, CCR7(-)CD45RA(+)CD8(+) effector T cells, CD4(-)CD8(-) gamma/delta T cells, CD3(+)CD56(+) NKT-like cells, CD16(+)CD56(dim) cytotoxic NK cells, and CD14(dim)CD16(+) proinflammatory monocytes showed a rapid and transient increase after infusion of epinephrine at physiological concentrations. These cells were characterized by a CD62L(-)CD11a(bright)CX3CR(bright) phenotype, whereby expression of both CD11a and CX3CR1 was strongly correlated with adrenergic leukocytosis in vivo (r = 0.86 and 0.78, p < 0.005). The same subsets showed highest adherence to activated endothelium in vitro, which (except for proinflammatory monocytes) was reversed by epinephrine. We conclude that these five cytotoxic effector leukocyte subsets comprise the marginal pool by a CD11a/CX3CR1-mediated attachment to the endothelium. Epinephrine rapidly attenuates this attachment to allow demargination and release of the cells into the circulation that, because of their cytotoxic effector function, provide immediate protection from invading pathogens. PMID:19949113

Dimitrov, Stoyan; Lange, Tanja; Born, Jan

2010-01-01

291

Antioxidant defenses and lipid peroxidation in human blood plasma.  

PubMed Central

The temporal disappearance in human blood plasma of endogenous antioxidants in relation to the appearance of various classes of lipid hydroperoxides measured by HPLC postcolumn chemiluminescence detection has been investigated under two types of oxidizing conditions. Exposure of plasma to aqueous peroxyl radicals generated at a constant rate leads immediately to oxidation of endogenous ascorbate and sulfhydryl groups, followed by sequential depletion of bilirubin, urate, and alpha-tocopherol. Stimulating polymorphonuclear leukocytes in plasma initiates very rapid oxidation of ascorbate, followed by partial depletion of urate. Once ascorbate is consumed completely, micromolar concentrations of hydroperoxides of plasma phospholipids, triglycerides, and cholesterol esters appear simultaneously, even though sulfhydryl groups, bilirubin, urate, and alpha-tocopherol are still present at high concentrations. Nonesterified fatty acids, the only lipid class in plasma not transported in lipoproteins but bound to albumin, are preserved from peroxidative damage even after complete oxidation of ascorbate, most likely due to site-specific antioxidant protection by albumin-bound bilirubin and possibly by albumin itself. Thus, in plasma ascorbate and, in a site-specific manner, bilirubin appear to be much more effective in protecting lipids from peroxidative damage by aqueous oxidants than all the other endogenous antioxidants. Hydroperoxides of linoleic acid, phosphatidylcholine, and cholesterol added to plasma in the absence of added reducing substrates are degraded, in contrast to hydroperoxides of trilinolein and cholesterol linoleate. These findings indicate the presence of a selective peroxidase activity operative under physiological conditions. Our data suggest that in states of leukocyte activation and other types of acute or chronic oxidative stress such a simple regimen as controlled ascorbate supplementation could prove helpful in preventing formation of lipid hydroperoxides, some of which cannot be detoxified by endogenous plasma activities and thus might cause damage to critical targets. PMID:3200852

Frei, B; Stocker, R; Ames, B N

1988-01-01

292

Primary role for adherent leukocytes in sickle cell vascular occlusion: A new paradigm  

PubMed Central

Vascular occlusion is the major cause of morbidity and mortality in sickle cell disease but its mechanisms are poorly understood. We demonstrate by using intravital microscopy in mice expressing human sickle hemoglobin (SS) that SS red blood cells (RBCs) bind to adherent leukocytes in inflamed venules, producing vasoocclusion of cremasteric venules. SS mice deficient in P- and E-selectins, which display defective leukocyte recruitment to the vessel wall, are protected from vasoocclusion. These data uncover a previously unsuspected paradigm for the pathogenesis of sickle cell vasoocclusion in which adherent leukocytes play a direct role and suggest that drugs targeting SS RBC–leukocyte or leukocyte–endothelial interactions may prevent or treat the vascular complications of this debilitating disease. PMID:11880644

Turhan, Aslihan; Weiss, Linnea A.; Mohandas, Narla; Coller, Barry S.; Frenette, Paul S.

2002-01-01

293

Altered Mitochondrial Function and Oxidative Stress in Leukocytes of Anorexia Nervosa Patients  

PubMed Central

Context Anorexia nervosa is a common illness among adolescents and is characterised by oxidative stress. Objective The effects of anorexia on mitochondrial function and redox state in leukocytes from anorexic subjects were evaluated. Design and setting A multi-centre, cross-sectional case-control study was performed. Patients Our study population consisted of 20 anorexic patients and 20 age-matched controls, all of which were Caucasian women. Main outcome measures Anthropometric and metabolic parameters were evaluated in the study population. To assess whether anorexia nervosa affects mitochondrial function and redox state in leukocytes of anorexic patients, we measured mitochondrial oxygen consumption, membrane potential, reactive oxygen species production, glutathione levels, mitochondrial mass, and complex I and III activity in polymorphonuclear cells. Results Mitochondrial function was impaired in the leukocytes of the anorexic patients. This was evident in a decrease in mitochondrial O2 consumption (P<0.05), mitochondrial membrane potential (P<0.01) and GSH levels (P<0.05), and an increase in ROS production (P<0.05) with respect to control subjects. Furthermore, a reduction of mitochondrial mass was detected in leukocytes of the anorexic patients (P<0.05), while the activity of mitochondrial complex I (P<0.001), but not that of complex III, was found to be inhibited in the same population. Conclusions Oxidative stress is produced in the leukocytes of anorexic patients and is closely related to mitochondrial dysfunction. Our results lead us to propose that the oxidative stress that occurs in anorexia takes place at mitochondrial complex I. Future research concerning mitochondrial dysfunction and oxidative stress should aim to determine the physiological mechanism involved in this effect and the physiological impact of anorexia. PMID:25254642

Victor, Victor M.; Rovira-Llopis, Susana; Saiz-Alarcon, Vanessa; Sanguesa, Maria C.; Rojo-Bofill, Luis; Banuls, Celia; Falcon, Rosa; Castello, Raquel; Rojo, Luis; Rocha, Milagros; Hernandez-Mijares, Antonio

2014-01-01

294

Pretreatment with polynitroxyl albumin (PNA) inhibits ischemia-reperfusion induced leukocyte-endothelial cell adhesion.  

PubMed

Recently published evidence indicates that polynitroxylated albumin (PNA) protects tissues against ischemia/reperfusion (I/R) injury, possibly by enhancing tissue redox activity. The objective of this study was to determine if PNA treatment alters the leukocyte-endothelial cell adhesion that is normally elicited by I/R. PNA, human serum albumin (HSA) or saline were administered (i.v.) 5 min before reperfusion. Venular diameter, red blood cell velocity, wall shear rate, systemic hematocrit, systemic arterial pressure, as well as the number of adherent and emigrated leukocytes were monitored in rat mesenteric venules before and after 20 min of ischemia and 30 min of reperfusion. In saline-treated rats, I/R elicited a 5.3-fold increase in leukocyte adhesion and a 1.8-fold increase in leukocyte emigration. HSA-treated animals exhibited 4.0 and 2.3-fold increases in leukocyte adherence and emigration, respectively. In PNA-treated rats, the number of adherent leukocytes increased only 2.1-fold increase in adherent leukocytes, while leukocyte emigration was completely inhibited. The PNA-induced attenuation of leukocyte adherence/emigration could not be attributed to alterations in systemic or local hemodynamics (red blood cell velocity or wall shear rate). PNA was also shown to be a potent inhibitor of xanthine-xanthine oxidase mediated adhesion of human neutrophils to cultured human endothelial cells. These findings indicate that PNA may protect tissues against I/R injury by attenuating leukocyte-endothelial cell adhesion. PMID:9667490

Russell, J; Okayama, N; Alexander, J S; Granger, D N; Hsia, C J

1998-07-15

295

White blood cell count - series (image)  

MedlinePLUS

The White Blood Cell (WBC) Count measures two components: the total number of WBC's (leukocytes), and the differential count. ... and basophils) and non-granulocytes (lymphocytes and monocytes). White blood cells are a major component of the ...

296

A Role for Lactate Dehydrogenases in the Survival of Neisseria gonorrhoeae in Human Polymorphonuclear Leukocytes and Cervical Epithelial Cells.  

PubMed

Lactate is an abundant metabolite, produced by host tissues and commensal organisms, and it represents an important potential carbon source for bacterial pathogens. In the case of Neisseria spp., the importance of the lactate permease in colonization of the host has been demonstrated, but there have been few studies of lactate metabolism in pathogenic Neisseria in the postgenomic era. We describe herein the characterization of genome-annotated, respiratory, and substrate-level lactate dehydrogenases (LDHs) from the obligate human pathogen Neisseria gonorrhoeae. Biochemical assays using N. gonorrhoeae 1291 wild type and isogenic mutant strains showed that cytoplasmic LdhA (NAD(+)-dependent D-lactate dehydrogenase) and the membrane-bound respiratory enzymes, LdhD (D-lactate dehydrogenase) and LldD (L-lactate dehydrogenase) are correctly annotated. Mutants lacking LdhA and LdhD showed greatly reduced survival in neutrophils compared with wild type cells, highlighting the importance of D-lactate metabolism in gonococcal survival. Furthermore, an assay of host colonization using the well-established human primary cervical epithelial cell model revealed that the two respiratory enzymes make a significant contribution to colonization of and survival within the microaerobic environment of the host. Taken together, these data suggest that host-derived lactate is critical for the growth and survival of N. gonorrhoeae in human cells. PMID:24737798

Atack, John M; Ibranovic, Ines; Ong, Cheryl-Lynn Y; Djoko, Karrera Y; Chen, Nathan H; Vanden Hoven, Rachel; Jennings, Michael P; Edwards, Jennifer L; McEwan, Alastair G

2014-10-15

297

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

E-print Network

there is little evidence in the literature that clinical abnormalities exist in PHA dogs, that there are subtle abberations in phagocytic capabilities that would manifest themselves under extreme challenge or stress. Phagocytosis and killing of microbes... simultaneously to evaluate uptake and killing of microbes as two separate events in the continuum of the phagocytic process. LITERATURE REVIEW Pelger-Huet Anomaly (PHA) is the most common congenital anomaly involving the human neutrophil nucleus...

Browder, Elizabeth Jarratt

2012-06-07

298

Regulation of Oscillations in Filamentous Actin Content in Polymorphonuclear Leukocytes Stimulated with Leukotriene B 4 and Platelet-Activating Factor  

Microsoft Academic Search

Stimulation of neutrophils with LTB4 or PAF results in the production of a rapidly oscillating actin polymerization\\/depolymerization response. Treatment of neutrophils with inhibitors of PKC prior to stimulation with ligand resulted in a masking of the F-actin oscillations. Because myosin has been shown to be a substrate for neutrophil PKC, this protein was investigated as a potential downstream mediator of

Ramesh Rengan; Geneva M. Omann

1999-01-01

299

The activation of gold complexes by cyanide produced by polymorphonuclear leukocytes. III. The formation of aurocyanide by myeloperoxidase.  

PubMed

There is considerable evidence that the anti-rheumatic gold complexes are activated by their conversion to aurocyanide. In order to understand the mechanism of production of aurocyanide, we investigated the involvement of myeloperoxidase in the reaction. This haem enzyme of neutrophils and monocytes uses hydrogen peroxide to oxidise chloride and thiocyanate to hypochlorous acid and hypothiocyanite, respectively. When aurothiomalate (10 microM) was incubated with thiocyanate (200 microM), hydrogen peroxide (100 microM) and myeloperoxidase (20 nM), it was transformed to a product that was spectrally identical to authentic aurocyanide. Aurothiomalate was quantitatively converted to aurocyanide in about 10 min at pH 6.0 and in 40 min at pH 7.4. Aurocyanide formation occurred after myeloperoxidase had used all the hydrogen peroxide available to produce hypothiocyanite. Thus, the cyanide must have formed from the slow decomposition of hypothiocyanite. The rate of aurocyanide production was increased in the presence of 100 mM chloride, which indicates that hypochlorous acid accelerates the formation of cyanide. Hypochlorous acid (100 to 400 microM) reacted non-enzymatically with thiocyanate (200 microM) and aurothiomalate (10 microM) to produce aurocyanide. Thus, aurocyanide is produced by two processes, involving both the formation of hypothiocyanite and hypochlorous acid. Aurocyanide is an effective inhibitor of the respiratory burst of neutrophils and monocytes and the proliferation of lymphocytes. Therefore, aurothiomalate may attenuate inflammation by acting as a pro-drug which is reliant on neutrophils and monocytes to produce hypothiocyanite. When the hypothiocyanite decays to hydrogen cyanide, the pro-drug is converted to aurocyanide which then suppresses further oxidant production by these inflammatory cells. PMID:9744567

Graham, G G; Kettle, A J

1998-08-01

300

Initial blood storage experiment  

NASA Technical Reports Server (NTRS)

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.

Surgenor, Douglas MACN.

1988-01-01

301

Modulation of in vivo immune response by selective depletion of neutrophils using a monoclonal antibody, RP-3. II. Inhibition by RP-3 treatment of mononuclear leukocyte recruitment in delayed-type hypersensitivity to sheep red blood cells in rats.  

PubMed

We demonstrated in our previous paper that treatment of rats with RP-3, a mAb that depletes in vivo neutrophils selectively, resulted in inhibition of both the priming and effector phases of delayed type hypersensitivity (DTH) to SRBC. In order to clarify the mechanisms of this phenomenon, we examined the effect of RP-3 treatment on mononuclear leukocyte (MNL) recruitment in DTH. When rats had been treated with RP-3 at the time of Ag priming, MNL migration, which accompanies DTH, was inhibited. The prior infiltration of neutrophils was also partially required for MNL recruitment because migration was inhibited when the immune rats were treated with RP-3 at the time of DTH elicitation. PMID:8473730

Kudo, C; Yamashita, T; Terashita, M; Sendo, F

1993-05-01

302

Photodynamic therapy induced leukocyte adherence in normal subcutaneous, but not in tumor microvessels  

NASA Astrophysics Data System (ADS)

Leukocyte mediated tissue damage is proposed to contribute to the tumoricidal effect of photodynamic therapy (PDT). To elucidate this hypothesis leukocyte-endothelium interaction and red blood cell velocity in the microvasculature of an amelanotic melanoma and normal tissue in a skin fold chamber preparation were investigated before and after PDT (5 mg/kg PhotofrinR; 630 nm; 10 or 100 J/cm2). One hour after PDT (10 and 100 J/cm2) the number of adhering leukocytes increased in subcutaneous but not in tumor microvessels. At this time red blood cell velocity remained unaltered in microvessels of subcutaneous tissue but decreased (10 J/cm2) and even ceased (100 J/cm2) in tumor microvessels. These results indicate that tumor destruction induced by PDT is not mediated by leukocyte-endothelium interaction.

Dellian, Marc; Abels, Christoph; Kuhnle, Gerhard E.; Goetz, Alwin E.

1995-03-01

303

Photodynamic therapy induced leukocyte adherence in normal subcutaneous, but not in tumor microvessels  

NASA Astrophysics Data System (ADS)

Leukocyte mediated tissue damage is proposed to contribute to the tumoricidal effect of photodynamic therapy (PDT). To elucidate this hypothesis leukocyte-endothelium interaction and red blood cell velocity in the microvasculature of an amelanotic melanoma and normal tissue in a skin fold chamber preparation were investigated before and after PDT (5 mg/kg PhotofrinR; 630 nm; 10 or 100 J/cm2). One hour after PDT (10 and 100 J/cm2) the number of adhering leukocytes increased in subcutaneous but not in tumor microvessels. At this time red blood cell velocity remained unaltered in microvessels of subcutaneous tissue but decreased (10 J/cm2) and even ceased (100 J/cm2) in tumor microvessels. These results indicate that tumor destruction induced by PDT is not mediated by leukocyte-endothelium interaction.

Dellian, Marc; Abels, Christoph; Kuhnle, Gerhard E.; Goetz, Alwin E.

1994-10-01

304

Leukocyte-endothelial cell adhesive interactions: role of xanthine oxidase-derived oxidants.  

PubMed

The objective of this study was to determine whether agents that either scavenge or inhibit the production of oxygen radicals can alter the adhesive interactions between leukocytes and venular endothelium elicited by ischemia-reperfusion. Cat mesenteric and intestinal blood flows were reduced to 20% of baseline for 1 hr, followed by 1 hr of reperfusion. Sixty minutes after reperfusion, red blood cell velocity (Vr), leukocyte rolling velocity (Vw), and the number of adherent leukocytes were measured in mesenteric venules. Then, either manganese-superoxide dismutase (Mn-SOD), catalase, desferrioxamine, or oxypurinol was administered intravascularly. Ten minutes later, repeat measurements were obtained and compared with pretreatment values. Catalase, Mn-SOD, and oxypurinol significantly attenuated neutrophil adherence while neither inactivated-catalase nor desferrioxamine altered the reperfusion-induced leukocyte adhesion. The ratio of Vw to erythrocyte velocity, an index of the fracture stress between rolling leukocytes and venular endothelium, was not altered by any of the agents studied. These results and data in the literature indicate that many of the agents that are commonly used to either scavenge or inhibit the production of oxygen radicals in postischemic tissues exert a significant inhibitory influence on leukocyte adhesion to microvascular endothelium in vivo. Our results are also consistent with the view that xanthine oxidase-derived oxidants contribute to the leukocyte-endothelial cell adhesive interactions associated with reperfusion of ischemic tissues. PMID:1748842

Suzuki, M; Grisham, M B; Granger, D N

1991-11-01

305

A multiscale SPH particle model of the near-wall dynamics of leukocytes in flow.  

PubMed

A novel multiscale Lagrangian particle solver based on SPH is developed with the intended application of leukocyte transport in large arteries. In such arteries, the transport of leukocytes and red blood cells can be divided into two distinct regions: the bulk flow and the near-wall region. In the bulk flow, the transport can be modeled on a continuum basis as the transport of passive scalar concentrations. Whereas in the near-wall region, specific particle tracking of the leukocytes is required and lubrication forces need to be separately taken into account. Because of large separation of spatio-temporal scales involved in the problem, simulations of red blood cells and leukocytes are handled separately. In order to take the exchange of leukocytes between the bulk fluid and the near-wall region into account, solutions are communicated through coupling of conserved quantities at the interface between these regions. Because the particle tracking is limited to those leukocytes lying in the near-wall region only, our approach brings considerable speedup to the simulation of leukocyte circulation in a test geometry of a backward-facing step, which encompasses many flow features observed in vivo. PMID:24009138

Gholami, Babak; Comerford, Andrew; Ellero, Marco

2014-01-01

306

Leukocytes as mediators of pain and analgesia  

Microsoft Academic Search

In inflammation, resident cells and infiltrating leukocytes produce proalgesic mediators. Although these mediators induce\\u000a pain, the role of specific cell populations is still controversial. In addition, resident cells and leukocytes also generate\\u000a analgesic mediators that counteract inflammatory pain, including anti-inflammatory cytokines, endocannabinoids, and opioid\\u000a peptides. Chemokines and adhesion molecules orchestrate the migration of opioid peptide-containing leukocytes to inflamed\\u000a tissue. Leukocytes

Heike L. Rittner; Alexander Brack

2007-01-01

307

CD47 deficiency does not impede polymorphonuclear neutrophil transmigration but attenuates granulopoiesis at the postacute stage of colitis.  

PubMed

Previous studies have suggested that CD47, an essential cell-surface protein, plays an important role in polymorphonuclear neutrophil (PMN) transmigration across tissue cells and extracellular matrix. In the current study, the role of CD47 in PMN transmigration and infiltration into tissues was further evaluated by investigating the function of CD47(-/-) PMN and inflammatory conditions induced in CD47(-/-) mice. Using in vitro time-course assays, we found that CD47(-/-) PMN exhibited no impediment, but slightly enhanced response to and transmigration toward, the chemoattractant fMLF. In vivo analysis in CD47(-/-) mice by inducing acute peritonitis and aggressive colitis observed consistent results, indicating that both PMN and monocytes effectively infiltrated inflammatory sites despite the absence of CD47 on these leukocytes or the surrounding tissue cells. Although PMN transmigration was not delayed in CD47(-/-) mice, fewer PMN were found in the intestine at the postacute/chronic stage of chronic colitis induced with sustained low-dose dextran sulfate sodium. Further analysis suggested that the paucity of PMN accumulation was attributable to attenuated granulopoiesis secondary to assessed lower levels of IL-17. Administration of exogenous IL-17A markedly increased PMN availability and rapidly rendered severe colitis in CD47(-/-) mice under dextran sulfate sodium treatment. PMID:23203922

Bian, Zhen; Guo, Yalan; Luo, Youqun; Tremblay, Alexandra; Zhang, Xiugen; Dharma, Sanam; Mishra, Aarti; Liu, Yuan

2013-01-01

308

Rapid in vitro biocompatibility assay of endovascular stents by flow cytometry using platelet activation and platelet-leukocyte aggregation.  

PubMed

Clinical studies suggest that stent design and surface texture are responsible for differences in biocompatibility of metallic endovascular stents. A simple in vitro experimental setup was established to test stent-induced degree of platelet and leukocyte activation and platelet-leukocyte aggregation by flow cytometry. Heparin-coated tantalum stents and gold-coated and uncoated stainless steel stents were tested. Stents were implanted into silicone tubes and exposed to blood from healthy volunteers. Platelet and leukocyte activation and percentage of leukocyte-platelet aggregates were determined in a whole-blood assay by subsequent staining for activation-associated antigens (CD41a, CD42b, CD62p, and fibrinogen binding) and leukocyte antigens (CD14 and CD45) and flow cytometric analysis. Blood taken directly after venous puncture or exposed to the silicone tube alone was used as negative controls. Positive control was in vitro stimulation with thrombin receptor activating peptide (TRAP-6). Low degree of platelet activation and significant increase in monocyte- and neutrophil-platelet aggregation were observed in blood exposed to stents (P < 0.05). In addition, leukocyte activation was induced as measured by increased CD45 and CD14 expression. Heparin coated stents continuously induced less platelet activation and leukocyte-platelet aggregation than uncoated stainless steel stents of the same length and shorter stents of the same structure. Stent surface coating and texture plays a role in platelet and leukocyte activation and leukocyte-platelet aggregation. Using this simple in vitro assay and whole blood and flow cytometry, it seems possible to differentiate stents by their potency to activate platelets and/or leukocytes. This assay could be applied for improving the biocompatibility of coronary stents. PMID:10088974

Tárnok, A; Mahnke, A; Müller, M; Zotz, R J

1999-02-15

309

Chemiluminescence induced by phagocytosis of Escherichia coli by polymorphonuclear leucocytes.  

PubMed

Chemiluminescence emitted by phagocytosing human polymorphonuclear leucocytes stimulated by Escherichia coli was measured using a liquid scintillation counter equipped with a multichannel analyser. In the presence of the amplifying agent luminol, light emission can be divided into two channels, one of which ('high energy') appears to correlate directly with phagocytic activity of the PMNL, and the other ('low energy') with the background luminol dioxygenation by the cells. Measuring in the 'high energy' window also eliminates the normal 'out of coincidence' background. The method is applicable to measuring opsonizing capacity of different sera, and responds to PMNL number, age, composition of assay medium and the integrity of the stimulating bacteria. Other bacterial strains produce a similar response, as does the artificial stimulator zymosan. Low temperature and anaerobiosis, which inhibit phagocytic killing, also suppress light emission. PMID:6389762

Fazeli, A; Richards, L

1984-09-01

310

Sesquiterpene lactone from Wunderlichia crulsiana inhibits the respiratory burst of leukocytes triggered by distinct biochemical pathways.  

PubMed

The sesquiterpene lactone tubiferin was chemically purified from the brazilian native plant Wunderlichia crulsiana and identified by NMR and GC/MS data. Its ability to inhibit the respiratory burst of peritoneal inflammatory polymorphonuclear leukocytes (PMN) stimulated upon addition of phorbol miristate acetate (PMA), opsonized zymosan (OZ), and N-formyl-methionyl-leucyl-phenylalanine (fMLP) was evaluated. The tubiferin inhibition was more pronounced when PMN were stimulated through the protein kinase C pathway (PMA) compared to the alternative complement pathway (OZ). The inhibition when PMN were triggered by a chemoattractant stimulus (fMLP) was similar to that achieved with OZ-stimulated phagocytes. Tubiferin showed dose-dependent effects on the PMN respiratory burst triggered by the three different substances, and also decreased substantially the carrageenan-induced mice paw edema. PMID:12927587

Nuñez, Cecilia Veronica; Zacheu, Fabíola Maria; Pinto, Ernani; Roque, Nídia Franca; Colepicolo, Pio; Brigagão, Maísa Ribeiro Pereira Lima

2003-09-12

311

Leukocyte driven-decidual angiogenesis in early pregnancy  

PubMed Central

Successful pregnancy and long-term, post-natal maternal and offspring cardiac, vascular and metabolic health require key maternal cardiovascular adaptations over gestation. Within the pregnant decidualizing uterus, coordinated vascular, immunological and stromal cell changes occur. Considerable attention has been given to the roles of uterine natural killer (uNK) cells in initiating decidual spiral arterial remodeling, a process normally completed by mid-gestation in mice and in humans. However, leukocyte roles in much earlier, region specific, decidual vascular remodeling are now being defined. Interest in immune cell-promoted vascular remodeling is driven by vascular aberrations that are reported in human gestational complications such as infertility, recurrent spontaneous abortion, preeclampsia (PE) and fetal growth restriction. Appropriate maternal cardiovascular responses during pregnancy protect mothers and their children from later cardiovascular disease risk elevation. One of the earliest uterine responses to pregnancy in species with hemochorial placentation is stromal cell decidualization, which creates unique niches for angiogenesis and leukocyte recruitment. In early decidua basalis, the aspect of the implantation site that will cradle the developing placenta and provide the major blood vessels to support mature placental functions, leukocytes are greatly enriched and display specialized properties. UNK cells, the most abundant leukocyte subset in early decidua basalis, have angiogenic abilities and are essential for normal early decidual angiogenesis. The regulation of uNK cells and their roles in determining maternal and progeny cardiovascular health over pregnancy and postpartum are discussed. PMID:25066422

Lima, Patricia DA; Zhang, Jianhong; Dunk, Caroline; Lye, Stephen J; Anne Croy, B

2014-01-01

312

Leukocyte margination at arteriole shear rate  

PubMed Central

Abstract We numerically investigated margination of leukocytes at arteriole shear rate in straight circular channels with diameters ranging from 10 to 22 ?m. Our results demonstrated that passing motion of RBCs effectively induces leukocyte margination not only in small channels but also in large channels. A longer time is needed for margination to occur in a larger channel, but once a leukocyte has marginated, passing motion of RBCs occurs continuously independent of the channel diameter, and leukocyte margination is sustained for a long duration. We also show that leukocytes rarely approach the wall surface to within a microvillus length at arteriole shear rate. PMID:24907300

Takeishi, Naoki; Imai, Yohsuke; Nakaaki, Keita; Yamaguchi, Takami; Ishikawa, Takuji

2014-01-01

313

Gene Expression Profile of Endotoxin-stimulated Leukocytes of the Term New Born: Control of Cytokine Gene Expression by Interleukin-10  

PubMed Central

Introduction Increasing evidence now supports the association between the fetal inflammatory response syndrome (FIRS) with the pathogenesis of preterm labor, intraventricular hemorrhage and bronchopulmonary dysplasia. Polymorphonuclear leukocyte (PMNs) and mononuclear cell (MONOs) infiltration of the placenta is associated with these disorders. The aim of this study was to reveal cell-specific differences in gene expression and cytokine release in response to endotoxin that would elucidate inflammatory control mechanisms in the newly born. Methods PMNs and MONOs were separately isolated from the same cord blood sample. A genome-wide microarray screened for gene expression and related pathways at 4 h of LPS stimulation (n?=?5). RT-qPCR and ELISA were performed for selected cytokines at 4 h and 18 h of LPS stimulation. Results Compared to PMNs, MONOs had a greater diversity and more robust gene expression that included pro-inflammatory (PI) cytokines, chemokines and growth factors at 4 h. Only MONOs had genes changing expression (all up regulated including interleukin-10) that were clustered in the JAK/STAT pathway. Pre-incubation with IL-10 antibody, for LPS-stimulated MONOs, led to up regulated PI and IL-10 gene expression and release of PI cytokines after 4 h. Discussion The present study suggests a dominant role of MONO gene expression in control of the fetal inflammatory response syndrome at 4 hrs of LPS stimulation. LPS-stimulated MONOs but not PMNs of the newborn have the ability to inhibit PI cytokine gene expression by latent IL-10 release. PMID:23326478

Davidson, Dennis; Zaytseva, Alla; Miskolci, Veronika; Castro-Alcaraz, Susana; Vancurova, Ivana; Patel, Hardik

2013-01-01

314

Fluorescently Activated Cell Sorting Followed by Microarray Profiling of Helper T Cell Subtypes from Human Peripheral Blood  

PubMed Central

Background Peripheral blood samples have been subjected to comprehensive gene expression profiling to identify biomarkers for a wide range of diseases. However, blood samples include red blood cells, white blood cells, and platelets. White blood cells comprise polymorphonuclear leukocytes, monocytes, and various types of lymphocytes. Blood is not distinguishable, irrespective of whether the expression profiles reflect alterations in (a) gene expression patterns in each cell type or (b) the proportion of cell types in blood. CD4+ Th cells are classified into two functionally distinct subclasses, namely Th1 and Th2 cells, on the basis of the unique characteristics of their secreted cytokines and their roles in the immune system. Th1 and Th2 cells play an important role not only in the pathogenesis of human inflammatory, allergic, and autoimmune diseases, but also in diseases that are not considered to be immune or inflammatory disorders. However, analyses of minor cellular components such as CD4+ cell subpopulations have not been performed, partly because of the limited number of these cells in collected samples. Methodology/Principal Findings We describe fluorescently activated cell sorting followed by microarray (FACS–array) technology as a useful experimental strategy for characterizing the expression profiles of specific immune cells in the circulation. We performed reproducible gene expression profiling of Th1 and Th2, respectively. Our data suggest that this procedure provides reliable information on the gene expression profiles of certain small immune cell populations. Moreover, our data suggest that GZMK, GZMH, EOMES, IGFBP3, and STOM may be novel markers for distinguishing Th1 cells from Th2 cells, whereas IL17RB and CNTNAP1 can be Th2-specific markers. Conclusions/Significance Our approach may help in identifying aberrations and novel therapeutic or diagnostic targets for diseases that affect Th1 or Th2 responses and elucidating the involvement of a subpopulation of immune cells in some diseases. PMID:25379667

Ono, Chiaki; Yu, Zhiqian; Kasahara, Yoshiyuki; Kikuchi, Yoshie; Ishii, Naoto; Tomita, Hiroaki

2014-01-01

315

Leukotriene B4 mediates shear rate-dependent leukocyte adhesion in mesenteric venules.  

PubMed

Previous studies have demonstrated that low shear rates promote leukocyte adherence to microvascular endothelium in postcapillary venules. The objective of this study was to determine whether an accumulation of inflammatory mediators such as platelet activating factor and leukotriene B4 is responsible for shear rate-dependent leukocyte-endothelial cell adhesion. Postcapillary venules (25-39 microns in diameter) in cat mesentery were studied by intravital microscopy. Venular wall shear rate was varied over a wide range by graded occlusion of the mesenteric artery. Red blood cell velocity, vessel diameter, leukocyte rolling velocity, and the numbers of rolling and adherent leukocytes were measured at each shear rate. In one series of experiments, shear rate-dependent leukocyte adherence was monitored at different superfusion rates (1.0 and 2.5 ml/min). At the lower superfusion rate, the number of adherent leukocytes was significantly higher at any given shear rate when compared with results obtained at the higher superfusion rate. This suggests that reduced washout of inflammatory mediators contributes to shear rate-dependent leukocyte adhesion. Pretreatment with different platelet activating factor receptor antagonists (WEB 2086 or WEB 2170) had no effect on the number of adherent leukocytes normally observed at lower shear rates, suggesting that platelet activating factor does not play a major role in this process. However, shear rate-dependent leukocyte adhesion was largely prevented by pretreatment with either a leukotriene B4 receptor antagonist (SC-41930) or a leukotriene synthesis inhibitor (L663,536). The results of this study indicate that a reduced washout of leukotriene B4 is responsible for the enhanced leukocyte adherence that occurs at low venular wall shear rates. PMID:1355410

Bienvenu, K; Russell, J; Granger, D N

1992-10-01

316

A comparative study of bovine blood and milk neutrophil functions with luminol-dependent chemiluminescence.  

PubMed

In this study, a technique was developed for the chemiluminescence (CL) measurement of bovine milk polymorphonuclear leukocytes (PMN). In the first study, the effects of cell number and the concentration of phorbol-12-myristate-13-acetate (PMA), luminol, latex bead particles, dimethyl sulphoxide (DMSO) and gelatin on the luminol-dependent cellular CL (LDCL) response were assessed with healthy cows in different stages of lactation. In the second study, the LDCL and in vitro bactericidal activity of blood and milk PMN towards Staphylococcus aureus was investigated. In general, the CL activity of blood PMN was consistently higher than that of milk PMN. We found that (a) the optimal cell density in blood and milk cells for maximal LDCL response ranged from 1.5 x 10(6) to 5 x 10(6) cells/mL; (b) the optimal concentrations of PMA, latex beads and luminol for maximal LDCL response were 100-200 ng/ml, 500 particles/PMN and 0.1 mmol/L, respectively. Concentrations of DMSO of 0.5-1% (v/v) did not significantly affect the maximal CL response of PMN. Gelatin concentrations of 0.1 -0.5 mg/ml had no effect on the LDCL of PMN. In addition, the LDCL of PMN was significantly correlated with bactericidal activity towards S. aureus (r = 0.78, p < 0.001 for blood PMN and r = 0.66, p < 0.01 for milk PMN). Under the optimal experimental conditions for measurement of CL produced by bovine blood and milk PMN defined in this study, LDCL assay is an accurate and reproducible technique for the rapid quantification of PMN bactericidal activity in physiological and pathological conditions of high-yielding dairy cows. PMID:11754137

Mehrzad, J; Dosogne, H; Vangroenweghe, F; Burvenich, C

2001-01-01

317

Concanavalin A as a probe for studying the mechanism of metabolic stimulation of leukocytes.  

PubMed

The disruption of the molecular organization of the plasma membrane of leukocytes by phagocytosable particles, or by agents such as surfactants, antibodies, phospholipase C, fatty acids and chemotactic factors, leads to a stimulation of the phagocyte oxidative metabolism. Concanavalin A (Con A) has been used as a tool to study the mechanism of this metabolic regulation. The binding of Con A to the surface of polymorphonuclear leukocytes (PMNL) or macrophages produces a rapid enhancement of oxygen uptake and glucose oxidation through the hexose monophosphate pathway (HMP). This is explained by an activation of the granular NADPH oxidase, the key enzyme in the metabolic stimulation. The effect of Con A is not due to endocytosed lectin, since Con A covalently coupled to large sepharose beads still acts as stimulant. The metabolic changes caused by Con A are reversible. If, after the onset of stimulation, sugars with high affinity for Con A are added to the leukocyte suspension, the activity of granular NADPH oxidase and the rate of respiration and glucose oxidation return to their resting values. The metabolic burst, while partially supressed by treatment of PMNL with iodoacetate, sodium flouride and cytochalasin B, is slightly increased by colchicine. Con A induces a selective release of granular enzymes (beta-glucuronidase, peroxidase, alkaline phosphatase) from PMNL, whereas no leakage of cytoplasmic enzymes is observed. The enzyme release is inhibited by iodoacetate and by drugs known to increase cell levels of cyclic AMP. Based on a current view of the mode of interaction between Con A and cell surfaces, a model of the metabolic disruption of leukocytes is presented. PMID:168745

Romeo, D; Zabucchi, G; Jug, M; Miani, N; Soranzo, M R

1975-01-01

318

Platelet-activating factor promotes shear rate-dependent leukocyte adhesion in postcapillary venules.  

PubMed

Reductions in shear rate are generally associated with a recruitment of rolling and adherent leukocytes within the microcirculation. In this study, we determined if platelet-activating factor (PAF) affects shear rate-dependent leukocytes adhesion in cat mesenteric venules and whether the adhesion glycoproteins CD11/CD18 and ICAM-1 contribute to this process. Shear rate was varied in postcapillary venules (25-35 microns) by graded occlusion of the superior mesenteric artery. Vessel diameter, red blood cell velocity, leukocyte rolling velocity, and the number of rolling and adherent leukocytes were measured at each shear rate either in the presence or absence of PAF superfusion. PAF superfusion resulted in an increased number of adherent, but not rolling, leukocytes at basal and reduced venular shear rates. A monoclonal antibody (MAb) directed against CD11/CD18 prevented the PAF-induced recruitment of adherent leukocytes, while an ICAM-1 specific MAb had no effect. These results indicate that PAF increases shear rate-dependent leukocyte adhesion, presumably through activation and/or increased surface expression of CD11/CD18 on granulocytes. PMID:7903566

Bienvenu, K; Russell, J; Granger, D N

1993-10-01

319

Sialylated, fucosylated ligands for L-selectin expressed on leukocytes mediate tethering and rolling adhesions in physiologic flow conditions  

PubMed Central

Interaction of leukocytes in flow with adherent leukocytes may contribute to their accumulation at sites of inflammation. Using L- selectin immobilized in a flow chamber, a model system that mimics presentation of L-selectin by adherent leukocytes, we characterize ligands for L-selectin on leukocytes and show that they mediate tethering and rolling in shear flow. We demonstrate the presence of L- selectin ligands on granulocytes, monocytes, and myeloid and lymphoid cell lines, and not on peripheral blood T lymphocytes. These ligands are calcium dependent, sensitive to protease and neuraminidase, and structurally distinct from previously described ligands for L-selectin on high endothelial venules (HEV). Differential sensitivity to O-sialo- glycoprotease provides evidence for ligand activity on both mucin-like and nonmucin-like structures. Transfection with fucosyltransferase induces expression of functional L-selectin ligands on both a lymphoid cell line and a nonhematopoietic cell line. L-selectin presented on adherent cells is also capable of supporting tethering and rolling interactions in physiologic shear flow. L-selectin ligands on leukocytes may be important in promoting leukocyte-leukocyte and subsequent leukocyte endothelial interactions in vivo, thereby enhancing leukocyte localization at sites of inflammation. PMID:8909555

1996-01-01

320

Channel catfish (Ictalurus punctatus) leukocytes express estrogen receptor isoforms ER? and ER?2 and are functionally modulated by estrogens  

USGS Publications Warehouse

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.

Iwanowicz, Luke R.; Stafford, James L.; Patiño, Reynaldo; Bengten, Eva; Miller, Norman W.; Blazer, Vicki S.

2014-01-01

321

Channel catfish (Ictalurus punctatus) leukocytes express estrogen receptor isoforms ER? and ER?2 and are functionally modulated by estrogens.  

PubMed

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

Iwanowicz, Luke R; Stafford, James L; Patiño, Reynaldo; Bengten, Eva; Miller, Norman W; Blazer, Vicki S

2014-09-01

322

5-Methyl-4H-3,1-benzoxazin-4-one derivatives: specific inhibitors of human leukocyte elastase.  

PubMed

Inhibitors of human leukocyte elastase (HLE) may exert potent therapeutic effects on pulmonary emphysema, adult respiratory distress syndrome and other diseases involving tissue degradation. 7-(4-Chlorophenylsulfonyl-L-glutanyl)amino-5-methyl-2-isopro pylamino-4H-3,1- benzoxazin-4-one (TEI-5624) and 7-(4-chlorophenylsulfonyl-L-lysyl)amino-5-methyl-2- isopropylamino-4H-3,1-benzoxazin-4-one (TEI-6344), two derivatives of 5-methyl-4H-3,1-benzoxazin-4-one, showed strong and highly specific inhibition of human sputum elastase (HSE), which is equivalent to HLE, with Ki values of 6.91 and 16.3 nM, respectively. The selectivity of TEI-5624 for HSE vs. several proteinases ranged from 300-fold to 45,000-fold in favor of HSE. TEI-5624 and TEI-6344 also efficiently prevented degradation of insoluble elastin by stimulated polymorphonuclear leukocytes. The elastase inhibitory capacity of these compounds was not affected by treatment with stimulated polymorphonuclear leukocytes or Pseudomonas aeruginosa-origin elastase. When administered intratracheally to hamsters. TEI-5624 and TEI-6344 were eliminated from the lung with half-times of 85 and 240 min, respectively. In acute injury induced by intratracheal administration of HSE in hamsters, these compounds significantly suppressed pulmonary hemorrhage when administered intratracheally (1 mg/kg) either 30 or 240 min before challenge with HSE (1 mg/kg). HSE-induced emphysema in hamsters was also prevented by TEI-5624 (1 mg/kg) administered intratracheally 7 hr after HSE administration (1 mg/kg). These results suggest that TEI-5624 and TEI-6344 may be useful therapeutic agents for the treatment of HLE-mediated diseases. PMID:8496803

Uejima, Y; Kokubo, M; Oshida, J; Kawabata, H; Kato, Y; Fujii, K

1993-05-01

323

Abstract. Objective and Design: Post transfusion infectious complications associated with allogeneic blood components  

E-print Network

leu- kocyte depleted by prestorage filtration, and whole blood leukocyte depleted by bedside-dependent manner during storage for 35 days of non-filtered whole blood, plasma-reduced whole blood, and SAGM blood YKL-40 accumulation was not reduced by bedside leukocyte depletion. Conclusion: YKL-40 appears

Price, Paul A.

324

[Morphodynamic analysis of the role of leukocytes in the microcirculation disorder of the cerebral cortex].  

PubMed

The aim of this work was to establish the reasons for the disturbances of microcirculation (no reflow) during various kinds of oxygen starvation of the brain. Using contact optics and video-recording, direct observations were performed and leukocyte and erythrocyte displacement was monitored in the blood vessels of rat cerebral cortex in vivo in normoxia, during the development of various kinds of oxygen deficiency up to the terminal periods of life of an organism. In normoxia leukocytes were shown to decelerate erythrocyte movement in capillaries and to cause the irregularity of the capillary blood flow. Using the color videofilm recording it was shown that at deep stages of brain oxygen deficiency the disturbances of microcirculation up to its complete cessation were explained by the occlusion of venous microvessels as a result of massive adhesion of leukocytes to the inner surface of venules and the smallest veins. The co-adhesion of leukocytes was also found to take place. The latter leads to the formation of large leukocyte aggregates in venous vessels, results in "no reflow" effect. Leukocyte aggregates in veins block the venous outflow from the brain and may become the immediate cause of organism death as a result of disruption of circulation and oxygen deficiency. PMID:14994593

Ivanov, K P; Mel'nikova, N N

2003-01-01

325

Metabolism of leukotriene B4 by activated human polymorphonuclear granulocytes.  

PubMed Central

Human polymorphonuclear granulocytes (PMNs) synthesize leukotriene B4 (LTB4) as a response of cell activation. Inactivation of the potent inflammatory mediator proceeds via omega-oxidation, resulting in the formation of 20-hydroxy- and 20-carboxy-LTB4. The main metabolite after stimulation with the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (FMLP) is 20-carboxy-LTB4, and after stimulation with the calcium ionophore A23187 is 20-hydroxy-LTB4. Differences in the LTB4 inactivation pathway were also observed when the catabolism of exogenously added LTB4 was analysed. In contrast to resting cells or cells preactivated with FMLP, prestimulation with the ionophore or with phorbol esters resulted in the inhibition of 20-carboxy-LTB4-generation. This decrease correlated with the reduction in specific [3H] LTB4-receptor expression. Studies with the non-penetrating diazonium salt of sulphanilic acid, which is known to interact with ectoenzymes, revealed that LTB4 is metabolized via receptor-mediated uptake. Our data suggest that the reduction in the amount of LTB4-receptor sites inhibits the conversion of 20-OH-LTB4 into 20-COOH-LTB4. PMID:2842254

Brom, J; Schonfeld, W; Konig, W

1988-01-01

326

Yersinia pseudotuberculosis Efficiently Escapes Polymorphonuclear Neutrophils during Early Infection  

PubMed Central

The human-pathogenic species of the Gram-negative genus Yersinia preferentially target and inactivate cells of the innate immune defense, suggesting that this is a critical step by which these bacteria avoid elimination and cause disease. In this study, bacterial interactions with dendritic cells, macrophages, and polymorphonuclear neutrophils (PMNs) in intestinal lymphoid tissues during early Yersinia pseudotuberculosis infection were analyzed. Wild-type bacteria were shown to interact mainly with dendritic cells, but not with PMNs, on day 1 postinfection, while avirulent yopH and yopE mutants interacted with PMNs as well as with dendritic cells. To unravel the role of PMNs during the early phase of infection, we depleted mice of PMNs by using an anti-Ly6G antibody, after which we could see more-efficient initial colonization by the wild-type strain as well as by yopH, yopE, and yopK mutants on day 1 postinfection. Dissemination of yopH, yopE, and yopK mutants from the intestinal compartments to mesenteric lymph nodes was faster in PMN-depleted mice than in undepleted mice, emphasizing the importance of effective targeting of PMNs by these Yersinia outer proteins (Yops). In conclusion, escape from interaction with PMNs due to the action of YopH, YopE, and YopK is a key feature of pathogenic Yersinia species that allows colonization and effective dissemination. PMID:24379291

Westermark, Linda; Fahlgren, Anna

2014-01-01

327

Ethanol enhances leukocyte-endothelial cell interactions in mesenteric venules.  

PubMed

In vivo studies have implicated neutrophils in the gastric mucosal injury produced by intraluminal administration of ethanol. However, in vitro studies indicate that ethanol inhibits various neutrophil functions such as adherence, chemotaxis, and degranulation. The aim of the present study was to assess whether ethanol, at clinically relevant concentrations, is proinflammatory in vivo. Ethanol (0.2, 1.0, 2.0, and 4.0%) was applied to the surface of the cat mesentery, and neutrophil adherence to venules (30 microns diam) and extravasation into the interstitium were quantitated using intravital microscopy. Hemodynamic parameters were also measured (venular diameter, red blood cell velocity, and leukocyte rolling velocity) or calculated (venular blood flow and wall shear stress). In this model ethanol produced a dose-dependent increase in neutrophil adherence and extravasation. The increase in leukocyte-endothelial cell interactions could not be attributed to alterations in hemodynamic factors. Pretreatment of animals with a monoclonal antibody (MoAb IB4) directed to the neutrophil CD11/CD18 adherence complex completely prevented the ethanol-induced neutrophil adherence and extravasation. Pretreatment with a leukotriene B4 (LTB4)-receptor antagonist (SC 41930) or a platelet-activating factor (PAF)-receptor antagonist (WEB 2170) did not alter the ethanol-induced neutrophil-endothelial interactions. We conclude that ethanol is proinflammatory at concentrations which may be achieved in the mucosal interstitium during acute alcohol intoxication. The ethanol-induced leukocyte adherence and extravasation is dependent on the expression of adhesive glycoproteins. The inflammatory mediators, PAF and LTB4, do not appear to play an important role in the leukocyte-endothelial cell interactions initiated by ethanol. PMID:2221068

Kvietys, P R; Perry, M A; Gaginella, T S; Granger, D N

1990-10-01

328

A mechanism for the antiinflammatory effects of corticosteroids: the glucocorticoid receptor regulates leukocyte adhesion to endothelial cells and expression of endothelial-leukocyte adhesion molecule 1 and intercellular adhesion molecule 1.  

PubMed Central

Corticosteroids are the preeminent antiinflammatory agents although the molecular mechanisms that impart their efficacy have not been defined. The endothelium plays a critical role in inflammation by directing circulating leukocytes into extravascular tissues by expressing adhesive molecules for leukocytes [e.g., endothelial-leukocyte adhesion molecule 1 (ELAM-1) and intercellular adhesion molecule 1 (ICAM-1)]. We therefore determined whether corticosteroids suppress inflammation by inhibiting endothelial expression of adhesion molecules for neutrophils (polymorphonuclear leukocytes). Preincubation of endothelial cells with endotoxin [lipopolysaccharide (LPS), 1 microgram/ml] led to a 4-fold increase in subsequent adherence of polymorphonuclear leukocytes (P < 0.0001, n = 10) to endothelial cells, an increase that was markedly attenuated when endothelial cells were treated with dexamethasone (IC50 < 1 nM, P < 0.0001, n = 6 or 7) during preincubation with LPS. Moreover, the steroid receptor agonist cortisol (10 microM), but not its inactive metabolite tetrahydrocortisol (10 microM), diminished LPS-induced endothelial cell adhesiveness. Further evidence that the action of dexamethasone was mediated through ligation of corticosteroid receptors [human glucocorticoid receptors (hGRs)] was provided by experiments utilizing the steroid antagonist RU-486. RU-486 (10 microM), which prevents translocation of ligated hGR to the nucleus by inhibiting dissociation of hGR from heat shock protein 90, completely aborted the effect of dexamethasone on adhesiveness of endothelial cells (P < 0.0005, n = 3). Treatment of endothelial cells with LPS (1 microgram/ml) stimulated transcription of ELAM-1, as shown by Northern blot analysis, and expression of membrane-associated ELAM-1 and ICAM-1, as shown by quantitative immunofluorescence (both P < 0.001, n = 9). Dexamethasone markedly inhibited LPS-stimulated accumulation of mRNA for ELAM-1 and expression of ELAM-1 and ICAM-1 (IC50 < 10 nM, both P < 0.001, n = 4-9); inhibition of expression by dexamethasone was reversed by RU-486 (both P < 0.005, n = 4-6). As in the adhesion studies, cortisol but not tetrahydrocortisol inhibited expression of ELAM-1 and ICAM-1 (both P < 0.005, n = 3 or 4). In contrast, sodium salicylate (1 mM) inhibited neither adhesion nor expression of these adhesion molecules. These studies suggest that antagonism by dexamethasone of endotoxin-induced inflammation is a specific instance of the general biological principle that the glucocorticoid receptor is a hormone-dependent regulator of transcription. Images PMID:1279685

Cronstein, B N; Kimmel, S C; Levin, R I; Martiniuk, F; Weissmann, G

1992-01-01

329

Dual Role of the Leukocyte Integrin ?M?2 in Angiogenesis.  

PubMed

Polymorphonuclear neutrophils (PMNs) and macrophages are crucial contributors to neovascularization, serving as a source of chemokines, growth factors, and proteases. ?M?2(CD11b/CD18) and ?L?2(CD11a/CD18) are expressed prominently and have been implicated in various responses of these cell types. Thus, we investigated the role of these ?2 integrins in angiogenesis. Angiogenesis was analyzed in wild-type (WT), ?M-knockout (?M (-/-)), and ?L-deficient (?L (-/-)) mice using B16F10 melanoma, RM1 prostate cancer, and Matrigel implants. In all models, vascular area was decreased by 50-70% in ?M (-/-) mice, resulting in stunted tumor growth as compared with WT mice. In contrast, ?L deficiency did not impair angiogenesis and tumor growth. The neovessels in ?M (-/-) mice were leaky and immature because they lacked smooth muscle cell and pericytes. Defective angiogenesis in the ?M (-/-) mice was associated with attenuated PMN and macrophage recruitment into tumors. In contrast to WT or the ?L (-/-) leukocytes, the ?M (-/-) myeloid cells showed impaired plasmin (Plm)-dependent extracellular matrix invasion, resulting from 50-75% decrease in plasminogen (Plg) binding and pericellular Plm activity. Surface plasmon resonance verified direct interaction of the ?MI-domain, the major ligand binding site in the ?2 integrins, with Plg. However, the ?LI-domain failed to bind Plg. In addition, endothelial cells failed to form tubes in the presence of conditioned medium collected from TNF-?-stimulated PMNs derived from the ?M (-/-) mice because of severely impaired degranulation and secretion of VEGF. Thus, ?M?2 plays a dual role in angiogenesis, supporting not only Plm-dependent recruitment of myeloid cells to angiogenic niches, but also secretion of VEGF by these cells. PMID:25261488

Soloviev, Dmitry A; Hazen, Stanley L; Szpak, Dorota; Bledzka, Kamila M; Ballantyne, Christie M; Plow, Edward F; Pluskota, Elzbieta

2014-11-01

330

Photochemically immobilized polymer coatings: effects on protein adsorption, cell adhesion, and leukocyte activation.  

PubMed

Amphiphilic chains of 4-benzoylbenzoic acid moieties and polymer were photochemically immobilized onto silicone rubber to ask whether the covalently coupled polymers would passivate the silicone rubber by inhibiting protein adsorption and subsequent cell adhesion and activation. Three groups of polymers were utilized: the hydrophilic synthetic polymers of polyacrylamide, polyethylene glycol, and polyvinylpyrrolidone; the glycosaminoglycan, hyaluronic acid; and poly(glycine-valine-glycine-valine-proline), a polypeptide derived from the sequence of elastin. Each coating variant decreased the adsorption of fibrinogen and immunoglobulin G compared to uncoated silicone rubber. All except the methoxy-polyethylene glycol coating nearly abolished fibroblast growth, but none of the coating variants inhibited monocyte or polymorphonuclear leukocyte adhesion. Interleukin-1beta, interleukin-1 receptor antagonist, and tumor necrosis factor-alpha secretion by leukocytes were not statistically different between any of the coating variants and uncoated silicone rubber. However, the methoxy-polyethylene glycol and elastin-based polypeptide coatings, which supported the highest numbers of adherent monocytes, also elicited the lowest levels of proinflammatory cytokine secretion. When these in vitro data were collectively evaluated, the coating that most effectively passivated silicone rubber was the polypeptide derived from elastin. PMID:10591132

Defife, K M; Hagen, K M; Clapper, D L; Anderson, J M

1999-01-01

331

FURTHER OBSERVATIONS ON THE BEHAVIOR OF STAPHYLOCOCCI WITHIN HUMAN LEUKOCYTES  

PubMed Central

A specific serum factor was required for rapid phagocytosis of pathogenic staphylococci by human polymorphonuclear leukocytes when the ingestion process was studied in siliconed glass systems and the concentrations of staphylococci were maintained at low levels. In contrast to certain other microbes, the resistance to phagocytosis which characterized pathogenic staphylococci was relative, and phagocytosis was readily accomplished when large populations of staphylococci were present in the test system. A factor promoting phagocytosis was present in eight of eight normal adult sera. In contrast, the sera of twenty-eight of thirty normal rabbits did not promote phagocytosis. Serum obtained from 2 rabbits maintained in the rabbit colony for several months acquired the ability to opsonize pathogenic staphylococci. The phagocytosis-promoting factor was almost completely removed by prior absorption of test sera with the homologous strain. The factor was incompletely removed by absorption with heterologous strains of pathogenic staphylococci and was not significantly reduced by absorption with coagulase-negative staphylococci or unrelated microorganisms. Present evidence suggests that the factor promoting phagocytosis is a thermostable opsonin. While the activity of heated serum could not be restored by the addition of small amounts of fresh serum or complement, the addition of large amounts of complement partially restored opsonic activity. Incubation of staphylococci in fresh serum prior to heat inactivation did not reduce subsequent phagocytosis, further suggesting the heat stability of the phagocytosis-promoting factor. Preliminary studies correlating the presence of antistaphylococcal hemagglutinins and phagocytosis-promoting factor in certain sera suggest that the two factors were not necessarily related. The phagocytosis of staphylococci in fresh human serum was inhibited by the addition of fresh or inactivated rabbit serum. Further studies on the nature of such inhibition are in progress. Once ingestion was accomplished, coagulase-positive staphylococci consistently survived in significant numbers within the cytoplasm of human granulocytes. Coagulase-negative staphylococci appeared to be destroyed within the leukocyte and could not be recultured from the cytoplasm following 3 to 4 hours of intracellular residence. PMID:13742745

Rogers, David E.; Melly, Marian Ann

1960-01-01

332

Preparation and evaluation of a /sup 99m/Tc-SnF2 colloid kit for leukocyte labeling  

SciTech Connect

Stannous fluoride colloid (SFC) kits for instant radiolabeling with 99mTc were prepared and evaluated for suitability as a leukocyte radiolabeling agent. Technetium-99m labeling for kits stored at -15/degree/C for up to 3 mo was greater than 95% as determined by instant thin layer chromatography while colloid particles of 1-3 microns were measured by electron microscope for these preparations. Canine leukocyte preparations labeled with (/sup 99m/Tc)SFC and characterized by triple density gradients of metrizamide in plasma demonstrated an 83% leukocyte association. Analysis of labeled cell preparation for up to 3 hr demonstrated label stability. Labeled leukocytes, when readministered in normal dogs, demonstrated bi-exponential blood clearance with uptake and subsequent clearance from lung. There was increasing uptake of labeled leukocytes by the liver until steady state was achieved. Furthermore, when whole blood samples were analyzed by the triple density gradient method, an increasing monocyte-to-granulocyte ratio was observed to occur with time. By 3 hr 95% of the whole blood activity was associated with the leukocyte fraction. Dogs in which a 24-hr sterile abscess was created demonstrated elevated blood-pool activity as compared to control with localization of the labeled cells at inflammatory sites within 3 hr following cell readministration.

Hirsch, J.I.; Tatum, J.L.; Fratkin, M.J.; Apostolides, D.L.; Quint, R.I.

1989-07-01

333

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 ...

334

[Effect of proteflazid on TLRs expression by mononuclear leukocytes of peripheral blood and epithelial cells of mucous membranes and skin in patients with herpes-associated erythema multiforme and erythema annulare centrifugum].  

PubMed

The article reports survey data on 23 patients with erythemas, including 19 patients with herpes-associated erythema multiforme (HAEM) and 4 patients with Darier's erythema annulare centrifugum (DEAC). Patients in the initial state (baseline) and after two weeks of therapy with proteflazid were characterized by measuring the levels of Toll-like receptor (TLR) expression in peripheral blood mononuclear cells (PBMC) and in epithelial cells of the throat and the skin. The TLR expression in PBMC and skin was assessed by flow cytometry with monoclonal antibodies (ICA) (Caltag Laboratories, USA; Hycult Biotech, Netherlands) against relevant antigens. In addition, patients were also characterized by the content of subpopulations of lymphocytes expressing surface markers CD3, CD4, CD8, CD16, CD21, CD23, CD72, CD25, and HLA-DR in the peripheral blood, which was measured by flow cytometry. The therapy with proteflazid in patients with both HAEM and DEAC led to normalization of the level of both T-cell and B-cell immunity, which was manifested by an increase in the total number of lymphocytes, CD3+, CD4+, CD21+, and CD72+. Measurements of the dynamics of TLR expression in the course of immunotherapy showed an increase in the number of TLR 2, 3, 4, 7, 8, and 9 in PBMC (which was especially pronounced for TLR2) and in epithelium of the pharyngeal mucosa and skin (increased expression of TLR3, 7, and 9). PMID:24800523

Sorokina, E V; Akhmatova, N K; Skhodova, S A

2014-01-01

335

Serine protease autotransporters from Shigella flexneri and pathogenic Escherichia coli target a broad range of leukocyte glycoproteins.  

PubMed

The serine protease autotransporters of Enterobacteriaceae (SPATEs) are secreted by pathogenic Gram-negative bacteria through the autotransporter pathway. We previously classified SPATE proteins into two classes: cytotoxic (class 1) and noncytotoxic (class 2). Here, we show that Pic, a class 2 SPATE protein produced by Shigella flexneri 2a, uropathogenic and enteroaggregative Escherichia coli strains, targets a broad range of human leukocyte adhesion proteins. Substrate specificity was restricted to glycoproteins rich in O-linked glycans, including CD43, CD44, CD45, CD93, CD162 (PSGL-1; P-selectin glycoprotein ligand 1), and the surface-attached chemokine fractalkine, all implicated in leukocyte trafficking, migration, and inflammation. N-terminal sequencing of proteolytic products revealed Pic (protease involved in colonization) cleavage sites to occur before Thr or Ser residues. The purified carbohydrate sLewis-X implied in inflammation and malignancy inhibited cleavage of PSGL-1 by Pic. Exposure of human leukocytes to purified Pic resulted in polymorphonuclear cell activation, but impaired chemotaxis and transmigration; Pic-treated T cells underwent programmed cell death. We also show that the Pic-related protease Tsh/Hbp, implicated in extraintestinal infections, exhibited a spectrum of substrates similar to those cleaved by Pic. In the guinea pig keratoconjunctivitis model, a Shigella pic mutant induced greater inflammation than its parent strain. We suggest that the class-2 SPATEs represent unique immune-modulating bacterial virulence factors. PMID:21768350

Ruiz-Perez, Fernando; Wahid, Rezwanul; Faherty, Christina S; Kolappaswamy, Krishnan; Rodriguez, Liliana; Santiago, Araceli; Murphy, Ebony; Cross, Alan; Sztein, Marcelo B; Nataro, James P

2011-08-01

336

Serine protease autotransporters from Shigella flexneri and pathogenic Escherichia coli target a broad range of leukocyte glycoproteins  

PubMed Central

The serine protease autotransporters of Enterobacteriaceae (SPATEs) are secreted by pathogenic Gram-negative bacteria through the autotransporter pathway. We previously classified SPATE proteins into two classes: cytotoxic (class 1) and noncytotoxic (class 2). Here, we show that Pic, a class 2 SPATE protein produced by Shigella flexneri 2a, uropathogenic and enteroaggregative Escherichia coli strains, targets a broad range of human leukocyte adhesion proteins. Substrate specificity was restricted to glycoproteins rich in O-linked glycans, including CD43, CD44, CD45, CD93, CD162 (PSGL-1; P-selectin glycoprotein ligand 1), and the surface-attached chemokine fractalkine, all implicated in leukocyte trafficking, migration, and inflammation. N-terminal sequencing of proteolytic products revealed Pic (protease involved in colonization) cleavage sites to occur before Thr or Ser residues. The purified carbohydrate sLewis-X implied in inflammation and malignancy inhibited cleavage of PSGL-1 by Pic. Exposure of human leukocytes to purified Pic resulted in polymorphonuclear cell activation, but impaired chemotaxis and transmigration; Pic-treated T cells underwent programmed cell death. We also show that the Pic-related protease Tsh/Hbp, implicated in extraintestinal infections, exhibited a spectrum of substrates similar to those cleaved by Pic. In the guinea pig keratoconjunctivitis model, a Shigella pic mutant induced greater inflammation than its parent strain. We suggest that the class-2 SPATEs represent unique immune-modulating bacterial virulence factors. PMID:21768350

Ruiz-Perez, Fernando; Wahid, Rezwanul; Faherty, Christina S.; Kolappaswamy, Krishnan; Rodriguez, Liliana; Santiago, Araceli; Murphy, Ebony; Cross, Alan; Sztein, Marcelo B.; Nataro, James P.

2011-01-01

337

Intracytoplasmic Ultrastructures in Peripheral Blood Cells in Lupus Erythematosus.  

National Technical Information Service (NTIS)

Buffy coats of peripheral blood of different manifestations of lupus erythematosus were studied by electron microscopy for the existence of viruslike intracytoplasmic tubular structures. They could always be demonstrated in the blood leukocytes of systemi...

G. Metz, J. Metz

1974-01-01

338

Measurement of residual leukocytes in platelet concentrate units.  

PubMed

Leukocyte contamination of various blood products is implicated in febrile transfusion reactions, in alloimmunization phenomena and immunosuppression, and in the transmission of viral infections. The biggest difficulty in interpreting published reports lies in the methods used to quantify residual leukocytes. As automated cell counters are not precise and sensitive enough to detect low cell counts, we compared two techniques: i) the classical method using the Nageotte haemocytometer; and ii) fluorescent staining with propidium iodide (PI). We performed these two techniques to evaluate the residual white cell count in 41 platelet units collected by cytapheresis. The difference in results obtained with the two techniques was borderline significant (P = 0.05). To evaluate the accuracy of both techniques, we added a known number of lymphocytes to platelet concentrates that had been filtered three times. There was a good correlation between the values obtained using the two techniques (r > 0.99). However, though the difference between the known value and PI results was not significant, the difference for Nageotte cell counter results was significant (P < 0.0001). From a practical point of view, the PI fluorescent technique is simple, rapid, and easy to carry out. Our results demonstrate the reliability of the PI technique for the evaluation of residual leukocytes. PMID:8166394

Fizet, D; Bouzgarrou, R; Piquet, Y; Cristol, G; Vezon, G

1993-01-01

339

Non-specific cytotoxic activity of teleost leukocytes.  

PubMed

The existence of lymphoid cells with "natural" killer activity in mammals and birds has been known for some time. Several previous reports have demonstrated similar activity in carp and catfish kidney leukocytes. However, the activity previously reported was directed towards established mammalian cell lines. In this report we confirm the existence of spontaneous killer activity in other species of teleosts, including salmonids. This spontaneous cytotoxic activity is directed towards several established teleost, as well as mammalian, cell lines. Cytotoxic activity appears to be optimal at 20 degrees C in an 8 hour 51Cr release assay. The RTG-2, AS, GS and EPC cell lines of teleost origin are susceptible to lysis by kidney, spleen, and blood leukocytes of Salmo salar, Salmo gairdneri, and Notemigonus crysoleucas. Furthermore, the susceptibility of the RTG-2 and AS teleost cell lines to killing by kidney leukocytes of both S. salar and S. gairdneri was significantly enhanced by preinfection of the target cells with infectious pancreatic necrosis virus. PMID:3996708

Moody, C E; Serreze, D V; Reno, P W

1985-01-01

340

Glycobiology of leukocyte trafficking in inflammation.  

PubMed

To fulfill their potential, leukocytes must be able to exit the vasculature and reach the site of inflammation within the tissue. This process of leukocyte extravasation is a tightly regulated sequence of events that is governed by a host of cell adhesion molecules, cytokines, chemokines and lipid mediators. Of major importance to this process and the function of many of the proteins and lipids involved is the posttranslational modification of these moieties by glycosylation. The glycosylation process is coordinated by multiple enzymes that add and remove saccharides to/from glycan structures on proteins and lipids, resulting in a unique molecular signature that affords specificity to the molecules involved in leukocyte recruitment. This review will discuss how glycosylation impacts the function of these key molecules involved in the recruitment of leukocytes during inflammation and the function of specific lectins (carbohydrate-binding proteins) that have a role in leukocyte trafficking. PMID:25258391

Wright, Rachael D; Cooper, Dianne

2014-12-01

341

Effects of fluvastatin on leukocyte-endothelial cell adhesion in hypercholesterolemic rats.  

PubMed

The overall objective of this study was to determine whether peroral treatment with the 3-hydroxy-3-methyl-glutaryl coenzyme A (HMG-CoA) reductase inhibitor fluvastatin influences the leukocyte-endothelial cell adhesion (LECA) observed in postcapillary venules of hypercholesterolemic rats. Rats were fed either normal chow or a chow supplemented with 1% cholesterol for 10 days. Leukocyte adherence and extravasation, leukocyte rolling velocity, red blood cell velocity, and vessel diameter were monitored in mesenteric venules superfused with either 100 nmol/L platelet-activating factor (PAF) or 20 nmol/L leukotriene B4 (LTB4). Hypercholesterolemic rats exhibited an exaggerated LECA response compared with their normocholesterolemic counterparts. In hypercholesterolemic rats, treatment with fluvastatin significantly attenuated the leukocyte-adherence responses to PAF and LTB4 as well as the leukocyte emigration response to LTB4. Fluvastatin treatment also inhibited the PAF- and LTB4-induced reductions in leukocyte rolling velocity. These findings indicate that fluvastatin blunts the inflammatory responses elicited in post-capillary venules by lipid mediators. PMID:9301630

Kimura, M; Kurose, I; Russell, J; Granger, D N

1997-08-01

342

Telomere shortening in leukocyte subpopulations in depression  

PubMed Central

Background Telomere shortening is a normal age-related process. However, premature shortening of telomeres in leukocytes – as has been reported in depression – may increase the risk for age-related diseases. While previous studies investigated telomere length in peripheral blood mononuclear cells (PBMCs) as a whole, this study investigated specific changes in the clonal composition of white blood cells of the adaptive immune system (CD4+ helper and CD8+ cytotoxic T lymphocytes, and CD20+ B lymphocytes). Methods Forty-four females with a history of unipolar depression were investigated and compared to fifty age-matched female controls. Telomere lengths were compared between three groups: 1) individuals with a history of depression but currently no clinically relevant depressive symptoms, 2) individuals with a history of depression with relevant symptoms of depression, and 3) healthy age-matched controls. Telomere length was assessed using quantitative fluorescence in situ hybridization (qFISH). Results Both groups with a history of unipolar depression (with and without current depressive symptoms) showed significantly shorter telomeres in all three lymphocyte subpopulations. The effect was stronger in CD8+ and CD20+ cells than in CD4+ cells. Individuals with a history of depression and with (without) current symptoms exhibited a CD8+ telomere length shortening corresponding to an age differential of 27.9 (25.3) years. Conclusions A history of depression is associated with shortened telomeres in the main effector populations of the adaptive immune system. Shorter telomeres seem to persist in individuals with lifetime depression independently of the severity of depressive symptoms. CD8+ cytotoxic T cells and CD20+ B cells seem to be particularly affected in depression. The total number of depressive episodes did not influence telomere length in the investigated adaptive immune cell populations. PMID:24996455

2014-01-01

343

Nanowire array chips for molecular typing of rare trafficking leukocytes with application to neurodegenerative pathology  

NASA Astrophysics Data System (ADS)

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 the BBB and infiltrate into the CNS. The 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 the CSF may represent important sources to investigate immune-to-brain interactions or diagnose and monitor neurodegenerative diseases. Due to the paucity of trafficking leucocytes in the 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 phenotypes. A 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 for potentially diagnosing and monitoring the pathogenesis of neurodegenerative diseases by allowing an effective hematological analysis of the CSF from patients.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 the BBB and infiltrate into the CNS. The 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 the CSF may represent important sources to investigate immune-to-brain interactions or diagnose and monitor neurodegenerative diseases. Due to the paucity of trafficking leucocytes in the 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 phenotypes. A 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 for potentially diagnosing and monitoring the pathogenesis of neurodegenerative diseases by allowing an effective hematological analysis of the CSF from patients. Electro

Kwak, Minsuk; Kim, Dong-Joo; Lee, Mi-Ri; Wu, Yu; Han, Lin; Lee, Sang-Kwon; Fan, Rong

2014-05-01

344

Usefulness of indium-111-oxine-labeled leukocyte scintigraphy in diagnosis of inflammation associated with chronic aortic dissection  

Microsoft Academic Search

Background: Patients with chronic aortic dissection require monitoring for indications of disease progression. In present study, inflammation\\u000a adjacent to associated aortic wall was evaluated by indium-111-oxine-labeled leukocyte scintigraphy, scince inflammation of\\u000a the blood vessel wall often associates with progression of chronic aortic dissection.Methods and Results: Fifteen patients with aortic dissection underwent indium-111-oxine-labeled leukocyte scintigraphy. Seven showed positive\\u000a images at sites

Keiko Takahashi; Mitsumasa Ohyanagi; Hitoshi Naruse; Miho Masai; Tadaaki Iwasaki; Minoru Fukuchi; Takashi Miyamoto

2001-01-01

345

A Genome-Wide Gene Expression Signature of Environmental Geography in Leukocytes of Moroccan Amazighs  

Microsoft Academic Search

The different environments that humans experience are likely to impact physiology and disease susceptibility. In order to estimate the magnitude of the impact of environment on transcript abundance, we examined gene expression in peripheral blood leukocyte samples from 46 desert nomadic, mountain agrarian and coastal urban Moroccan Amazigh individuals. Despite great expression heterogeneity in humans, as much as one third

Youssef Idaghdour; John D. Storey; Sami J. Jadallah; Greg Gibson

2008-01-01

346

TRAIL-coated leukocytes that kill cancer cells in the circulation  

E-print Network

to neutralize circulating tumor cells that enter blood with the potential to form new metastases. drug delivery tumor can intravasate into the peripheral circulation as circulating tumor cells (CTCs) (2, 3TRAIL-coated leukocytes that kill cancer cells in the circulation Michael J. Mitchell, Elizabeth

Schaffer, Chris B.

347

Microfluidics for in vitro biomimetic shear stress-dependent leukocyte adhesion assays.  

PubMed

Recruitment of leukocytes from blood to tissues is a multi-step process playing a major role in the activation of inflammatory responses. Tethering and rolling of leukocytes along the vessel wall, followed by arrest and transmigration through the endothelium result from chemoattractant-dependent signals, inducing adhesive and migratory events. Shear forces exerted by the blood flow on leukocytes induce rolling via selectin-mediated interactions with endothelial cells and increase the probability of leukocytes to engage their chemokine receptors, facilitating integrin activation and consequent arrest. Flow-derived shear forces generate mechanical stimuli concurring with biochemical signals in the modulation of leukocyte-endothelial cell interactions. In the last few years, a host of in vitro studies have clarified the biochemical adhesion cascade and the role of shear stress in leukocyte extravasation. The limitation of the static environment in Boyden devices has been overcome both by the use of parallel-plate flow chambers and by custom models mimicking the in vivo conditions, along with widespread microfluidic approaches to in vitro modeling. These devices create an in vitro biomimetic environment where the multi-step transmigration process can be imaged and quantified under mechanical and biochemical controlled conditions, including fluid dynamic settings, channel design, materials and surface coatings. This paper reviews the technological solutions recently proposed to model, observe and quantify leukocyte adhesion behavior under shear flow, with a final survey of high-throughput solutions featuring multiple parallel assays as well as thorough and time-saving statistical interpretation of the experimental results. PMID:23200903

Bianchi, Elena; Molteni, Raffaella; Pardi, Ruggero; Dubini, Gabriele

2013-01-18

348

The Role of Platelets in Leukocyte Recruitment in Chronic Contact Hypersensitivity Induced by Repeated Elicitation  

PubMed Central

Platelets have been shown to be important in inflammation, but their role in chronic allergic dermatitis remains unclear. To investigate the role of platelets in a mouse model of chronic contact hypersensitivity induced by repeated elicitation, mice were sensitized and repeatedly elicited in ears with hapten, with or without platelet depletion, by administering antiplatelet antibody or busulfan. Ear thickness, leukocyte infiltration, serum IgE, and scratching behavior significantly decreased in thrombocytopenic mice. cDNA microarray of ear tissue showed reduced gene expression associated with Th2 lymphocytes. Flow cytometry showed increased P-selectin expression on platelets and an increased number of platelet-leukocyte aggregates in blood of repeatedly elicited mice, compared with sham-sensitized mice. In thrombocytopenic mice, inflammation was restored by platelet infusion, which was blocked by platelets from P-selectin-deficient mice or by pretreating platelets with anti-P-selectin antibody. Moreover, injection of activated platelet supernatant into ears led to increased leukocyte infiltration, which was blocked by pretreating platelets with antiplatelet compounds or neutralizing several chemokines in the platelet supernatant. These results suggest that platelets induce leukocyte recruitment into skin by forming platelet-leukocyte aggregates via P-selectin in blood and secreting chemokines at inflamed sites. Therefore, controlling platelet activity may be useful for treatment of chronic allergic dermatitis. PMID:17525269

Tamagawa-Mineoka, Risa; Katoh, Norito; Ueda, Eiichiro; Takenaka, Hideya; Kita, Masakazu; Kishimoto, Saburo

2007-01-01

349

Role of p38 MAPK in LPS induced pro-inflammatory cytokine and chemokine gene expression in equine leukocytes  

Microsoft Academic Search

Endotoxemia occurs when bacterial lipopolysaccharide (LPS) in the blood induces a dysregulated inflammatory response, resulting in circulatory shock and multi-organ failure. Laminitis is a common complication in endotoxemic horses and is frequently the reason for humane euthanasia of these cases. Blood leukocytes are a principal target of LPS in endotoxemia leading to activation of multiple signal transduction pathways involved in

Laura E. Neuder; Jamie M. Keener; Rachael E. Eckert; Jennifer C. Trujillo; Samuel L. Jones

2009-01-01

350

TRAIL-coated leukocytes that kill cancer cells in the circulation.  

PubMed

Metastasis through the bloodstream contributes to poor prognosis in many types of cancer. Mounting evidence implicates selectin-based adhesive interactions between cancer cells and the blood vessel wall as facilitating this process, in a manner similar to leukocyte trafficking during inflammation. Here, we describe a unique approach to target and kill colon and prostate cancer cells in the blood that causes circulating leukocytes to present the cancer-specific TNF-related apoptosis inducing ligand (TRAIL) on their surface along with E-selectin adhesion receptor. This approach, demonstrated in vitro with human blood and also in mice, mimics the cytotoxic activity of natural killer cells and increases the surface area available for delivery of the receptor-mediated signal. The resulting "unnatural killer cells" hold promise as an effective means to neutralize circulating tumor cells that enter blood with the potential to form new metastases. PMID:24395803

Mitchell, Michael J; Wayne, Elizabeth; Rana, Kuldeepsinh; Schaffer, Chris B; King, Michael R

2014-01-21

351

Sour cherry seed kernel extract increases heme oxygenase-1 expression and decreases representation of CD3+ TNF-?+ and CD3+IL-8+ subpopulations in peripheral blood leukocyte cultures from type 2 diabetes patients.  

PubMed

The present study evaluates a hypothesis that sour cherry (Prunus cerasus) seed extracts (SCE) modulate CD3+ T lymphocyte activity in ways predictive of potential for uses of SCE in management of inflammatory diseases. Peripheral blood mononuclear cells (PBMC) from 12 type 2 diabetes (T2DM) patients and eight healthy control subjects were cultured 24 h with 100 ng/ml lipopolysaccharide (LPS) to increase inflammatory signaling and co-incubated with 0.5-100 µg/ml SCE. Cultures were evaluated by two-color flow cytometry for percent representation of CD3+ IL8+ and CD3+TNF-? cells which express interleukin-8 (IL-8), and tumor necrosis factor-?, (TNF-?+) respectively, and by enzyme-linked immunoassay for lymphocyte-associated heme oxygenase-1 (HO-1, known to be induced by SCE). SCE dosage ranges of 0.5-100 µg/ml in cell cultures significantly suppressed LPS-increased CD3+TNF-?+ and CD3+IL8+ representation from all participants (p?

Mahmoud, Fadia F; Al-Awadhi, Rana; Haines, David D; Dashti, Ali; Dashti, Hussain; Al-Ozairi, Ebaa; Bak, Istvan; Tosaki, Arpad

2013-05-01

352

Optimizing Viable Leukocyte Sampling from the Female Genital Tract for Clinical Trials: An International Multi-Site Study  

PubMed Central

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

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

353

Cardiopulmonary Bypass during Cardiac Surgery Modulates Systemic Inflammation by Affecting Different Steps of the Leukocyte Recruitment Cascade  

PubMed Central

Background It is known that the use of a cardiopulmonary bypass (CPB) during cardiac surgery leads to leukocyte activation and may, among other causes, induce organ dysfunction due to increased leukocyte recruitment into different organs. Leukocyte extravasation occurs in a cascade-like fashion, including capturing, rolling, adhesion, and transmigration. However, the molecular mechanisms of increased leukocyte recruitment caused by CPB are not known. This clinical study was undertaken in order to investigate which steps of the leukocyte recruitment cascade are affected by the systemic inflammation during CPB. Methods We investigated the effects of CPB on the different steps of the leukocyte recruitment cascade in whole blood from healthy volunteers (n?=?9) and patients undergoing cardiac surgery with the use of cardiopulmonary bypass (n?=?7) or in off-pump coronary artery bypass-technique (OPCAB, n?=?9) by using flow chamber experiments, transmigration assays, and biochemical analysis. Results CPB abrogated selectin-induced slow leukocyte rolling on E-selectin/ICAM-1 and P-selectin/ICAM-1. In contrast, chemokine-induced arrest and transmigration was significantly increased by CPB. Mechanistically, the abolishment of slow leukocyte rolling was due to disturbances in intracellular signaling with reduced phosphorylation of phospholipase C (PLC) ?2, Akt, and p38 MAP kinase. Furthermore, CPB induced an elevated transmigration which was caused by upregulation of Mac-1 on neutrophils. Conclusion These data suggest that CPB abrogates selectin-mediated slow leukocyte rolling by disturbing intracellular signaling, but that the clinically observed increased leukocyte recruitment caused by CPB is due to increased chemokine-induced arrest and transmigration. A better understanding of the underlying molecular mechanisms causing systemic inflammation after CPB may aid in the development of new therapeutic approaches. PMID:23029213

Rossaint, Jan; Berger, Christian; Van Aken, Hugo; Scheld, Hans H.; Zahn, Peter K.; Rukosujew, Andreas; Zarbock, Alexander

2012-01-01

354

Response of laying hens to feeding low-protein amino acid-supplemented diets under high ambient temperature: performance, egg quality, leukocyte profile, blood lipids, and excreta pH  

NASA Astrophysics Data System (ADS)

An experiment was conducted to determine whether, by using a low-protein amino acid-supplemented diet, the health status, stress response, and excreta quality could be improved without affecting the productive performance of heat-stressed laying hens. The requirements for egg production, egg mass, and feed conversion ratio were also estimated using second-order equations and broken-line regression. A total of 150 Lohmann Selected Leghorn (LSL-Lite) hens were divided randomly into five groups of 30 with five replicates of six hens. The hens were raised for an 8-week period (52 to 60 weeks) in wire cages situated in high ambient temperature in an open-sided housing system. The five experimental diets (ME; 2,720 kcal/kg) varied according to five crude protein (CP) levels: normal-CP diet (control, 16.5 % CP) and low-CP diets containing 15.0, 13.5, 12.0, or 10.5 % CP. All experimental diets were supplemented with crystalline amino acids at the levels sufficient to meet their requirements. The results showed that under high temperature conditions, all productive performance and egg quality parameters in the birds fed with 15.0, 13.5, and 12.0 % CP diets were similar to those of birds fed with control diet (16.5 % CP), whereas feeding 10.5 % CP diet significantly decreased egg production and egg mass. Estimations of requirements were of 13.93 and 12.77 % CP for egg production, 14.62 and 13.22 % CP for egg mass, and 12.93 and 12.26 % CP for feed conversion ratio using quadratic and broken-line models, respectively. Egg yolk color index, blood triglyceride level, and excreta acidity were also significantly higher in birds fed with 12.0 and 10.5 % CP diets compared with those of control birds. The heterophil to lymphocyte ratio, as a stress indicator, was significantly decreased by 15.0, 13.5, and 12 % CP diets. On the basis of our findings, reducing dietary CP from 16.5 to 12.0 % and supplementing the diets with the essential amino acids showed merit for improving the stress response and excreta quality while maintaining acceptable production performance from laying hens under high ambient temperature conditions.

Torki, Mehran; Mohebbifar, Ahmad; Ghasemi, Hossein Ali; Zardast, Afshin

2014-07-01

355

[The noninvasive technique of analysis of aggregation activity of thrombocytes, leukocytes and erythrocytes].  

PubMed

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

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

356

Evaluation of polymorphonuclear cell and monocyte functions in Leishmania infantum-infected dogs  

Microsoft Academic Search

Leishmania infantum is the causative agent of canine leishmaniasis in the Mediterranean area. The aspects of the canine immune response which may explain the chronic severe disease in these animals have not been well investigated. Therefore, in this work we have evaluated the parasite killing ability by circulating polymorphonuclear (PMN) cells and monocytes in dogs with spontaneous leishmaniasis and in

O. Brandonisio; M. Panunzio; S. M. Faliero; L. Ceci; A. Fasanella; V. Puccini

1996-01-01

357

Killing of Escherichia coli by human polymorphonuclear leucocytes in the presence of Bacteroides fragilis  

Microsoft Academic Search

The inhibitory effect of Bacteroides fragilis on the in vitro killing of Escherichia coli by polymorphonuclear leucocytes was studied with two pairs of E coli and B fragilis isolated from human wound infections. Both B fragilis strains behaved similarly: they inhibited the killing of one E coli strain, while the killing of the other E coli strain was not affected.

W A Vel; F Namavar; A M Verweij-van Vught; A N Pubben; D M MacLaren

1985-01-01

358

Generation of slow-reacting substance (leukotrienes) by endotoxin and lipid A from human polymorphonuclear granulocytes.  

PubMed Central

Leukotrienes were released from human polymorphonuclear granulocytes on incubation with endotoxins and lipid A. The analysis was performed by their smooth muscle contracting properties, reversed phase high-pressure liquid chromatography and radioimmunoassay for leukotrienes C4 and D4. The active component of the lipopolysaccharides seems to be the lipid A portion. PMID:6490085

Bremm, K D; Konig, W; Spur, B; Crea, A; Galanos, C

1984-01-01

359

Generation of slow-reacting substance (leukotrienes) by endotoxin and lipid A from human polymorphonuclear granulocytes.  

PubMed

Leukotrienes were released from human polymorphonuclear granulocytes on incubation with endotoxins and lipid A. The analysis was performed by their smooth muscle contracting properties, reversed phase high-pressure liquid chromatography and radioimmunoassay for leukotrienes C4 and D4. The active component of the lipopolysaccharides seems to be the lipid A portion. PMID:6490085

Bremm, K D; König, W; Spur, B; Crea, A; Galanos, C

1984-10-01

360

Uptake, Transport, and Delivery of Antimicrobial Agents by Human Polymorphonuclear Neutrophils  

Microsoft Academic Search

Polymorphonuclear neutrophils (PMN) are attracted to sites of infection. They have the potential to deliver antimicrobial agents to these sites if the agents enter the cells and do not alter migration. Penicillin G did not enter cells and was not transported by PMN. We found that azithromycin, ciprofloxacin, levofloxacin, moxi- floxacin, and telithromycin were concentrated in PMN and transported toward

Gerald L. Mandell; Elizabeth Coleman

2001-01-01

361

Comparison of venous and capillary differential leukocyte counts using a standard hematology analyzer and a novel microfluidic impedance cytometer.  

PubMed

Capillary blood sampling has been identified as a potentially suitable technique for use in diagnostic testing of the full blood count (FBC) at the point-of-care (POC), for which a recent need has been highlighted. In this study we assess the accuracy of capillary blood counts and evaluate the potential of a miniaturized cytometer developed for POC testing. Differential leukocyte counts in the normal clinical range from fingerprick (capillary) and venous blood samples were measured and compared using a standard hematology analyzer. The accuracy of our novel microfluidic impedance cytometer (MIC) was then tested by comparing same-site measurements to those obtained with the standard analyzer. The concordance between measurements of fingerprick and venous blood samples using the standard hematology analyzer was high, with no clinically relevant differences observed between the mean differential leukocyte counts. Concordance data between the MIC and the standard analyzer on same-site measurements presented significantly lower leukocyte counts determined by the MIC. This systematic undercount was consistent across the measured (normal) concentration range, suggesting that an internal correction factor could be applied. Differential leukocyte counts obtained from fingerprick samples accurately reflect those from venous blood, which confirms the potential of capillary blood sampling for POC testing of the FBC. Furthermore, the MIC device demonstrated here presents a realistic technology for the future development of FBC and related tests for use at the site of patient care. PMID:23028467

Hollis, Veronica S; Holloway, Judith A; Harris, Scott; Spencer, Daniel; van Berkel, Cees; Morgan, Hywel

2012-01-01

362

Basic Fibroblast Growth Factor (bFGF, FGF-2) Potentiates Leukocyte Recruitment to Inflammation by Enhancing Endothelial Adhesion Molecule Expression  

PubMed Central

Basic fibroblast growth factor (bFGF, FGF-2) is a potent angiogenic factor and endothelial cell mitogen. Although bFGF levels are increased in chronically inflamed tissue, its role in inflammation is unclear. We investigated the effect of bFGF on acute dermal inflammation and the recruitment of monocytes, T cells, and neutrophils. Leukocyte recruitment to inflamed sites was quantified with radiolabeled leukocytes. Intradermal injection of bFGF in rats did not induce leukocyte recruitment or inflammation. However, the recruitment of leukocytes to inflammation induced by tumor necrosis factor-?, interferon-?, C5a, or a delayed hypersensitivity reaction was enhanced by bFGF by 55 to 132% (P < 0.05). Either acute or prolonged bFGF treatment of dermal sites had this effect. The potentiating effect of bFGF on leukocyte recruitment was also seen in joints. There was no associated modulation of vascular permeability, blood flow, or angiogenesis in the sites by bFGF. However, the expression of the endothelial cell adhesion molecules (CAMs) for leukocytes, P-selectin, E-selectin, and ICAM-1, was significantly up-regulated in the inflamed tissue by bFGF, as quantified by radiolabeled anti-CAM antibody binding in vivo. Thus, although not directly proinflammatory, bFGF synergistically potentiates inflammatory mediator-induced leukocyte recruitment, at least in part, by enhancing CAM up-regulation on endothelium. PMID:16507899

Zittermann, Sandra I.; Issekutz, Andrew C.

2006-01-01

363

A Novel Mutation in Leukocyte Adhesion Deficiency Type II/CDGIIc.  

PubMed

Leukocyte adhesion deficiencies (LAD) are autosomal recessive immunodeficiency syndromes characterized by severe and recurrent bacterial infections, impaired wound healing and leukocytosis. Block in different steps in the leukocyte adhesion cascade causes different types of leukocyte adhesion deficiencies, LAD type I, II and III. In LAD type II, the rolling phase of the leukocyte adhesion cascade is affected due to mutations in the specific fucose transporter GFTP (GDP fucose transporter), causing defect in the biosynthesis of selectin ligands on leukocytes. Thus this syndrome is also called congenital disorder of glycosylation IIc (CGDIIc). LAD II/CGDIIc is very rare and has been diagnosed in nine children to date. Fever, leukocytosis, typical dysmorphic features, growth, psychomotor retardation and the Bombay blood group, are characteristic findings in patients. Here, we describe two Turkish siblings with a novel mutation in GFTP. They both have the characteristic features of the syndrome. The older sibling died of severe bacterial pneumonia at the age of 3 years. The younger sibling, diagnosed at the age of 3 months, responded to high dose oral fucose supplementation. Secundum atrial septal defect which was not described in previously reported patients, but present in both of our patients, may primarily related to the defect in fucosylation. PMID:25239688

Cagdas, Deniz; Y?lmaz, Mustafa; Kandemir, Nurgün; Tezcan, Ilhan; Etzioni, Amos; Sanal, Ozden

2014-11-01

364

A prospective study of leukocyte telomere length and risk of renal cell carcinoma  

PubMed Central

Background It has been hypothesized that genomic instability related to telomere dysfunction may contribute to carcinogenesis. There is some evidence from case-control studies suggesting that short leukocyte telomere length (TL) may be associated with an increased risk of renal cell carcinoma (RCC); however, this association has not been investigated prospectively. Methods We conducted a nested case-control study (209 cases, 410 controls) of RCC risk in relation to pre-diagnostic leukocyte TL in the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial. Odds ratios (OR) and 95% confidence intervals (CI) were estimated using conditional logistic regression. Results Leukocyte TL was not significantly associated with future risk of RCC (highest quartile vs. lowest: OR=0.8, 95% CI=0.5–1.5; Ptrend=0.6). Analyses stratified by sex, age, and time from blood collection to RCC diagnosis were similarly null. Conclusions The results of this study, to our knowledge the first prospective investigation of its kind, do not support an association between pre-diagnostic leukocyte TL and risk of RCC. Impact In contrast to some earlier reports, our findings add to the evidence that leukocyte TL is not a biomarker of risk related to the etiology of RCC. PMID:23513041

Hofmann, Jonathan N.; Lan, Qing; Cawthon, Richard; Hosgood III, H. Dean; Shuch, Brian; Moore, Lee E.; Rothman, Nathaniel; Chow, Wong-Ho; Purdue, Mark P.

2013-01-01

365

Exposure to sodium fluoride produces signs of apoptosis in rat leukocytes.  

PubMed

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

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

366

Three-dimensional modulation of cortical plasticity during pseudopodial protrusion of mouse leukocytes.  

PubMed

Leukocytes can rapidly migrate virtually within any substrate found in the body at speeds up to 100 times faster than mesenchymal cells that remain firmly attached to a substrate even when migrating. To understand the flexible migration strategy utilized by leukocytes, we experimentally investigated the three-dimensional modulation of cortical plasticity during the formation of pseudopodial protrusions by mouse leukocytes isolated from blood. The surfaces of viable leukocytes were discretely labeled with fluorescent beads that were covalently conjugated with concanavalin A receptors. The movements of these fluorescent beads were different at the rear, central, and front surfaces. The beads initially present on the rear and central dorsal surfaces of the cell body flowed linearly toward the rear peripheral surface concomitant with a significant collapse of the cell body in the dorsal-ventral direction. In contrast, those beads initially on the front surface moved into a newly formed pseudopodium and exhibited rapid, random movements within this pseudopodium. Bead movements at the front surface were hypothesized to have resulted from rupture of the actin cytoskeleton and detachment of the plasma membrane from the actin cytoskeletal cortex, which allowed leukocytes to migrate while being minimally constrained by a substrate. PMID:23939049

Miyoshi, Hiromi; Tsubota, Ken-ichi; Hoyano, Takamasa; Adachi, Taiji; Liu, Hao

2013-09-01

367

Soluble CD40 ligand accumulates in stored blood components, primes neutrophils through CD40, and is a potential cofactor in the development of transfusion-related acute lung injury  

PubMed Central

Transfusion-related acute lung injury (TRALI) is a form of posttransfusion acute pulmonary insufficiency that has been linked to the infusion of biologic response modifiers (BRMs), including antileukocyte antibodies and lipids. Soluble CD40 ligand (sCD40L) is a platelet-derived proinflammatory mediator that accumulates during platelet storage. We hypothesized that human polymorpho-nuclear leukocytes (PMNs) express CD40, CD40 ligation rapidly primes PMNs, and sCD40L induces PMN-mediated cytotoxicity of human pulmonary microvascular endothelial cells (HMVECs). Levels of sCD40L were measured in blood components and in platelet concentrates (PCs) implicated in TRALI or control PCs that did not elicit a transfusion reaction. All blood components contained higher levels of sCD40L than fresh plasma, with apheresis PCs evidencing the highest concentration of sCD40L followed by PCs from whole blood, whole blood, and packed red blood cells (PRBCs). PCs implicated in TRALI reactions contained significantly higher sCD40L levels than control PCs. PMNs express functional CD40 on the plasma membrane, and recombinant sCD40L (10 ng/mL-1 ?g/mL) rapidly (5 minutes) primed the PMN oxidase. Soluble CD40L promoted PMN-mediated cytotoxicity of HMVECs as the second event in a 2-event in vitro model of TRALI. We concluded that sCD40L, which accumulates during blood component storage, has the capacity to activate adherent PMNs, causing endothelial damage and possibly TRALI in predisposed patients. PMID:16772606

Khan, Samina Yasmin; Kelher, Marguerite R.; Heal, Joanna M.; Blumberg, Neil; Boshkov, Lynn K.; Phipps, Richard; Gettings, Kelly F.; McLaughlin, Nathan J.; Silliman, Christopher C.

2006-01-01

368

In vivo temporal and spatial profile of leukocyte adhesion and migration after experimental traumatic brain injury in mice  

PubMed Central

Background Leukocytes are believed to be involved in delayed cell death following traumatic brain injury (TBI). However, data demonstrating that blood-borne inflammatory cells are present in the injured brain prior to the onset of secondary brain damage have been inconclusive. We therefore investigated both the interaction between leukocytes and the cerebrovascular endothelium using in vivo imaging and the accumulation of leukocytes in the penumbra following experimentally induced TBI. Methods Experimental TBI was induced in C57/Bl6 mice (n = 42) using the controlled cortical impact (CCI) injury model, and leukocyte-endothelium interactions (LEI) were quantified using both intravital fluorescence microscopy (IVM) of superficial vessels and 2-photon microscopy of cortical vessels for up to 14 h post-CCI. In a separate experimental group, leukocyte accumulation and secondary lesion expansion were analyzed in mice that were sacrificed 15 min, 2, 6, 12, 24, or 48 h after CCI (n = 48). Finally, leukocyte adhesion was blocked with anti-CD18 antibodies, and the effects on LEI and secondary lesion expansion were determined 16 (n = 12) and 24 h (n = 21), respectively, following TBI. Results One hour after TBI leukocytes and leukocyte-platelet aggregates started to roll on the endothelium of pial venules, whereas no significant LEI were observed in pial arterioles or in sham-operated mice. With a delay of >4 h, leukocytes and aggregates did also firmly adhere to the venular endothelium. In deep cortical vessels (250 ?m) LEIs were much less pronounced. Transmigration of leukocytes into the brain parenchyma only became significant after the tissue became necrotic. Treatment with anti-CD18 antibodies reduced adhesion by 65%; however, this treatment had no effect on secondary lesion expansion. Conclusions LEI occurred primarily in pial venules, whereas little or no LEI occurred in arterioles or deep cortical vessels. Inhibiting LEI did not affect secondary lesion expansion. Importantly, the majority of migrating leukocytes entered the injured brain parenchyma only after the tissue became necrotic. Our results therefore suggest that neither intravascular leukocyte adhesion nor the migration of leukocytes into cerebral tissue play a significant role in the development of secondary lesion expansion following TBI. PMID:23448240

2013-01-01

369

Small amounts of venous gas embolism cause delayed impairment of endothelial function and increase polymorphonuclear neutrophil infiltration.  

PubMed

Gas bubbles from decompression and gas embolization lead to endothelial dysfunction and mechanical injury in the pig, rabbit and lamb. In the study presented here, 0.01 ml air/min/kg was infused through a catheter into the jugular vein in 12 rabbits for 60 min. The endothelial response was measured using tension measurements in the blood vessel wall, and morphological changes where quantified using light microscopy and image processing. Percent lung water content was calculated and used to estimate the severity of pulmonary oedema. The infusion led to a significant decrease in the acetylcholine-mediated endothelial-dependent vasodilatation in the pulmonary artery 6 h after the infusion (6-h group, n = 6). A decrease in substance-P-mediated endothelial-dependent vasodilatation was also detected. No changes where seen in a group of rabbits examined 1 h after infusion (l-h group, n=6). The impaired endothelial-dependent vasodilatation caused by the bubbles is probably biochemical in origin, since no visible changes were seen in the endothelial layer. A significant increase in polymorphonuclear neutrophils was observed in the 6-h group compared to the l-h group. This study demonstrates that small numbers of bubbles, corresponding to "silent bubbles", lead to an impairment of the endothelial-dependent vasoactive response. PMID:11990728

Nossum, Vibeke; Hjelde, Astrid; Brubakk, Alf O

2002-01-01

370

Next-generation qPCR for the high-throughput measurement of gene expression in multiple leukocyte subsets.  

PubMed

Clinical studies of gene expression are increasingly using the whole blood, peripheral blood mononuclear cells, and leukocyte subsets involved in the innate and adaptive immune responses. However, the small amount of RNA available in the clinical setting is a limitation for commonly used methods such as quantitative polymerase chain reactions (qPCR) and microarrays. Our aim was to design 96 gene assays to simultaneously measure gene expression in the whole blood and seven leukocyte subsets using a new-generation qPCR method--high-throughput nanofluidic reverse transcription qPCR (HT RT-qPCR). The leukocyte subset purity was 94% to 98% for seven subsets and was less for the ?? T-cell receptor subset (80%). The HT RT-qPCR replicate sample measurements were highly reproducible (r = 0.997, p < 2.2 × 10(-16)), and the ??Ct values from HT RT-qPCR correlated significantly with those from qPCR. The control genes were differentially expressed across the eight leukocyte subsets in the control subjects (p = 1.3 × 10(-5), analysis of variance). Two analytical methods, absolute and relative, gave concordant results and were significantly correlated (p = 1.9 × 10(-9)). HT RT-qPCR permits the rapid, reproducible, and quantitative measurement of multiple transcripts using minimal sample amounts. The protocol described yielded leukocyte subsets of high purity and identified two analytic methods for use. PMID:23690294

Adamski, Mateusz G; Li, Yan; Wagner, Erin; Yu, Hua; Seales-Bailey, Chloe; Soper, Steven A; Murphy, Michael; Baird, Alison E

2013-10-01

371

Interaction of Aspergillus fumigatus Spores with Human Leukocytes and Serum  

PubMed Central

Serum was necessary for optimal phagocytosis of Aspergillus fumigatus spores by human leukocytes, and its opsonic capacity was greatly diminished by heat inactivation (56 C, 30 min). A germination assay, described in this report, was developed to study the fate of phagocytized spores. After incubation for 3 hr with normal leukocytes and serum, spores ingested by peripheral blood neutrophils and monocytes remained viable. Since we had previously found that myeloperoxidase (MPO), a lysosomal enzyme of human neutrophils and monocytes, exerted fungicidal activity against Candida albicans when combined with H2O2 and chloride or iodide, the effects of these substances on A. fumigatus spores were examined. Spore viability was not impaired by MPO alone, H2O2 alone, or KI alone, but high concentrations of KI and H2O2 in combination caused marked inhibition of subsequent germination. MPO imparted fungicidal activity to concentrations of KI and H2O2 that lacked any effect in its absence. NaCl, in combination with MPO and H2O2, was far less effective than the iodide salt against A. fumigatus. The relative ineffectiveness of chloride in this system could underly the apparent inability of human neutrophils to kill ingested A. fumigatus spores, despite their competence to kill C. albicans. Images PMID:16557740

Lehrer, Robert I.; Jan, Ronald G.

1970-01-01

372

Reperfusion injury after 24 hours cold storage of donor hearts. Possible role of leukocyte activation.  

PubMed

The role of neutrophils on the development of reperfusion injury was evaluated in canine hearts. Reperfusion was performed with a closed reperfusion system. 18 dog hearts were arrested by a single dose of National Institute of Health cardioplegia and stored cold (0.5 degrees C) for 24 hours. Thereafter they were randomly attached to one of the following groups. Group I: reperfusion with whole blood (n = 6), group II: reperfusion with low-leukocyte blood (n = 6), group III: reperfusion with low-leukocyte blood plus phenyl-isopropyl adenosine (PIA) (n = 6). Hearts in group I showed an increase of high energy phosphates (HEP) within the first 30 minutes, and a significant decrease during the subsequent 30 minutes. Concomitantly the initially soft hearts turned into stone hearts. In group II and III a significant increase of HEP was seen during the entire reperfusion. After 60 minutes of reperfusion myocardial ATP levels in group III were significantly higher than in group I. All hearts in group III were soft after reperfusion. It is concluded that reperfusion injury appears to be dependent upon the presence and activation of neutrophils, which can be partially prevented by reperfusion with low-leukocyte blood and more so by the simultaneous administration of PIA. PMID:2349552

Möllhoff, T; Sukehiro, S; Van Aken, H; Flameng, W

1990-04-01

373

Cuprophan reuse and intradialytic changes of lung diffusion capacity and blood gasses  

Microsoft Academic Search

Cuprophan reuse and intradialytic changes of lung diffusion capacity and arterial blood gasses. The changes in arterial blood gas, pulmonary function tests, leukocyte counts and complement activition were evaluated during first use and subsequent reuse of cuprophan dialyzers. The dialysate buffer was bicarbonate. Reuse of cuprophan dialyzers significantly attenuated the fall in leukocyte counts and the rise in C3a des

Raymond C Vanholder; Romain A Pauwels; Johan F Vandenbogaerde; Herman H Lamont; Marcel E Van Der Straeten; Severin M Ringoir

1987-01-01

374

CD66 carcinoembryonic antigens mediate interactions between Opa-expressing Neisseria gonorrhoeae and human polymorphonuclear phagocytes  

Microsoft Academic Search

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

Scott D. Gray-Owen; Christoph Dehio; Anja Haude; Fritz Grunert; Thomas F. Meyer

1997-01-01

375

agr-Dependent Interactions of Staphylococcus aureus USA300 with Human Polymorphonuclear Neutrophils  

Microsoft Academic Search

The emergence of serious infections due to community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has fueled interest in the contributions of specific staphylococcal virulence factors to clinical disease. To assess the contributions of agr-dependent factors to the fate of organisms in polymorphonuclear neutrophils (PMN), we examined the consequences for organism and host cells of feeding PMN with wild-type CA-MRSA (LAC) or CA-MRSA

Yun Yun Pang; Jamie Schwartz; Matthew Thoendel; Laynez W. Ackermann; Alexander R. Horswill; William M. Nauseef

2010-01-01

376

Heterotropic modulation of selectin affinity by allosteric antibodies affects leukocyte rolling  

PubMed Central

Selectins are a family of adhesion receptors designed for efficient leukocyte tethering to the endothelium under shear. As a key property to resist premature bond disruption, selectin adhesiveness is enhanced by tensile forces that promote the conversion of a bent into an extended conformation of the N-terminal lectin and EGF-like domains. Conformation-specific antibodies have been invaluable in deciphering the activation mechanism of integrins, but similar reagents are not available for selectins. Here we show that the anti-human L-selectin monoclonal antibodies DREG-55 and LAM1.5 but not DREG-56, -200 or LAM1.1 heterotropically modulate adhesion presumably by stabilizing the extended receptor conformation. Force-free affinity assays, flow chamber and microkinetic studies reveal a ligand-specific modulation of L-selectin affinity by DREG-55 mAb, resulting in a dramatic decrease of rolling velocity under flow. Furthermore, secondary tethering of polymorphonuclear cells was blocked by DREG-200 but significantly boosted by DREG-55 mAb. The results emphasize the need for a new classification for selectin antibodies and introduce the new concept of heterotropic modulation of receptor function. PMID:24431230

Riese, Sebastian B; Kuehne, Christian; Tedder, Thomas F; Hallmann, Rupert; Hohenester, Erhard; Buscher, Konrad

2013-01-01

377

Modeling leukocyte-leukocyte non-contact interactions in a lymph node.  

PubMed

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

Gritti, Nicola; Caccia, Michele; Sironi, Laura; Collini, Maddalena; D'Alfonso, Laura; Granucci, Francesca; Zanoni, Ivan; Chirico, Giuseppe

2013-01-01

378

Halo sign on indium-111 leukocyte scan in gangrenous cholecystitis  

SciTech Connect

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.

Bauman, J.M.; Boykin, M.; Hartshorne, M.F.; Cawthon, M.A.; Landry, A.J.

1986-02-01

379

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

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