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Lymphocyte-monocyte defect associated with anergy and recurrent infections  

PubMed Central

Mechanisms of cellular immunity were studied in an anergic female with recurrent bacterial infections. Lymphocytes transformed and produced cytotoxin in the presence of phytohaemagglutinin, but elaborated only migration inhibitory factor (MIF) in response to specific antigen. Monocytes isolated from the patient's peripheral blood were unresponsive to autologous MIF and to homologous MIF from two of three healthy adults. Humoral immunity, complement and neutrophil bactericidal activity were intact. The lymphocyte-monocyte defects found in this patient suggest that they were responsible for the anergy and predisposition to bacterial infections.

Louie, J. S.; Goldberg, L. S.



HIV-1 Infection and First Line ART Induced Differential Responses in Mitochondria from Blood Lymphocytes and Monocytes: The ANRS EP45 "Aging" Study  

PubMed Central

Background The ANRS EP45 “Aging” study investigates the cellular mechanisms involved in the accelerated aging of HIV-1 infected and treated patients. The data reported focus on mitochondria, organelles known to be involved in cell senescence. Methods 49 HIV-1 infected patients untreated with antiretroviral therapy, together with 49 seronegative age- and sex-matched control subjects and 81 HIV-1 infected and treated patients, were recruited by 3 AIDS centres (Marseille, Montpellier, Nice; France;, NCT01038999). In more than 88% of treated patients, the viral load was <40 copies/ml and the CD4+ cell count was >500/mm3. ROS (reactive oxygen species) production and ??m (inner membrane potential) were measured by flow cytometry in blood lymphocytes and monocytes (functional parameters). Three mitochondrial network quantitative morphological parameters were computed using confocal microscopy and image analysis. Three PBMC mitochondrial proteins (porin and subunits 2 and 4 of cytochrome C oxidase encoded by mtDNA or nuclear DNA, respectively) were analysed by western blotting. Results Quantitative changes in PBMC mitochondrial proteins were not induced by either HIV-1 infection or ART. Discriminant analysis integrating functional (ROS production and ??m) or morphological (network volume density, fragmentation and branching) parameters revealed HIV-1 infection and ART differential effects according to cell type. First line ART tended to rescue lymphocyte mitochondrial parameters altered by viral infection, but induced slight changes in monocytes. No statistical difference was found between the effects of three ART regimens on mitochondrial parameters. Correlations between functional parameters and viral load confirmed the damaging effects of HIV-1 in lymphocyte mitochondria. Conclusions In patients considered to be clinically stable, mitochondria exhibited functional and morphological modifications in PBMCs resulting from either direct or indirect effects of HIV-1 infection (lymphocytes), or from first line ART (monocytes). Together with other tissue impairments, these changes may contribute to global aging. Trial Registration NCT01038999 NCT01038999

Perrin, Sophie; Cremer, Jonathan; Roll, Patrice; Faucher, Olivia; Menard, Amelie; Reynes, Jacques; Dellamonica, Pierre; Naqvi, Alissa; Micallef, Joelle; Jouve, Elisabeth; Tamalet, Catherine; Solas, Caroline; Pissier, Christel; Arnoux, Isabelle; Nicolino-Brunet, Corine; Espinosa, Leon; Levy, Nicolas; Kaspi, Elise; Robaglia-Schlupp, Andree; Poizot-Martin, Isabelle; Cau, Pierre



IFN-?-activated lymphocytes boost nitric oxide production in grass carp monocytes/macrophages.  


It is well known that IFN-? is a prime activator of nitric oxide (NO) production by monocytes/macrophages in mammals and fish. In parallel, whether IFN-?-activated lymphocytes are associated with NO production remains unclear. In this study, grass carp monocytes/macrophages and lymphocytes from head kidney were isolated and effects of recombinant grass carp IFN-? (rgcIFN-?) on NO releases by these two cell populations were determined. Results showed that rgcIFN-? time- and dose-dependently increased NO production by monocytes/macrophages but not lymphocytes, which are consistent with the findings in mammals. Interestingly, rgcIFN-? displayed a greater stimulation on NO production in the co-cultures of monocytes/macrophages and lymphocytes when compared with that in the culture of monocytes/macrophages alone. Furthermore, the media harvested from rgcIFN-?-treated lymphocytes were effective in boosting NO release in monocytes/macrophages. These data suggest that secretions from rgcIFN-?-treated lymphocytes may be involved in the NO release by monocytes/macrophages. To address this hypothesis, effect of rgcIFN-? on the gene expression of inflammatory cytokines in grass carp lymphocytes was examined, showing that it consistently stimulated the mRNA expression of grass carp TNF-? and IL-1? but not IFN-?. Furthermore, treatment of rgcIFN-? combined with recombinant grass carp IL-1? (rgcIL-1?) induced a NO production by monocytes/macrophages, which was significantly higher than those induced by either cytokine alone. It provides the evidence that the cytokines secreted by the activated lymphocytes may facilitate the NO production by monocytes/macrophages. Taken together, our findings point out a new mechanism for the involvement of IFN-?-activated lymphocytes in the NO production by monocytes/macrophages in fish. This knowledge not only strengthens the role of IFN-? in immune system but also provides the evidence for the existence of a close relationship between lymphocytes and monocytes/macrophages in fish. PMID:24056277

Yang, Kun; Zhang, Shengnan; Chen, Danyan; Zhang, Anying; Wang, Xinyan; Zhou, Hong



Lymphocyte supernatant-induced human monocyte tumoricidal activity: dependence on the presence of gamma-interferon.  


Recently, gamma-interferon (IFN-gamma) has been shown to be have the capacity to activate macrophages in several murine and human systems. The studies reported here were undertaken to determine the identity of the lymphokine responsible for activation of human monocytes to a tumoricidal state. Macrophage-activating factor (MAF) activity was assessed using a 24-h 51Cr release assay with human monocytes as effector cells and K-562 targets. Stimulated lymphocyte supernatants were produced by stimulation of peripheral blood mononuclear cells with concanavalin A in serum-free media. Interferon was detected in an antiviral assay. Four lines of evidence lead to the conclusion that MAF and IFN-gamma are identical in this system: (a) fractionation of stimulated lymphocyte supernatants by adsorption chromatography, followed by anion or cation exchange chromatography (Mono-S, Mono-Q columns), resulted in nearly identical elution profiles of MAF and IFN activities. All of the individual fractions containing MAF activity were found to contain IFN in amounts corresponding to MAF activity. (b) Monoclonal antibody specific for IFN-gamma neutralized the ability of stimulated lymphocyte supernatants to induce human monocyte tumoricidal activity. This antibody also neutralized the MAF activity of purified IFN-gamma but not alpha-interferon. (c) The biological MAF activity of activated lymphocyte supernatants and IFN-gamma were similar. Dilution versus MAF activity for IFN-gamma and stimulated lymphocyte supernatants exhibited identical slopes. Lymphocyte supernatants and IFN-gamma demonstrated similar MAF activity on three effector cells: monocytes, in vitro-differentiated macrophages, and dexamethasone-differentiated macrophages. (d) Analysis of supernatants produced by five antigen-stimulated human T-cell clones demonstrated coordinate production of MAF and IFN. These results provide compelling evidence for support of the concept that IFN-gamma is the major human lymphokine capable of inducing monocyte-macrophage tumoricidal activity. PMID:3921233

Sadlik, J R; Hoyer, M; Leyko, M A; Horvat, R; Parmely, M; Whitacre, C; Zwilling, B; Rinehart, J J



Differential down-regulation of CD95 or CD95L in chronically HIV-infected cells of monocytic or lymphocytic origin: cellular studies and molecular analysis by quantitative competitive RT-PCR  

Microsoft Academic Search

We analysed the expression of CD95\\/CD95L in two widely used models for studying the cellular effects of chronic infection with human immunodeficiency virus type 1 (HIV-1), i.e. ACH-2 cells, derived from the lymphocytic cell line A301, and U1, derived from monocytic U937 cells. A301 and ACH-2 mounted the same amount of plasma membrane CD95, while U1 had a consistent decrease

Marcello Pinti; Jessica Pedrazzi; Francesca Benatti; Veronica Sorrentino; Cira Nuzzo; Valeria Cavazzuti; Priscilla Biswas; Daniela Nicoleta Petrusca; Cristina Mussini; Bruno De Rienzo; Andrea Cossarizza



Immunomodulatory effects of cardiotrophin-1 on in vitro cytokine production of monocytes & CD4+ T-lymphocytes  

PubMed Central

Background & objectives: In congestive heart failure (CHF), increased concentrations of several cytokines including cardiotrophin-1 (CT-1) and immunactivation are found. This study was performed to evaluate whether CT-1 can induce in vitro cytokines in monocytes and CD4+ T-lymphocytes of healthy volunteers. Methods: The study was performed in vitro to see whether CT-1 can modulate monocyte or CD4+ T-lymphocyte interleukin (IL)-1?, -2, -4, -5, -10, interferon ? (IFN?), and tumour necrosis factor ? (TNF?) expression by flow cytometry following stimulation with CT-1 alone or together with lipopolysaccharide (LPS) or phorbol myristate acetate (PMA)/ionomycine (iono). Results: CT-1 increased the number of TNF? and IL-1? positive monocytes. LPS induced IL-10, TNF?, and IL-1? in monocytes but only IL-2 in CD4+ T-lymphocytes, whereas PMA/iono induced all cytokines besides IL-5 in monocytes and IL-1? in CD4+ T-lymphocytes. In LPS activated monocytes, CT-1 induced a concentration-dependent reduction in the number of TNF? positive monocytes. After LPS activation, CT-1 decreased the number of CD4+ lymphocytes positive for IL-2, IL-4, and IL-5. In addition, following PMA/iono stimulation, CT-1 initiated a concentration-dependent decrease of CD4+ T-lymphocytes positive for TNF?, IL-4, IL-5, and IL-10. Interpretation & conclusions: The present data show that in vitro CT-1 can activate monocytes and modulate cytokine production of activated CD4+ T-lymphocytes. We speculate that CT-1 may at least be partly responsible for immunactivation in CHF.

Fritzenwanger, Michael; Jung, Christian; Franz, Marcus; Foerster, Martin; Figulla, Hans R.



Peripheral blood lymphocyte/monocyte ratio at diagnosis and survival in classical Hodgkin's lymphoma  

PubMed Central

Background Lymphopenia and tumor-associated macrophages are negative prognostic factors for survival in classical Hodgkin’s lymphoma. We, therefore, studied whether the peripheral blood absolute lymphocyte count/absolute monocyte count ratio at diagnosis affects survival in classical Hodgkin’s lymphoma. Design and Methods We studied 476 consecutive patients with classical Hodgkin’s lymphoma followed at the Mayo Clinic from 1974 to 2010. Receiver operating characteristic curves and area under the curve were used to determine cut-off values for the absolute lymphocyte count/absolute monocyte count ratio at diagnosis, while proportional hazards models were used to compare survival based on the absolute lymphocyte count/absolute monocyte count ratio at diagnosis. Results The median follow-up period was 5.6 years (range, 0.1–33.7 years). An absolute lymphocyte count/absolute monocyte count ratio at diagnosis of 1.1 or more was the best cut-off value for survival with an area under the curve of 0.91 (95% confidence interval, 0.86 to 0.96), a sensitivity of 90% (95% confidence interval, 85% to 96%) and specificity of 79% (95% confidence interval, 73% to 88%). Absolute lymphocyte count/absolute monocyte count ratio at diagnosis was an independent prognostic factor for overall survival (hazard ratio, 0.18; 95% confidence interval, 0.08 to 0.38, P<0.0001); lymphoma-specific survival (hazard ratio, 0.10; 95% confidence interval, 0.04 to 0.25, P<0.0001); progression-free survival (hazard ratio, 0.35; 95% confidence interval, 0.18 to 0.66, P<0.002) and time to progression (hazard ratio, 0.27; 95% confidence interval, 0.17 to 0.57, P<0.0006). Conclusions The ratio of absolute lymphocyte count/absolute monocyte count at diagnosis is an independent prognostic factor for survival and provides a single biomarker to predict clinical outcomes in patients with classical Hodgkin’s lymphoma.

Porrata, Luis F.; Ristow, Kay; Colgan, Joseph P.; Habermann, Thomas M.; Witzig, Thomas E.; Inwards, David J.; Ansell, Stephen M.; Micallef, Ivana N.; Johnston, Patrick B.; Nowakowski, Grzegorz S.; Thompson, Carrie; Markovic, Svetomir N.



Peripheral blood monocyte-to-lymphocyte ratio at study enrollment predicts efficacy of the RTS,S malaria vaccine: analysis of pooled phase II clinical trial data  

PubMed Central

Background RTS,S is the most advanced candidate malaria vaccine but it is only partially protective and the causes of inter-individual variation in efficacy are poorly understood. Here, we investigated whether peripheral blood monocyte-to-lymphocyte ratios (ML ratio), previously shown to correlate with clinical malaria risk, could account for differences in RTS,S efficacy among phase II trial participants in Africa. Methods Of 11 geographical sites where RTS,S has been evaluated, pre-vaccination ML ratios were only available for trial participants in Kilifi, Kenya (N?=?421) and Lambarene, Gabon (N?=?189). Using time to first clinical malaria episode as the primary endpoint we evaluated the effect of accounting for ML ratio on RTS,S vaccine efficacy against clinical malaria by Cox regression modeling. Results The unadjusted efficacy of RTS,S in this combined dataset was 47% (95% confidence interval (CI) 26% to 62%, P <0.001). However, RTS,S efficacy decreased with increasing ML ratio, ranging from 67% (95% CI 64% to 70%) at an ML ratio of 0.1 to 5% (95% CI -3% to 13%) at an ML ratio of 0.6. The statistical interaction between RTS,S vaccination and ML ratio was still evident after adjustment for covariates associated with clinical malaria risk in this dataset. Conclusion The results suggest that stratification of study participants by ML ratio, easily measured from full differential blood counts before vaccination, might help identify children who are highly protected and those that are refractory to protection with the RTS,S vaccine. Identifying causes of low vaccine efficacy among individuals with high ML ratio could inform strategies to improve overall RTS,S vaccine efficacy. Trial registration ClinicalTrials.Gov numbers NCT00380393 and NCT00436007



Granulocyte-monocyte-colony-stimulating factor augments the cytotoxic capacity of lymphocytes and monocytes in antibody-dependent cellular cytotoxicity  

Microsoft Academic Search

Human peripheral blood mononuclear cells (lymphocytes and monocytes) were preincubated for 0–24 h with human recombinant granulocyte-monocyte-colony-stimulating factor (GM-CSF) and used as effector cells in an 18 h antibody-dependent cellular cytotoxicity (ADCC) assay with SW948 (a human colorectal carcinoma cell line) as target cells and mAb 17-1A. A significant increase in the lytic capability was noted after 0.5–2 h of

G. Masucci; P. Wersäll; P. Ragnhammar; H. Mellstedt



Effects of increasing docosahexaenoic acid intake in human healthy volunteers on lymphocyte activation and monocyte apoptosis  

PubMed Central

Dietary intake of long-chain n-3 polyunsaturated fatty acids (n-3 PUFA) has been reported to decrease several markers of lymphocyte activation and modulate monocyte susceptibility to apoptosis. However most human studies examined the combined effect of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) using relatively high daily amounts of n-3 PUFA. The present study investigated the effects of increasing doses of DHA added to the regular diet of human healthy volunteers on lymphocyte response to tetradecanoylphorbol acetate (TPA) plus ionomycin activation, and on monocyte apoptosis induced by oxidized LDL (oxLDL). Eight subjects were supplemented with increasing daily doses of DHA (200, 400, 800 and 1600mg) in a triacylglycerol form containing DHA as the only PUFA, for two weeks each dose. DHA intake dose-dependently increased the proportion of DHA in mononuclear cell phospholipids, the augmentation being significant after 400mg DHA/day. The TPA plus ionomycin-stimulated IL-2 mRNA level started to increase after ingestion of 400mg DHA/day, with a maximum after 800mg intake, and was positively correlated (P<0.003) with DHA enrichment in cell phospholipids. The treatment of monocytes by oxLDL before DHA supplementation drastically reduced mitochondrial membrane potential as compared with native LDL treatment. OxLDL apoptotic effect was significantly attenuated after 400mg DHA/day and the protective effect was maintained throughout the experiment, although to a lesser extent at higher doses. The present results show that supplementation of the human diet with low DHA dosages improves lymphocyte activability. It also increases monocyte resistance to oxLDL-induced apoptosis, which may be beneficial in the prevention of atherosclerosis.

Mebarek, Saida; Ermak, Natalia; Benzaria, Amal; Vicca, Stephanie; Dubois, Madeleine; Nemoz, Georges; Laville, Martine; Lacour, Bernard; Vericel, Evelyne; Lagarde, Michel; Prigent, Annie-France



Differential down-regulation of CD95 or CD95L in chronically HIV-infected cells of monocytic or lymphocytic origin: cellular studies and molecular analysis by quantitative competitive RT-PCR.  


We analysed the expression of CD95/CD95L in two widely used models for studying the cellular effects of chronic infection with human immunodeficiency virus type 1 (HIV-1), i.e. ACH-2 cells, derived from the lymphocytic cell line A301, and U1, derived from monocytic U937 cells. A301 and ACH-2 mounted the same amount of plasma membrane CD95, while U1 had a consistent decrease in CD95 expression. Using different antibodies, we failed to detect the plasma membrane form of its ligand, CD95L, but we could see the intracellular presence of that molecule in A301 cells and, to a lesser extent, in ACH-2 cells, but not in U937 or U1 cells. To confirm the cytofluorimetric data and quantify the expression of CD95L at the RNA level, we developed a quantitative competitive RT-PCR assay. The HUT78 cell line had about 50,000 copies mRNA/1000 cells, three times more after induction with a phorbol ester and ionomycin. ACH-2 expressed about 400- (basal) or 10- (induced) fold less CD95L mRNA than the parental cell line A301; U937 and U1 were below the limit of detection. In cells of lymphoid origin (ACH-2) chronic HIV infection inhibits the expression of CD95L, the phenomenon occurring at the transcriptional level. In cells of monocytic origin (U1) the infection decreases the plasma membrane expression of CD95. This suggests that HIV could trigger different anti-apoptotic strategies which likely depend upon the cell line which is infected. In monocytic cells which act as a viral reservoir, the expression of the molecule whose binding triggers apoptosis decreases, while in lymphoid cells, capable of exerting cytotoxicity, the expression of a molecule which induces apoptosis is reduced. PMID:10481067

Pinti, M; Pedrazzi, J; Benatti, F; Sorrentino, V; Nuzzo, C; Cavazzuti, V; Biswas, P; Petrusca, D N; Mussini, C; De Rienzo, B; Cossarizza, A



T-lymphocyte induction of human monocyte angiotensin converting enzyme (ACE) is not dependent upon T-lymphocyte proliferation  

SciTech Connect

Human peripheral blood monocytes cultured in serum free media for seven days show a basal activity of the ectoenzyme ACE which is augmented 2-3 times by the presence of autologous peripheral blood T-lymphocytes. Since these two cell types are also involved in autologous mixed lymphocyte reaction if serum is present, the authors compared the ability of T-cells to stimulate ACE activity in the presence or absence of proliferation (measured by /sup 3/H-thymidine incorporation). By the seventh day, cultures with 5% AB/sup +/ serum showed significant increase in proliferation but no increase in ACE activity compared to the serum free cultures. Even higher proliferation rate achieved by co-culturing T-lymphocytes with allogeneic monocytes did not increase ACE production; on the contrary, ACE activity remained at the basal level. Monocyte-T-cell co-cultures stimulated with increasing concentrations of ConA or PHA showed dose dependent increases in proliferation but parallel decreases in ACE activity. Addition of soluble antigen (Candida albicans) also enhanced proliferation but not ACE synthesis. They conclude that T-lymphocyte induction of monocyte ACE is a result of cooperation between autologous cells which is not dependent upon T-cell proliferation.

Vuk-Pavlovic, Z.; Rohrbach, M.S.



Lymphocyte and monocyte flow cytometry immunophenotyping as a diagnostic tool in uncharacteristic inflammatory disorders  

PubMed Central

Background Patients with uncharacteristic inflammatory symptoms such as long-standing fatigue or pain, or a prolonged fever, constitute a diagnostic and therapeutic challenge. The aim of the present study was to determine if an extended immunophenotyping of lymphocytes and monocytes including activation markers can define disease-specific patterns, and thus provide valuable diagnostic information for these patients. Methods Whole blood from patients with gram-negative bacteraemia, neuroborreliosis, tuberculosis, acute mononucleosis, influenza or a mixed connective tissue disorders, as diagnosed by routine culture and serology techniques was analysed for lymphocyte and monocyte cell surface markers using a no-wash, no-lyse protocol for multi-colour flow cytometry method. The immunophenotyping included the activation markers HLA-DR and CD40. Plasma levels of soluble TNF alpha receptors were analysed by ELISA. Results An informative pattern was obtained by combining two of the analysed parameters: (i), the fractions of HLA-DR-expressing CD4+ T cells and CD8+ T cells, respectively, and (ii), the level of CD40 on CD14+ CD16- monocytes. Patients infected with gram-negative bacteria or EBV showed a marked increase in monocyte CD40, while this effect was less pronounced for tuberculosis, borrelia and influenza. The bacterial agents could be distinguished from the viral agents by the T cell result; CD4+ T cells reacting in bacterial infection, and the CD8+ T cells dominating for the viruses. Patients with mixed connective tissue disorders also showed increased activation, but with similar engagement of CD4+ and CD8+ T cells. Analysis of soluble TNF alpha receptors was less informative due to a large inter-individual variation. Conclusion Immunophenotyping including the combination of the fractions of HLA-DR expressing T cell subpopulations with the level of CD40 on monocytes produces an informative pattern, differentiating between infections of bacterial and viral origin. Furthermore, a quantitative analysis of these parameters revealed the novel finding of characteristic patterns indicating a subacute bacterial infection, such as borreliosis or tuberculosis, or a mixed connective tissue disorder. The employed flow cytometric method is suitable for clinical diagnostic laboratories, and may help in the assessment of patients with uncharacteristic inflammatory symptoms.



HLA-DR human histocompatibility leukocyte antigens-restricted lymphocyte-monocyte interactions in the release from monocytes of acidic isoferritins that suppress hematopoietic progenitor cells.  

PubMed Central

Acidic isoferritins, which under normal conditions are released from monocytes and macrophages, have a suppressive effect in vitro on granulocyte-macrophage, erythroid, and multipotential hematopoietic progenitor cells. Cell interactions modulating the release of acidic isoferritin-inhibitory activity (AIFIA) from human monocytes were investigated using the bone marrow granulocyte-macrophage progenitor cells as a target cell assay for assessing AIFIA. Monocytes, in the absence of T lymphocytes, released AIFIA when allowed to condition culture medium at 10(4) or higher concentrations of monocytes/ml. However, subpopulations of T lymphocytes modulated the release of AIFIA from monocytes. OKT8+- and OKT4+-T lymphocytes were obtained from E-rosette-positive lymphocytes by using T lymphocyte subset-specific monoclonal antibodies in either a complement-dependent cytotoxicity test to select negatively for the cells or by selection using a "panning" procedure. OKT8+-T lymphocytes suppressed completely and OKT4+-T lymphocytes enhanced the constitutive release of AIFIA from monocytes. OKT4+ lymphocytes also induced the release of AIFIA from concentrations of 10(3) monocytes/ml which did not release measurable amounts of AIFIA by themselves. The release of AIFIA from monocytes involved HLA-DR+-monocytes and -T lymphocytes. Pulsing monocytes with monoclonal antibodies to framework determinants on HLA-DR molecules, in the absence of complement, did not influence the constitutive release of AIFIA. Pulsing monocytes or T lymphocyte subpopulations with such antibodies, in the absence of complement, blocked the suppressing and inducing activities of the appropriate subpopulations of T lymphocytes. Monoclonal antibodies to common determinants shared by HLA-A, B, and C molecules did not block these cellular interactions. Treating monocytes and T lymphocytes in a complement-dependent cytotoxicity test with dilutions of the anti-HLA-DR antibodies that did not block the cellular interactions removed the populations of monocytes constitutively releasing AIFIA and the T lymphocyte subsets modulating this release. Modulation of the release of AIFIA from monocytes by T lymphocyte subpopulations required the use of autologous cells, cells from HLA-identical siblings, or unrelated donors matched for HLA-DR. Matching for only one HLA haplotype gave partial responses and this was seen in testing cells from related individuals as well as among unrelated test combinations. These cellular interactions were not detected with HLA-DR-incompatible cells differing for two HLA-DR antigens. Admixture of such HLA-DR- incompatible allogeneic cells did not interfere with the regulation of AIFIA release in the autologous cell interactions. Thus, release of AIFIA from monocytes is restricted genetically by HLA-DR at the level of T lymphocyte-monocyte interactions. The genetic determinants on the HLA-class II molecules that induce stimulation in vitro in mixed lymphocyte culture (i.e., HLA-D), however, were not involved in this effort.

Broxmeyer, H E; Juliano, L; Lu, L; Platzer, E; Dupont, B




Microsoft Academic Search

TNF-? and IL-1 were reported to be the most powerful inducers of IL-8 in a multitude of cells, including leukocytes. In this study, we investigated TNF-?- and IL-1-mediated regulation of IL-8 gene expression in non-fractionated PBMC, and purified monocyte (MO) and lymphocyte (LY) fractions. Our analysis revealed that purified human MO did not respond to exogenous TNF-? with the induction

Yuri V Chaly; Rathinam S Selvan; Konstantin V Fegeding; Tatyana S Kolesnikova; Nikolai N Voitenok



Isolation of human and porcine monocytes and lymphocytes by three parameter flow sorting.  


Monocytes and lymphocytes were separated from total human blood cells by Ficoll-paque gradient centrifugation (at 600 g) followed by two parameter fluorescence activated cell sorting (forward [FLS] and perpendicular light scatter [PLS]). For human blood cells this technique gives good separation and high purity of the monocytes (more than 85%). For porcine blood cells we modified the Ficoll-paque gradient centrifugation-step by centrifuging at a lower speed (250 g). Since two parameter flow cytometry of porcine leukocytes (FLS and PLS) gave poor resolution we added endogenous non-specific esterase activity as a third parameter using fluorescein diacetate (FDA) as a fluorogenic substrate. The sorted fractions were cytocentrifuged and purity was checked with hematoxylin and/or peroxidase staining. Moreover, monoclonal antibodies against monocyte cell surface antigens were used to evaluate the purity of the sorted fractions. Three parameter sorting (PLS, FLS and fluorescence) yields good purification of porcine monocytes (86 +/- 1% pure) and lymphocytes (81 +/- 2% pure). There was a substantial adhesion of porcine monocytes (326 +/- 25/mm2) and lymphocytes (146 +/- 21/mm2) to monolayers of porcine aorta endothelial cells (PAEC). PMID:2123590

de Gruijter, M; van Rijn, M A; Verkerk, A; Jongkind, J F



Maternal microchimerism in healthy adults in lymphocytes, monocyte\\/macrophages and NK cells  

Microsoft Academic Search

During pregnancy some maternal cells reach the fetal circulation. Microchimerism (Mc) refers to low levels of genetically disparate cells or DNA. Maternal Mc has recently been found in the peripheral blood of healthy adults. We asked whether healthy women have maternal Mc in T and B lymphocytes, monocyte\\/macrophages and NK cells and, if so, at what levels. Cellular subsets were

Laurence S Loubière; Nathalie C Lambert; Laura J Flinn; Timothy D Erickson; Zhen Yan; Katherine A Guthrie; Kathy T Vickers; J Lee Nelson



Serine leucocyte proteinase inhibitor-treated monocyte inhibits human CD4+ lymphocyte proliferation  

PubMed Central

Serine leucocyte proteinase inhibitor (SLPI) is the main serine proteinase inhibitor produced by epithelial cells and has been shown to be a pleiotropic molecule with anti-inflammatory and microbicidal activities. However, the role of SLPI on the adaptive immune response is not well established. Therefore, we evaluated the effect of SLPI on lymphocyte proliferation and cytokine production. Human peripheral blood mononuclear cells (PBMC) were treated with mitogens plus SLPI and proliferation was assessed by [3H]thymidine uptake. The SLPI decreased the lymphocyte proliferation induced by interleukin-2 (IL-2) or OKT3 monoclonal antibodies in a dose-dependent manner. Inhibition was not observed when depleting monocytes from the PBMC and it was restored by adding monocytes and SLPI. SLPI-treated monocyte slightly decreased MHC II and increased CD18 expression, and secreted greater amounts of IL-4, IL-6 and IL-10 in the cell culture supernatants. SLPI-treated monocyte culture supernatant inhibited the CD4+ lymphocyte proliferation but did not affect the proliferation of CD8+ cells. Moreover, IL-2 increased T-bet expression and the presence of SLPI significantly decreased it. Finally, SLPI-treated monocyte culture supernatant dramatically decreased interferon-? but increased IL-4, IL-6 and IL-10 in the presence of IL-2-treated T cells. Our results demonstrate that SLPI target monocytes, which in turn inhibit CD4 lymphocyte proliferation and T helper type 1 cytokine secretion. Overall, these results suggest that SLPI is an alarm protein that modulates not only the innate immune response but also the adaptive immune response.

Guerrieri, Diego; Tateosian, Nancy L; Maffia, Paulo C; Reiteri, Romina M; Amiano, Nicolas O; Costa, Maria J; Villalonga, Ximena; Sanchez, Mercedes L; Estein, Silvia M; Garcia, Veronica E; Sallenave, Jean-Michel; Chuluyan, Hector E



Serine leucocyte proteinase inhibitor-treated monocyte inhibits human CD4(+) lymphocyte proliferation.  


Serine leucocyte proteinase inhibitor (SLPI) is the main serine proteinase inhibitor produced by epithelial cells and has been shown to be a pleiotropic molecule with anti-inflammatory and microbicidal activities. However, the role of SLPI on the adaptive immune response is not well established. Therefore, we evaluated the effect of SLPI on lymphocyte proliferation and cytokine production. Human peripheral blood mononuclear cells (PBMC) were treated with mitogens plus SLPI and proliferation was assessed by [(3) H]thymidine uptake. The SLPI decreased the lymphocyte proliferation induced by interleukin-2 (IL-2) or OKT3 monoclonal antibodies in a dose-dependent manner. Inhibition was not observed when depleting monocytes from the PBMC and it was restored by adding monocytes and SLPI. SLPI-treated monocyte slightly decreased MHC II and increased CD18 expression, and secreted greater amounts of IL-4, IL-6 and IL-10 in the cell culture supernatants. SLPI-treated monocyte culture supernatant inhibited the CD4(+) lymphocyte proliferation but did not affect the proliferation of CD8(+) cells. Moreover, IL-2 increased T-bet expression and the presence of SLPI significantly decreased it. Finally, SLPI-treated monocyte culture supernatant dramatically decreased interferon-? but increased IL-4, IL-6 and IL-10 in the presence of IL-2-treated T cells. Our results demonstrate that SLPI target monocytes, which in turn inhibit CD4 lymphocyte proliferation and T helper type 1 cytokine secretion. Overall, these results suggest that SLPI is an alarm protein that modulates not only the innate immune response but also the adaptive immune response. PMID:21574992

Guerrieri, Diego; Tateosian, Nancy L; Maffía, Paulo C; Reiteri, Romina M; Amiano, Nicolás O; Costa, María J; Villalonga, Ximena; Sanchez, Mercedes L; Estein, Silvia M; Garcia, Verónica E; Sallenave, Jean-Michel; Chuluyan, Héctor E



Monocyte-dependent death of freshly isolated T lymphocytes: induction by phorbolester and mitogens and differential effects of catalase.  


Resting T cells are resistant to anti-Fas (CD95) mAb-mediated apoptosis but undergo apoptosis when triggered by anti-CD3 mAb or phorbolester PMA in the presence of PMA-activated monocytes. In this study, PMA, as well as the mitogens PHA and Con A, was found to induce death of resting T cells in the presence of autologous or allogeneic monocytes, while PWM was ineffective. Although several established monocytic and myelocytic cell lines were potent accessory cells for the mitogen-induced expansion of T lymphocytes, they all failed to replace plastic-adherent monocytes in the induction of monocyte-dependent cell death (MDCD) by PMA or PHA. CD45RA-positive cord blood T cells were as susceptible as peripheral blood T cells from adult donors to PMA-stimulated induction of MDCD. Using optimal concentrations of phorbolester, MDCD was inhibited neither by Fas-Fc fusion protein or neutralizing anti-Fas mAb, nor by inhibitors of IL-1beta-converting enzyme (ICE)-like proteases. In striking contrast, the H2O2 scavenger catalase completely prevented the PMA-stimulated T cell death, thereby revealing a potent mitogenic activity of PMA for human T cells in the presence of monocytes. Taken together, our results demonstrate that the accessory cell activity of monocytes/macrophages can be separated into "T cell death" and "T cell expansion" costimulatory functions, of which only the latter is mediated by established cell lines. Moreover, our results point to a pivotal role of reactive oxygen intermediates in the execution of MDCD triggered by PMA. PMID:9686585

Wesch, D; Marx, S; Kabelitz, D



Immature Dendritic Cells Generated from Cryopreserved Human Monocytes Show Impaired Ability to Respond to LPS and to Induce Allogeneic Lymphocyte Proliferation  

PubMed Central

Dendritic cells play a key role in the immune system, in the sensing of foreign antigens and triggering of an adaptive immune response. Cryopreservation of human monocytes was investigated to understand its effect on differentiation into immature monocyte-derived dendritic cells (imdDCs), the response to inflammatory stimuli and the ability to induce allogeneic lymphocyte proliferation. Cryopreserved (crp)-monocytes were able to differentiate into imdDCs, albeit to a lesser extent than freshly (frh)-obtained monocytes. Furthermore, crp-imdDCs had lower rates of maturation and cytokine/chemokine secretion in response to LPS than frh-imdDCs. Lower expression of Toll-like receptor 4 (at 24 and 48 h) and higher susceptibility to apoptosis in crp-imdDCs than in fresh cells would account for the impaired maturation and cytokine/chemokine secretion observed. A mixed leukocyte reaction showed that lymphocyte proliferation was lower with crp-imdDCs than with frh-imdDCs. These findings suggested that the source of monocytes used to generate human imdDCs could influence the accuracy of results observed in studies of the immune response to pathogens, lymphocyte activation, vaccination and antigen sensing. It is not always possible to work with freshly isolated monocytes but the possible effects of freezing/thawing on the biology and responsiveness of imdDCs should be taken into account.

Silveira, Guilherme Ferreira; Wowk, Pryscilla Fanini; Machado, Analia Maria Breckenfeld; dos Santos, Claudia Nunes Duarte; Bordignon, Juliano



Regulation of B7-H1 expression on peripheral monocytes and IFN-? secretion in T lymphocytes by HBeAg.  


This study is to observe the expression of B7-H1, PD-1 and TLR2 on peripheral blood monocytes (PBMCs) regulated by HBeAg in chronic hepatitis B (CHB), and to illustrate the relation between HBeAg and persistent infection of HBV. In both CHB patients and healthy controls, the expression of B7-H7 was significantly increased on CD14(+) monocytes incubated with HBeAg, while that of TLR2 was significantly reduced; the expression of specific IFN-? was significantly decreased in CD3(+)CD4(+) T lymphocytes incubated with HBeAg, while IL-6 and IL-10 in conditioned media were significantly increased. HBeAg is able to significantly up-regulate B7-H1, down-regulate TLR2 on monocytes, reduce IFN-? produced by T lymphocytes and increase Th2-type cytokines secretion. These findings suggest that HBeAg suppresses the specific cellular immunity to clear the virus, and eventually lead to immune tolerance to HBV infection. Therefore, HBeAg plays an important role in immune suppression in chronic HBV patients. PMID:23850673

Han, Yaping; Li, Jun; Jiang, Longfeng; Xu, Qingqing; Liu, Bo; Jin, Ke; Liu, Yuan; Huang, Zuhu



Diallyl disulfide reduced dose-dependently the number of lymphocyte subsets and monocytes in rats.  


Diallyl disulfide (DADS) is a major sulfur compound of garlic, and exerts anti-inflammatory, immune-modulatory, and enhancing sympathetic activity effects. However, it still remains unclear how DADS affects the distribution of white blood cell subsets, which is essential to execute effective immune responses and partially regulated by adrenal glucocorticoids. Therefore, we examined the dose-dependent effects of DADS administration on the circulating number of white blood cells (WBCs) and lymphocyte subsets, and plasma corticosterone concentration in rats. Male 10-wk-old Sprague Dawley rats were divided into the DADS-free and DADS-orally administered (dose=10, 20, and 40 mg/kg BW) groups. Blood samples were collected from the tail vein at 0, 1, 2, 4, and 6 h after the administration. DADS administration decreased dose- and time-dependently the circulating number of total WBCs, total lymphocytes, and monocytes. Within the lymphocyte subsets, the circulating number of T-lymphocytes and B-lymphocytes was significantly reduced 4 h after DADS administration in a dose-dependent manner, although that of natural killer (NK) cells was not affected. On the other hand, although DADS administration did not significantly change the circulating number of neutrophils, the circulating number of eosinophils and basophils showed a decreasing tendency after DADS administration. In contrast, plasma corticosterone concentration was increased 2 h after DADS administration in a dose-dependent manner. These results suggest that DADS administration reduces the circulating number of monocytes and lymphocytes, including especially acquired immune cells, via the action of corticosterone, and the effects are induced in a dose-dependent manner. PMID:23132314

Hashizume, Yoko; Shirato, Ken; Abe, Ikumi; Kobayashi, Ayumu; Mitsuhashi, Ryosuke; Shiono, Chikako; Sato, Shogo; Tachiyashiki, Kaoru; Imaizumi, Kazuhiko



Effect of vitamin E modified cellulose membrane on human lymphocyte, monocyte, and granulocyte CD11b/CD18 adhesion molecule expression during hemodialysis.  


Chronic renal failure induces a clinical state of immunodefi ciency that also depends upon a wide spectrum of dialysis membranes used during hemodialysis. Previous studies have shown that cellular immunodeficiency is caused by malfunc tion of the antigen presenting cells (monocytes or granulocytes). Subsequent activation of rolling mononuclear leuko cytes results in up-regulated expression of CD11b/CD18 (Mac-1) on endothelial cells. It is postulated that a VitE coated dialysis membrane might minimize the membrane biocompatibility, thereby generating a smaller amount of re active oxygen species (ROS). The purpose of this study was to evaluate the expression of the CD11b/CD18 adhesion mole cule on lymphocytes, monocytes, and granulocytes during HD in 10 patients, using flow cytometric analysis. The study protocol included the measurement of molecule expression using cellulose membrane (Clirans RS15, TERUMO Corp. Japan), and the same membrane coated by vitamin E (Exce brane, Clirans E15, TERUMO Corp., Japan) during 20 dialysi sessions each. Lymphocyte CD11 b/CD1 8 (Mac-1) expression did not change with either dialyzer type. However, monocyt (p = 0.046) and granulocyte (p = 0.018) CD11b/CD18 ex pression in the post HD period was significantly lower using the vitamin E coated membrane compared with the contro cellulose membrane. Our findings suggest a significant de crease in activation and migration of monocytes and granu locytes when using a vitamin E coated cellulose membrane. PMID:11730199

Zaluska, W T; Ksiazek, A; Rolíski, J


ROS release and Hsp70 expression after exposure to 1,800 MHz radiofrequency electromagnetic fields in primary human monocytes and lymphocytes  

Microsoft Academic Search

The aim of this study is to investigate if 1,800 MHz radiofrequency electromagnetic fields (RF-EMF) can induce reactive oxygen species (ROS) release and\\/or changes in heat shock protein 70 (Hsp70) expression in human blood cells, using different exposure and co-exposure conditions. Human umbilical cord blood-derived monocytes and lymphocytes were used to examine ROS release after exposure to continuous wave or different

M. Lantow; M. Lupke; J. Frahm; M. O. Mattsson; N. Kuster; M. Simko



Inflammatory monocytes activate memory CD8+ T and innate NK lymphocytes independent of cognate antigen during microbial pathogen invasion  

PubMed Central

SUMMARY Memory CD8+ T cells induced upon immunization exhibit improved functional features that contribute to protection of immunized hosts. Although both cognate antigen recognition and inflammation are important for memory CD8+ T cell reactivation, the relative contribution of these factors and the cell types providing these signals in vivo are poorly defined. Here, we show that Ly6C+CCR2+ inflammatory monocytes, a subset of monocytes, largely orchestrate memory CD8+ T and NK lymphocytes activation by differentiating into interleukin-18 (IL-18)- and IL-15-producing cells in an inflammasome and type I interferon-IRF3-dependent manner. Memory CD8+ T cells became potent effector cells by sensing inflammation from monocytes independently of their cognate antigen. Like NK cells, they underwent rapid mobilization, upregulated intense and sustained effector functions during bacterial, viral and parasitic infections, and contributed to innate responses and protection in vivo. Thus, inflammatory monocyte-derived IL-18 and IL-15 are critical to initiate memory CD8+ T and NK lymphocytes differentiation into antimicrobial effector cells.

Soudja, Saidi M'Homa; Ruiz, Anne L.; Marie, Julien C.; Lauvau, Gregoire



Leukoreduction system chambers are an efficient, valid, and economic source of functional monocyte-derived dendritic cells and lymphocytes.  


The demand for human monocyte-derived dendritic cells (moDCs), as well as for primary human B and T lymphocytes for immunological research purposes has been increased in recent years. Classically, these monocytes are isolated from blood, leukapheresis products or buffy coats of healthy donors by plastic adherence of peripheral blood mononuclear cells (PBMCs), followed by stimulation with granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4, while lymphocytes are usually isolated from the non-adherent fraction (NAF) by magnetic cell sorting. However, donor-blood is a limited resource and not every blood bank offers leukapheresis products or buffy coats for laboratory use. Additionally, a leukapheresis is very expensive and also the generation/isolation of cells is time- and cost-intensive. To overcome some of these obstacles, we evaluated if low-cost leukoreduction system chambers (LRSCs), which arise after routine donor plateletpheresis procedures, and are usually discarded, would be an alternative and appropriate source of PBMCs to generate moDCs and to isolate lymphocytes. By analyzing the number and phenotype of immature and mature dendritic cells (DCs), as well as of B and T lymphocytes derived from LRSCs, we found all cells to be of high quantity and quality. Further investigations on DCs comprising transwell migration assays, allogeneic mixed lymphocyte reactions (MLR), cytokine secretion assays, and cytotoxic T cell induction assays revealed high migratory, as well as stimulatory capacity of these cells. In addition, DCs and T cells were efficiently electroporated with mRNA and showed characteristic cytokine production after co-culture, demonstrating LRSCs as an efficient, valid, and economic source for generation of moDCs and lymphocytes for research purposes. PMID:23932569

Pfeiffer, Isabell A; Zinser, Elisabeth; Strasser, Erwin; Stein, Marcello F; Dörrie, Jan; Schaft, Niels; Steinkasserer, Alexander; Knippertz, Ilka



ROS release and Hsp70 expression after exposure to 1,800 MHz radiofrequency electromagnetic fields in primary human monocytes and lymphocytes.  


The aim of this study is to investigate if 1,800 MHz radiofrequency electromagnetic fields (RF-EMF) can induce reactive oxygen species (ROS) release and/or changes in heat shock protein 70 (Hsp70) expression in human blood cells, using different exposure and co-exposure conditions. Human umbilical cord blood-derived monocytes and lymphocytes were used to examine ROS release after exposure to continuous wave or different GSM signals (GSM-DTX and GSM-Talk) at 2 W/kg for 30 or 45 min of continuous or intermittent (5 min ON/5 min OFF) exposure. The cells were exposed to incubator conditions, to sham, to RF-EMF, or to chemicals in parallel. Cell stimulation with the phorbol ester phorbol-12-myristate-13-acetate (PMA; 1 microM) was used as positive control for ROS release. To investigate the effects on Hsp70 expression, the human monocytes were exposed to the GSM-DTX signal at 2 W/kg for 45 min, or to heat treatment (42 degrees C) as positive control. ROS production and Hsp70 expression were determined by flow cytometric analysis. The data were compared to sham and/or to control values and the statistical analysis was performed by the Student's t-test (P<0.05). The PMA treatment induced a significant increase in ROS production in human monocytes and lymphocytes when the data were compared to sham or to incubator controls. After continuous or intermittent GSM-DTX signal exposure (2 W/kg), a significantly different ROS production was detected in human monocytes if the data were compared to sham. However, this significant difference appeared due to the lowered value of ROS release during sham exposure. In human lymphocytes, no differences could be detected if data were compared either to sham or to incubator control. The Hsp70 expression level after 0, 1, and 2 h post-exposure to GSM-DTX signal at 2 W/kg for 1 h did not show any differences compared to the incubator or to sham control. PMID:16552570

Lantow, M; Lupke, M; Frahm, J; Mattsson, M O; Kuster, N; Simko, M



Intracellular cytokine profile of cord and adult blood monocytes  

Microsoft Academic Search

Cord blood (CB) transplantations are associated with low graft-versus-host disease (GVHD). The pathophysiology of GVHD involves interaction and activation of different cell types, as lymphocytes and monocytes, and results in a cascade of cytokine production. After antigen or mitogen stimulation, CB monocytes release lower levels of cytokines than adult blood (AB) monocytes. In this study, the detection of intracellular IL-1?

B Brichard; I Varis; D Latinne; V Deneys; M de Bruyere; P Leveugle; G Cornu



Histamine inhibits adhesion molecule expression in human monocytes, induced by advanced glycation end products, during the mixed lymphocyte reaction  

PubMed Central

Background and purpose: Post-transplant diabetes mellitus is a frequent complication among transplant recipients. Ligation of advanced glycation end products (AGEs) with their receptor on monocytes/macrophages plays important roles in the genesis of diabetic complications. The enhancement of adhesion molecule expression on monocytes/macrophages activates T-cells, reducing allograft survival. Out of four distinct AGE subtypes (AGE-2, AGE-3, AGE-4 and AGE-5), only AGE-2 and AGE-3 induced expression of intercellular adhesion molecules (ICAMs), output of cytokines and proliferation of lymphocytes, during the mixed lymphocyte reaction (MLR). Here we have assessed the role of histamine in the actions of AGEs during the MLR. Experimental approach: Human peripheral blood cells were used in these experiments. Flow cytometry was used to examine the expression of the ICAM-1, B7.1, B7.2 and CD40. Production of the cytokine interferon-?, and levels of cAMP were determined by elisa. Lymphocyte proliferation was determined by [3H]-thymidine uptake. Key results: Histamine concentration dependently inhibited the action of AGE-2 and AGE-3. The actions of histamine were antagonized by an H2-receptor antagonist, famotidine, and mimicked by H2/H4-receptor agonists, dimaprit and 4-methylhistamine. The effects of histamine were reversed by a protein kinase A (PKA) inhibitor, H89, and mimicked by dibutyryl cAMP and an adenylate cyclase activator, forskolin. Conclusions and implications: Histamine down-regulated AGE-2- and AGE-3-induced expression of adhesion molecules, cytokine production and lymphocyte proliferation via histamine H2 receptors and the cAMP/PKA pathway.

Zhang, J; Takahashi, HK; Liu, K; Wake, H; Liu, R; Sadamori, H; Matsuda, H; Yagi, T; Yoshino, T; Mori, S; Nishibori, M



Immunogenetic analysis of cellular interactions governing the recruitment of T lymphocytes and monocytes in lymphocytic choriomeningitis virus-induced immunopathology  

Microsoft Academic Search

The Lyt2+ class I major histocompatibility complex (MHC)-restricted virus-immune T cells that induce murine lymphocytic choriomeningitis (LCM) are targeted onto radiation-resistant cells in the central nervous system of virus-infected mice. The use of appropriate bone marrow radiation chimeras as LCM virus-infected, (immunosuppressed recipients for immune T-cell transfer has established that, though bone marrow-derived cells can stimulate virus-specific cytotoxic T lymphocytes

Peter C. Doherty; Rhodri Ceredig; Jane E. Allan



Ceramide stimulates the uptake of neutral red in human neutrophils, monocytes, and lymphocytes  

Microsoft Academic Search

Neutral red is a vital stain known to be accumulated in the lysosomes of neutrophils and monocytes. It is used mainly to\\u000a identify and detect the activated state of these cells. We have found that the extracellular application of physiological\\u000a ceramide, i.e., a product of sphingomyelin hydrolysis and a newly defined intracellular second-messenger substance, increased\\u000a the uptake of neutral red

S. Sipka; P. Antal-Szalmás; I. Szöllösi; I. Csipö; G. Lakos; G. Szegedi



Soluble ions more than particulate cobalt-alloy implant debris induce monocyte costimulatory molecule expression and release of proinflammatory cytokines critical to metal-induced lymphocyte reactivity.  


Aseptic osteolysis has been associated with excessive immune reactivity to particulate implant debris; however, innate and adaptive immune mechanisms that underlie implant debris reactivity remain incompletely understood. Although particulate debris has been implicated as the major type of implant debris mediating macrophage-induced osteolysis, the degree to which metal ions affect a proinflammatory response (if at all) remains unknown. We hypothesized that both soluble and particulate metal implant debris will induce proinflammatory responses in human monocytes resulting in cytokine production and elevated expression of T cell costimulatory molecules, facilitating adaptive immune responses. We tested this hypothesis by characterizing the response of a human monocyte cell line (THP-1), isolated primary human monocytes and PBMCs challenged with Co-Cr-Mo alloy particles and soluble cobalt, chromium, molybdenum, and nickel ions. Our results indicate that soluble cobalt, nickel, and molybdenum can induce monocyte up-regulation of T cell costimulatory molecules (CD80, CD86, ICAM-1) in human monocytes/macrophages. Furthermore, cobalt, molybdenum ions, and Co-Cr-Mo alloy particles similarly induce elevated secretion of IL-1beta, TNFalpha, and IL-6. Antibody blockade of CD80 and CD86, crucial secondary molecules for adaptive responses, abrogated lymphocyte reactivity to metal challenge in metal reactive subjects. Also the addition of IL-1 receptor antagonist (IL-1ra), (which indirectly blocks pro-IL-1beta and thus IL-1beta release), significantly reduced lymphocyte reactivity in metal-reactive subjects. Thus, both soluble and particulate metal implant debris induce monocyte/macrophage proinflammatory responses that are metal and individual specific. This suggests metal-induced up-regulation of costimulatory molecules and proinflammatory cytokine production is necessary to induce lymphocyte activation/proliferation to metal implant debris. PMID:19844976

Caicedo, Marco S; Pennekamp, Peter H; McAllister, Kyron; Jacobs, Joshua J; Hallab, Nadim J



Respective roles and interactions of T-lymphocyte and PGE2-mediated monocyte suppressive activities in human newborns and mothers at the time of delivery  

SciTech Connect

Recently the concept of a poorly functional humoral immune response in the newborn was proposed. Data have been presented indicating that the impaired newborn B cell maturation, as shown in vitro in a pokeweed mitogen-induced B cell maturation system, is due both to an immaturity of lymphocyte subsets and to an increased suppressive T activity. In the present work, we present evidence that there exists a predominance of a naturally occurring T lymphocyte suppressive activity in the cord blood in that the removal of the suppressive activity by irradiation allows a normal maturation of newborn B cells. Such normal maturation of newborn B cells can also be obtained using mixed cultures of adult T cells and newborn B cells. Newborn suppressor T cells belong to both EA gamma (+) and EA gamma (-) fractions, and it is not known whether these two groups do or do not belong to different subsets. The PGE2-dependent monocyte suppressive activity does not play any role in the suppression observed in newborns since newborn monocytes are poorly suppressive and since they produce a smaller amount of PGE2 than adult monocytes. Some observations suggest, on the contrary, that the suppressive T lymphocytes can regulate the level of the PGE2-dependent monocyte suppressive activity. It should be noticed that similar observations about T lymphocyte and PGE2-dependent monocyte suppressive activities have been made at the same time using mothers' cells. These observations suggest the possibility that such changes in B cell immune regulation may result from an interaction between maternal and fetal lymphoid cells.

Durandy, A.; Fischer, A.; Mamas, S.; Dray, F.; Griscelli, C.



Immunologic effector mechanisms of a standardized mistletoe extract on the function of human monocytes and lymphocytes in vitro, ex vivo, and in vivo.  


Even though mistletoe extracts have been in clinical use for centuries their exact mode of action is still unknown. Currently, the application scheme for registered preparations is a dose-escalating scheme to thus reduce side effects. In this study, healthy controls and patients were evaluated for their immunologic response to treatment with a standardized mistletoe extract (Iscador). It shows a strong effect as adjuvant that induces TNF-alpha and IL-12, which was partly mediated via CD14. Desensitization of the TNF-alpha response could be shown after repeated application in vitro and in vivo. Furthermore, Iscador induces a specific lymphocyte sensitization upon multiple injections and production of IgG1- and IgG3 -mistletoe antibodies. Remarkably, a systemic bystander effect (heterologous immunity against other recall antigens) was observed after long-term treatment. In conclusion, dose-escalation reduces the monocyte-related clinical side effects. A T-lymphocyte sensitization stimulates mainly a specific Th1 response. The most interesting clinical long-term effect is the bystander stimulation of various memory T cells that might mediate in vivo antitumor and antiinfectious T-cell response under mistletoe-extract immunization. PMID:16705487

Heinzerling, Lucie; von Baehr, Volker; Liebenthal, Christa; von Baehr, Rüdiger; Volk, Hans-Dieter



Induction of tumor necrosis factor and interleukin-1 by purified staphylococcal toxic shock syndrome toxin 1 requires the presence of both monocytes and T lymphocytes.  

PubMed Central

Highly purified staphylococcal toxic shock syndrome toxin 1 (TSST-1) was tested for its ability to induce the cytokines tumor necrosis factor (TNF) and interleukin-1 (IL-1) from fractionated human peripheral blood mononuclear cells prepared from seven healthy donors. Highly purified monocytes alone or T lymphocytes alone did not produce TNF or IL-1 when incubated with TSST-1 at 37 degrees C for up to 72 h. However, the addition of 10 micrograms of TSST-1 per ml to a 1:1 mixture of monocytes and T cells resulted in significant TNF (predominantly TNF-alpha) and IL-1 beta production after 24 h at 37 degrees C. The nature of the monocyte/T-cell interaction did not appear to involve gamma interferon (IFN-gamma), since 10 micrograms of rabbit anti-IFN-gamma per ml did not neutralize TNF-alpha production after TSST-1 induction. Similarly, L243, a monoclonal antibody to HLA-DR which blocks TSST-1 binding to monocytes, did not inhibit TNF-alpha production following TSST-1 induction. However, direct contact between monocytes and T cells was required, since physical separation of cells in double-chamber culture wells abolished TNF-alpha secretion after TSST-1 stimulation. Furthermore, paraformaldehyde fixation of either monocytes or T cells prior to addition to viable T cells or monocytes, respectively, also abolished TNF-alpha secretion, suggesting that aside from cell contact, soluble factors were also involved. Our results suggest that cytokine production involves more than binding of TSST-1 to its receptor on monocytes alone and that cell contact with T cells and the release of a soluble factor(s) other than IFN-gamma may be essential for the induction of cytokines by this toxin.

See, R H; Kum, W W; Chang, A H; Goh, S H; Chow, A W



Virus production and viral antigen expression in porcine blood monocytes inoculated with pseudorabies virus  

Microsoft Academic Search

Summary Interactions of pseudorabies virus (PRV) with peripheral blood mononuclear cells (PBMC) were studied. T-lymphocytes, B-lymphocytes and monocytes were selected or depleted from the PBMC fraction by means of several separation techniques. After inoculation with virulent PRV in vitro, the percentage of cells that expressed viral antigens was determined in the different subpopulations by immunofluorescence. The susceptibility of monocytes depended

H. J. Nauwynck; M. B. Pensaert



Identification of alpha-naphthyl butyrate esterase as a plasma membrane ectoenzyme of monocytes and as a discrete intracellular membrane- bounded organelle in lymphocytes  

PubMed Central

A reaction for an esterase, with a nonhalogenated, short-chain naphthyl ester (alpha-naphthyl butyrate or alpha-naphthyl acetate) as the substrate, has been used to identify mononuclear phagocytes by light microscopy. By analyzing techniques used in the collection, separation, fixation, processing, and embedding of human blood leukocytes for electron microscopy, we adapted the light microscopic method for use in determining the fine structural localization of this reaction. In monocytes, the reaction product covered the external surface of the plasma membrane. This distribution indicated that monocytic esterase is an ectoenzyme. The addition of NaF completely inhibited the monocytic reaction. In lymphocytes, the reaction product was localized in membrane-bounded intracellular organelles, similar to those previously shown to contain phospholipid and called Gall bodies. These organelles correspond with the punctate densities or focal reaction product observed by light microscopy. Other investigators believe that this distribution of enzyme in lymphocytes marks a subset of T cells, the Tmicro. The lymphocytic reaction was not inhibited by NaF.



Blood Lymphocyte-to-Monocyte Ratio Identifies High-Risk Patients in Diffuse Large B-Cell Lymphoma Treated with R-CHOP  

PubMed Central

Background Recent research has shown a correlation between immune microenvironment and lymphoma biology. This study aims to investigate the prognostic significance of the immunologically relevant lymphocyte-to-monocyte ratio (LMR), in diffuse large B-cell lymphoma (DLBCL) in the rituximab era. Methodology/Principal Findings We analyzed retrospective data from 438 newly diagnosed DLBCL patients treated with rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) therapy. We randomly selected 200 patients (training set) to generate a cutoff value for LMR by receiver operating characteristic (ROC) curve analysis. LMR was then analyzed in a testing set (n?=?238) and in all patients (n?=?438) for validation. The LMR cutoff value for survival analysis determined by ROC curve in the training set was 2.6. Patients with low LMR tended to have more adverse clinical characteristics. Low LMR at diagnosis was associated with worse survival in DLBCL, and could also identify high-risk patients in the low-risk IPI category. Multivariate analysis identified LMR as an independent prognostic factor of survival in the testing set and in all patients. Conclusions/Significance Baseline LMR, a surrogate biomarker of the immune microenvironment, is an effective prognostic factor in DLBCL patients treated with R-CHOP therapy. Future prospective studies are required to confirm our findings.

Xia, Yi; Sun, Jian; Huang, Ying; Wang, Yu; Zhu, Ying-Jie; Li, Ya-Jun; Zhao, Wei; Wei, Wen-Xiao; Lin, Tong-Yu; Huang, Hui-Qiang; Jiang, Wen-Qi



Immunization of Cattle by Infection with Cowdria ruminantium Elicits T Lymphocytes That Recognize Autologous, Infected Endothelial Cells and Monocytes  

Microsoft Academic Search

Peripheral blood mononuclear cells (PBMC) from immune cattle proliferate in the presence of autologous Cowdria ruminantium-infected endothelial cells and monocytes. Endothelial cells required treatment with T-cell growth factors to induce class II major histocompatibility complex expression prior to infection and use as stimulators. Proliferative responses to both infected autologous endothelial cells and monocytes were charac- terized by expansion of a



Student Research Award in the Undergraduate Degree Candidate category, 30th Annual Meeting of the Society for Biomaterials, Memphis, Tennessee, April 27-30, 2005. Monocyte/lymphocyte interactions and the foreign body response: in vitro effects of biomaterial surface chemistry.  


To determine the effect of biomaterial surface chemistry on leukocyte interaction and activity at the material/tissue interface, human peripheral blood monocytes and lymphocytes were cultured on a series of poly(ethylene terephthalate) (PET)-based biomaterials. Both monocytes and lymphocytes were isolated from whole human blood and separated by a nonadherent density centrifugation method before being plated on PET disks, surface modified by photograft copolymerization to yield hydrophobic, hydrophilic, anionic, and cationic surface properties. Monocytes and lymphocytes were cultured separately, to elicit baseline levels of activity, in direct coculture, to promote direct cell surface interactions, or in an indirect coculture system with both cell types separated by a -0.02-microm Transwell apparatus, to promote indirect paracrine interactions. Monocyte adhesion, macrophage fusion, and lymphocyte proliferation were measured on days 3, 7, 10, and 14 of culture. Results demonstrated that the presence of monocytes increased the activity of cocultured lymphocytes at the biomaterial/tissue interface, while the corresponding presence of lymphocytes increased the activation and fusion of indirectly cocultured monocytes. Biomaterial surface chemistry was also found to have a significant effect on monocyte adhesion and activation, and lymphocyte activity. Hydrophilic surfaces significantly inhibited both initial and longterm monocyte adhesion, and inhibited lymphocyte proliferation at longer time points. Anionic and cationic surfaces both exhibited mild inhibition of monocyte adhesion at prolonged time points and increased levels of macrophage fusion, while cationic surfaces decreased levels of lymphocyte proliferation and inhibited monocyte activity. These results elucidate the complex role of juxtacrine and paracrine interactions between monocytes and lymphocytes in the foreign body response, as well as promote the consideration of hydrophilic surfaces in future designs of implantable biomedical devices and prostheses. PMID:16124082

MacEwan, Matthew R; Brodbeck, William G; Matsuda, Takehisa; Anderson, James M



Antibody-dependent cell-mediated antibacterial activity of human mononuclear cells. I. K lymphocytes and monocytes are effective against meningococi in cooperation with human imune sera  

PubMed Central

In cooperation with human heat-inactivated antisera from adults immunized with group C meningococcal polysaccharide, normal human peripheral blood mononuclear cells significantly decreased the viability of group C meningococci (Mgc) in vitro. K lymphocytes (Null cells) and monocytes, (but not T or B lymphocytes) were capable of effecting antibody-dependent cell-mediated (ADC) antibacterial activity in this system. The degree to which meningococcal viability was decreased was a function of the length of the test incubation, the concentration of effector cells, and the amount of antiserum used in the assay. When specific antibodies directed against Mgc were adsorbed from the antiserum, cell-mediated antibacterial activity was abolished. ADC antibacterial activity was also abrogated by performing the assay at 4 degrees C or by heating effector cells to 46 degrees C for 15 min before the assay, Similarities between the ADC antibacterial system and previously described ADCC assays are discussed. The data suggest the K cells (as well as monocytes) may play a role in host immune defense against pathogenic bacteria.



Antibody-dependent cell-mediated antibacterial activity of human mononuclear cells. I. K lymphocytes and monocytes are effective against meningococi in cooperation with human imune sera.  


In cooperation with human heat-inactivated antisera from adults immunized with group C meningococcal polysaccharide, normal human peripheral blood mononuclear cells significantly decreased the viability of group C meningococci (Mgc) in vitro. K lymphocytes (Null cells) and monocytes, (but not T or B lymphocytes) were capable of effecting antibody-dependent cell-mediated (ADC) antibacterial activity in this system. The degree to which meningococcal viability was decreased was a function of the length of the test incubation, the concentration of effector cells, and the amount of antiserum used in the assay. When specific antibodies directed against Mgc were adsorbed from the antiserum, cell-mediated antibacterial activity was abolished. ADC antibacterial activity was also abrogated by performing the assay at 4 degrees C or by heating effector cells to 46 degrees C for 15 min before the assay, Similarities between the ADC antibacterial system and previously described ADCC assays are discussed. The data suggest the K cells (as well as monocytes) may play a role in host immune defense against pathogenic bacteria. PMID:109572

Lowell, G H; Smith, L F; Artenstein, M S; Nash, G S; MacDermott, R P



Generation of Transmitted/Founder HIV-1 Infectious Molecular Clones and Characterization of Their Replication Capacity in CD4 T Lymphocytes and Monocyte-Derived Macrophages  

PubMed Central

Genome sequences of transmitted/founder (T/F) HIV-1 have been inferred by analyzing single genome amplicons of acute infection plasma viral RNA in the context of a mathematical model of random virus evolution; however, few of these T/F sequences have been molecularly cloned and biologically characterized. Here, we describe the derivation and biological analysis of ten infectious molecular clones, each representing a T/F genome responsible for productive HIV-1 clade B clinical infection. Each of the T/F viruses primarily utilized the CCR5 coreceptor for entry and replicated efficiently in primary human CD4+ T lymphocytes. This result supports the conclusion that single genome amplification-derived sequences from acute infection allow for the inference of T/F viral genomes that are consistently replication competent. Studies with monocyte-derived macrophages (MDM) demonstrated various levels of replication among the T/F viruses. Although all T/F viruses replicated in MDM, the overall replication efficiency was significantly lower compared to prototypic “highly macrophage-tropic” virus strains. This phenotype was transferable by expressing the env genes in an isogenic proviral DNA backbone, indicating that T/F virus macrophage tropism mapped to Env. Furthermore, significantly higher concentrations of soluble CD4 were required to inhibit T/F virus infection compared to prototypic macrophage-tropic virus strains. Our findings suggest that the acquisition of clinical HIV-1 subtype B infection occurs by mucosal exposure to virus that is not highly macrophage tropic and that the generation and initial biological characterization of 10 clade B T/F infectious molecular clones provides new opportunities to probe virus-host interactions involved in HIV-1 transmission.

Ochsenbauer, Christina; Edmonds, Tara G.; Ding, Haitao; Keele, Brandon F.; Decker, Julie; Salazar, Maria G.; Salazar-Gonzalez, Jesus F.; Shattock, Robin; Haynes, Barton F.; Shaw, George M.; Hahn, Beatrice H.



Monocytic AML cells inactivate antileukemic lymphocytes: role of NADPH oxidase/gp91phox expression and the PARP-1/PAR pathway of apoptosis  

PubMed Central

Dysfunction of T cells and natural killer (NK) cells has been proposed to determine the course of disease in acute myeloid leukemia (AML), but only limited information is available on the mechanisms of lymphocyte inhibition. We aimed to evaluate to what extent human malignant AML cells use NADPH oxidase-derived reactive oxygen species (ROS) as an immune evasion strategy. We report that a subset of malignant myelomonocytic and monocytic AML cells (French-American-British [FAB] classes M4 and M5, respectively), recovered from blood or BM of untreated AML patients at diagnosis, expressed the NADPH oxidase component gp91phox. Highly purified FAB M4/M5 AML cells produced large amounts of ROS on activation and triggered poly-[ADP-ribose] polymerase-1?dependent apoptosis in adjacent NK cells, CD4+ T cells, and CD8+ T cells. In contrast, immature (FAB class M1) and myeloblastic (FAB class M2) AML cells rarely expressed gp91phox, did not produce ROS, and did not trigger NK or T-cell apoptosis. Microarray data from 207 AML patients confirmed a greater expression of gp91phox mRNA by FAB-M4/M5 AML cells than FAB-M1 cells (P < 10?11) or FAB-M2 cells (P < 10?9). Our data are suggestive of a novel mechanism by which monocytic AML cells evade cell-mediated immunity.

Aurelius, Johan; Thoren, Fredrik B.; Akhiani, Ali A.; Brune, Mats; Palmqvist, Lars; Hansson, Markus; Martner, Anna



Virus-specific HLA-restricted lysis of herpes simplex virus-infected human monocytes and macrophages mediated by cytotoxic T lymphocytes  

SciTech Connect

Freshly-isolated peripheral blood human monocytes and 5 day in vitro cultured macrophages were infected with herpes simplex virus type 1 (HSV-1), labeled with /sup 51/Cr, and used as target cells in a 12-14 hour cell-mediated cytotoxicity assay. Mononuclear leukocytes (MNL) from HSV-1 non-immune individuals, whether unstimulated or stimulated with HSV-1 antigen, did not mediate significant lysis of either target cell. HSV-immune MNL, both freshly-isolated and cultured for 5 days without antigen, demonstrated only low levels of natural killer (NK) cell-mediate lysis. MNL from HSV-immune individuals incubated for 5 days in vitro with HSV-1 antigen mediated significant virus-specific lysis of both target cells. Mean virus-specific lysis of autologous monocytes was 8.5(/+-/2.0)% compared to a three-fold greater virus-specific lysis of autologous macrophages. Greater than 70% of this lytic activity was mediated by Leu-11-negative, T3-positive cytotoxic T lymphocytes (CTL). Allogeneic target cells lacking a common HLA determinant were not significantly lysed while T8-positive CTL mediated infrequent lysis of target cells sharing a common HLA-A and/or HLA-B determinant. T4-positive lymphocytes were demonstrated to be the predominant cell mediating lysis of autologous target cells and allogeneic target cells sharing both HLA-A and/or HLA-B plus HLA-DR determinants with the CTL; the T4-positive cell was the sole CTL mediator of lysis of allogeneic target cells having a common HLA-DR determinant.

Torpey, D.J. III



Monocyte function in the acquired immune deficiency syndrome. Defective chemotaxis.  

PubMed Central

The ineffective immune response in patients with the acquired immune deficiency syndrome (AIDS) contributes to severe and widespread infections and unrestricted growth by certain tumors. To determine whether monocyte dysfunction contributes to this immunosuppressed condition, we investigated monocyte chemotaxis in patients with AIDS. Using three different chemotactic stimuli, N-formylmethionylleucylphenylalanine, lymphocyte-derived chemotactic factor, and C5a des Arg, we studied the chemotactic responses of monocytes from seven homosexual men with AIDS, three homosexuals with lymphadenopathy and an abnormal immunological profile, seven healthy homosexual men, and 23 heterosexual control individuals. Monocytes from each of the AIDS patients with Kaposi's sarcoma and/or opportunistic infection exhibited a marked reduction in chemotaxis to all stimuli compared with the healthy control subjects. The reduced chemotactic responses were observed over a wide range of concentrations for each stimulus. Monocytes from AIDS patients who had clinically apparent opportunistic infection(s) exhibited a greater reduction in monocyte migration to all three stimuli than monocytes from the AIDS patient with only Kaposi's sarcoma. Monocytes from each of three homosexuals with lymphadenopathy and an abnormal immunological profile exhibited decreased chemotactic responses that were intermediate between those of the AIDS patients and the healthy heterosexual control subjects. In contrast to these findings, monocytes from each of seven healthy homosexuals exhibited normal chemotactic responses to the same stimuli. In addition, monocytes from AIDS patients exhibited reduced chemotaxis to soluble products of Giardia lamblia, one of several protozoan parasites prevalent in AIDS patients. Thus the immune abnormality in AIDS, previously thought to involve only the T-, B-, and natural killer lymphocytes, extends to the monocyte-macrophage. Defective monocyte migratory function may contribute to the depressed inflammatory response to certain organisms and to the apparent unrestricted growth of certain neoplasms in patients with AIDS. Images

Smith, P D; Ohura, K; Masur, H; Lane, H C; Fauci, A S; Wahl, S M



Allergy or Tolerance: Reduced Inflammatory Cytokine Response and Concomitant IL-10 Production of Lymphocytes and Monocytes in Symptom-Free Titanium Dental Implant Patients  

PubMed Central

Hypersensitivity reactions to titanium (Ti) are very rare. Thus, we assessed the proinflammatory response and also potential tolerance favoring in vitro reactivity of human blood lymphocytes and monocytes (PBMC) to Ti in healthy individuals (14 without, 6 with complication-free dental Ti implants). The proliferation index (SI) in lymphocyte transformation test (LTT) and production of cytokines linked to innate immune response (IL-1?, IL-6, and TNF?) or immune regulation (IL-10) were assessed in response to TiO2 particles or Ti discs. In both groups, the Ti-LTT reactivity was not enhanced (e.g., SI < 3). The control antigen tetanus toxoid (TT) gave adequate reactivity (median SI individuals without/with implant: 20.6 ± 5.97/19.58 ± 2.99). Individuals without implant showed higher cytokine response to Ti materials than individuals with symptom-free implants; for example, TiO2 rutile particle induced increase of IL-1? 70.27-fold/8.49-fold versus control medium culture. PBMC of 5 of the 6 individuals with complication-free Ti implants showed an ex vivo ongoing production of IL-10 (mean 4.18 ± 2.98?pg/mL)-but none of the 14 controls showed such IL-10 production. Thus in vitro IL-1?-, IL-6-, and TNF-? production reflects “normal” unspecific immune response to Ti. This might be reduced by production of tolerogenic IL-10 in individuals with symptom-free Ti dental implants.

Thomas, Peter; Wollenberg, Andreas



Intraglomerular T cells and monocytes in nephritis: Study with monoclonal antibodies  

Microsoft Academic Search

Glomerular T cells and monocytes in nephritis: study with monoclonal antibodies. Intraglomerular T cells, monocytes, total leucocytes and other mononuclear subsets were sought in renal biopsies from patients with glomerulonephritis, using monoclonal antibodies and immunoperoxidase techniques. Twenty–four biopsies with no significant glomerular proliferation on optical microscopy, thirty–two with only endocapillary hypercellularity, and twenty–one with extra capillary crescentic glomerular disease were

Fernando E B Nolasco; John Stewart Cameron; Barrie Hartley; Adolfo Coelho; Gillian Hildreth; Rowena Reuben



Activated T lymphocytes suppress osteoclastogenesis by diverting early monocyte/macrophage progenitor lineage commitment towards dendritic cell differentiation through down-regulation of receptor activator of nuclear factor-kappaB and c-Fos  

PubMed Central

Activated T lymphocytes either stimulate or inhibit osteoclastogenesis from haematopoietic progenitors in different experimental models. To address this controversy, we used several modes of T lymphocyte activation in osteoclast differentiation ? mitogen-pulse, anti-CD3/CD28 stimulation and in vivo and in vitro alloactivation. Osteoclast-like cells were generated from non-adherent immature haematopoietic monocyte/macrophage progenitors in murine bone-marrow in the presence of receptor activator of nuclear factor (NF)-?B ligand (RANKL) and monocyte–macrophage colony-stimulating factor (M-CSF). All modes of in vivo and in vitro T lymphocyte activation and both CD4+ and CD8+ subpopulations produced similar inhibitory effects on osteoclastogenesis paralleled by enhanced dendritic cell (DC) differentiation. Osteoclast-inhibitory effect was associated with T lymphocyte activation and not proliferation, and could be replaced by their culture supernatants. The stage of osteoclast differentiation was crucial for the inhibitory action of activated T lymphocytes on osteoclastogenesis, because the suppressive effect was visible only on early osteoclast progenitors but not on committed osteoclasts. Inhibition was associated specifically with increased granulocyte–macrophage colony-stimulating factor (GM-CSF) expression by the mechanism of progenitor commitment toward lineages other than osteoclast because activated T lymphocytes down-regulated RANK, CD115, c-Fos and calcitonin receptor expression, and increased differentiation towards CD11c-positive DC. An activated T lymphocyte inhibitory role in osteoclastogenesis, confirmed in vitro and in vivo, mediated through GM-CSF release, may be used to counteract activated bone resorption mediated by T lymphocyte-derived cytokines in inflammatory and immune disorders. We also demonstrated the importance of alloactivation in osteoclast differentiation and the ability of cyclosporin A to abrogate T lymphocyte inhibition of osteoclastogenesis, thereby confirming the functional link between alloreaction and bone metabolism.

Grcevic, D; Lukic, I K; Kovacic, N; Ivcevic, S; Katavic, V; Marusic, A



Altered monocyte activation markers in Tourette's syndrome: a case-control study  

PubMed Central

Background Infections and immunological processes are likely to be involved in the pathogenesis of Tourette’s syndrome (TS). To determine possible common underlying immunological mechanisms, we focused on innate immunity and studied markers of inflammation, monocytes, and monocyte-derived cytokines. Methods In a cross-sectional study, we used current methods to determine the number of monocytes and levels of C-reactive protein (CRP) in 46 children, adolescents, and adult patients suffering from TS and in 43 healthy controls matched for age and sex. Tumor necrosis factor alpha (TNF-alpha), interleukin 6 (IL-6), soluble CD14 (sCD14), IL1-receptor antagonist (IL1-ra), and serum neopterin were detected by immunoassays. Results We found that CRP and neopterin levels and the number of monocytes were significantly higher in TS patients than in healthy controls. Serum concentrations of TNF-alpha, sIL1-ra, and sCD14 were significantly lower in TS patients. All measured values were within normal ranges and often close to detection limits. Conclusions The present results point to a monocyte dysregulation in TS. This possible dysbalance in innate immunity could predispose to infections or autoimmune reactions.



Induction of interleukin-1-beta release from human monocytes by cotton bract tannin.  


The human T lymphocyte proliferative response to cotton bract tannin was shown to be dependent upon the presence of monocytes. Since monocytes support the T cell mitogenic response by interleukin-1 (IL-1) production, it was anticipated that tannin has IL-1-inducing ability. To examine this possibility, human monocytes were cultured alone or with peripheral blood T lymphocytes, and stimulated with tannin. Control cultures included unstimulated cells, and cells challenged with other IL-1 inducers: concanavalin A (Con A) and lipopolysaccharide from Escherichia coli or Enterobacter agglomerans. IL-1 beta was measured in culture supernatants 24 h after initiation of the culture by the use of an ELISA or an RIA. The results showed that tannin stimulated monocytes to secrete IL-1 beta in a manner similar to Con A, i.e. substantially more cytokine was measured in the supernatants of monocyte-T-lymphocyte co-cultures than in the cultures of monocyte alone. Endotoxin from E. coli was less effective than the endotoxin from E. agglomerans in IL-1 induction. Contaminating endotoxin present in the tannin preparation accounted for the majority of IL-1 beta released from monocytes alone stimulated with tannin, but only 20% of the IL-1 beta released from tannin-stimulated monocyte-T-lymphocyte co-cultures. These results show that tannin itself has IL-1-inducing ability. The dose-response studies show that the extent of IL-1 beta release is dependent on tannin dose and that increased levels of monocyte-produced IL-1 beta precede the increase in T lymphocyte proliferation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2210874

Vuk-Pavlovi?, Z; Rohrbach, M S



Impact of Human Granulocyte and Monocyte Isolation Procedures on Functional Studies  

PubMed Central

One of the first lines of defense against infection is the activation of the innate immune system. It is becoming clear that autoimmune diseases, such as rheumatoid arthritis and Crohn's disease, may be caused by disturbed innate immunity, and relating granulocyte and monocyte functions to the patient genotype has become an important part of contemporary research. Although it is essential to move this field forward, a systematic study comparing the efficacy and suitability for functional studies of the various available protocols for the isolation of these immune cells has not been performed. Here, we compare human granulocyte functionality under three enrichment protocols: (i) Ficoll density gradient centrifugation, (ii) anti-CD15 antibody-conjugated microbeads (positive selection), and (iii) Polymorphoprep. Primary monocytes were isolated in parallel using (i) anti-CD14 magnetic microbeads, (ii) non-monocyte depletion by antibody-conjugated magnetic microbeads (negative selection), (iii) RosetteSep antibody cocktail, and (iv) the classical adherence protocol. The best results in terms of purity and cell functionality were obtained with positive selection by magnetic microbeads for both human granulocytes and monocytes. Whereas phagocytosis of Escherichia coli bacteria was identical in all isolation procedures tested, the granulocyte respiratory burst was higher in positively selected cells. In addition, different granulocyte enrichment procedures affect cell surface receptor expression to different extents. In toto, we propose that positive selection of granulocytes and monocytes be adopted as the procedure of choice for studies of human granulocyte and monocyte functions but caution investigators to be aware of possible alterations in cell phenotypes with different isolation procedures.

Zhou, Lu; Somasundaram, Rajesh; Nederhof, Rosa F.; Dijkstra, Gerard; Faber, Klaas Nico; Peppelenbosch, Maikel P.



Impact of human granulocyte and monocyte isolation procedures on functional studies.  


One of the first lines of defense against infection is the activation of the innate immune system. It is becoming clear that autoimmune diseases, such as rheumatoid arthritis and Crohn's disease, may be caused by disturbed innate immunity, and relating granulocyte and monocyte functions to the patient genotype has become an important part of contemporary research. Although it is essential to move this field forward, a systematic study comparing the efficacy and suitability for functional studies of the various available protocols for the isolation of these immune cells has not been performed. Here, we compare human granulocyte functionality under three enrichment protocols: (i) Ficoll density gradient centrifugation, (ii) anti-CD15 antibody-conjugated microbeads (positive selection), and (iii) Polymorphoprep. Primary monocytes were isolated in parallel using (i) anti-CD14 magnetic microbeads, (ii) non-monocyte depletion by antibody-conjugated magnetic microbeads (negative selection), (iii) RosetteSep antibody cocktail, and (iv) the classical adherence protocol. The best results in terms of purity and cell functionality were obtained with positive selection by magnetic microbeads for both human granulocytes and monocytes. Whereas phagocytosis of Escherichia coli bacteria was identical in all isolation procedures tested, the granulocyte respiratory burst was higher in positively selected cells. In addition, different granulocyte enrichment procedures affect cell surface receptor expression to different extents. In toto, we propose that positive selection of granulocytes and monocytes be adopted as the procedure of choice for studies of human granulocyte and monocyte functions but caution investigators to be aware of possible alterations in cell phenotypes with different isolation procedures. PMID:22552601

Zhou, Lu; Somasundaram, Rajesh; Nederhof, Rosa F; Dijkstra, Gerard; Faber, Klaas Nico; Peppelenbosch, Maikel P; Fuhler, Gwenny M



Cannabidiol induced a contrasting pro-apoptotic effect between freshly isolated and precultured human monocytes  

SciTech Connect

It has been documented that cannabidiol (CBD) induced apoptosis in a variety of transformed cells, including lymphocytic and monocytic leukemias. In contrast, a differential sensitivity between normal lymphocytes and monocytes to CBD-mediated apoptosis has been reported. The present study investigated the pro-apoptotic effect of CBD on human peripheral monocytes that were either freshly isolated or precultured for 72 h. CBD markedly enhanced apoptosis of freshly isolated monocytes in a time- and concentration-dependent manner, whereas precultured monocytes were insensitive. By comparison, both cells were sensitive to doxorubicin-induced apoptosis. CBD significantly diminished the cellular thiols and glutathione in freshly isolated monocytes. The apoptosis induced by CBD was abrogated in the presence of N-acetyl-{sub L}-cysteine, a precursor of glutathione. In addition, precultured monocytes contained a significantly greater level of glutathione and heme oxygenase-1 (HO-1) compared to the freshly isolated cells. The HO-1 competitive inhibitor zinc protoporphyrin partially but significantly restored the sensitivity of precultured monocytes to CBD-mediated apoptosis. Collectively, our results demonstrated a contrasting pro-apoptotic effect of CBD between precultured and freshly isolated monocytes, which was closely associated with the cellular level of glutathione and the antioxidative capability of the cells.

Wu, Hsin-Ying; Chang, An-Chi; Wang, Chia-Chi; Kuo, Fu-Hua; Lee, Chi-Ya [Department and Graduate Institute of Veterinary Medicine, School of Veterinary Medicine, National Taiwan University, Taipei, Taiwan (China); Liu, Der-Zen [Graduate Institute of Biomedical Materials and Engineering, Taipei Medical University, Taipei, Taiwan (China); Jan, Tong-Rong, E-mail: [Department and Graduate Institute of Veterinary Medicine, School of Veterinary Medicine, National Taiwan University, Taipei, Taiwan (China)



RANTES Expression and Contribution to Monocyte Chemotaxis in Arthritis  

Microsoft Academic Search

Rheumatoid arthritis (RA) is characterized by recruitment of leukocytes from the vasculature into inflamed synovial tissue (ST) and synovial fluid (SF), which depends, in part, upon the continued maintenance of chemotactic stimuli. RANTES is a potent chemoattractant for leukocytes including monocytes and CD45RO+memory T lymphocytes. The aim of this study was to determine the production, the source, and the function

Michael V. Volin; Manisha R. Shah; Michihide Tokuhira; G. Kenneth Haines; James M. Woods; Alisa E. Koch



Studies on cytolytic mechanisms of activated macrophages and monocytes  

SciTech Connect

Pulmonary alveolar macrophages (PAM) and peripheral blood monocytes (PBMo) of the miniature swine can be converted to cytolytically active cells by treating with phorbol myristic acetate (PMA) or combined stimulation with recombinant human interferon - ..gamma.. (rHuIFN-..gamma..) and lipopolysaccharide (LPS). These activated PAM and PBMo become cytolytic to various targets by producing neutral proteases. H/sub 2/O/sub 2/ and cytotoxic factors. While the PAM stimulated by PMA was most active in generating H/sub 2/O/sub 2/ as well as neutral proteases, the PBMo stimulated with PMA was able to produce only H/sub 2/O/sub 2/. The mechanism of killing by PAM seemed to vary with type of target cells: H/sub 2/O/sub 2/ was effective in killing of PRBC, SRBC, and K562, but proteases were responsible for the lysis of U937 and WEHI-164. In contrast to PMA stimulation, combined stimulation with rHuIFN-..gamma.. and LPS was very effective in generation of cytotoxic factors from PAM and PBMo. These cytotoxic factors were directly toxic to various target cells including WEHI-164, U937, K562, and CRBC, but not to autologous target such as PRBC.

Chung, T.; Kim, Y.B.



Influence of therapy with chimeric monoclonal tumour necrosis factor-a antibodies on intracellular cytokine profiles of T lymphocytes and monocytes in rheumatoid arthritis patients  

Microsoft Academic Search

Introduction. It has been shown that T lymphocytes and monocytesumacrophages, producing pro-inflammatory cytokines, play a pivotal role in the pathophysio- logy of rheumatoid arthritis (RA). In recent placebo-controlled double-blind randomized studies, chimeric (humanumouse) tumour necrosis factor-a (TNFa) antibodies (cA2) proved to be very effective in improving clinical disease activity and reducing inflammatory parameters in RA. Objective. To investigate whether anti-TNFa

A. J. Schuerwegh; J. F. Van Offel; W. J. Stevens; C. H. Bridts; L. S. De Clerck



Decreased monocyte antibody-dependent cell-mediated toxicity in stage I–II malignant melanoma  

Microsoft Academic Search

Lymphocyte and monocyte antibody-dependent cell-mediated cytotoxicity (ADCC) against human red blood cells was examined in 28 stage-I-II malignant melanoma patients. Eighteen were studied at various time intervals after receiving SC Corynebacterium parvum (C. parvum); 10 were untreated. Fifteen normal age-matched controls were also studied. Monocyte ADCC was significantly decreased in untreated patients compared with controls (P<0.005) and was significantly increased

James L. Murray; Elisa T. Lee



Lysis of uninfected HIV1 gp120-coated peripheral blood-derived T lymphocytes by monocyte-mediated antibody-dependent cellular cytotoxicity  

Microsoft Academic Search

Previous reports from our laboratory have demonstrated that peripheral blood monocytes (PBM) from HIV-1 infected individuals are de novo activated and are cytotoxic in vitro. Significant monocyte-antibody-dependent cellular cytotoxicity (ADCC) was obtained against HIV-1 inactivated CD4+ CEM target cells coated with HIV-1 in the presence of autologous seropositive serum. Based on these findings, we hypothesized that in HIV-seropositive individuals the

Didier Hober; Anahid Jewett; Benjamin Bonavida



Calorimetric studies of the state of water in deeply frozen human monocytes.  


Intra- and extracellular phase transitions in human peripheral blood monocyte suspensions with and without the cryoprotectant 1 M dimethylsulfoxide were measured using differential scanning calorimetry. Using an fluorescence diacetate/ethidium bromide assay for membrane integrity and a phagocytosis assay for cell function, it was found that mortality was correlated with several phase transitions under a variety of cooling and warming regimens. As a result of these studies we concluded that: intracellular freezing is lethal, but avoidance of freezing during fast cooling is not sufficient to provide complete protection; a subtle freezing injury in the cryoprotected monocytes can be correlated with a measurable increase in devitrification on warming; and the cell contents form more stable glasses than the Hanks' balanced salt solution with fetal calf serum used as the extracellular medium. PMID:3978207

Takahashi, T; Hirsh, A



Calorimetric studies of the state of water in deeply frozen human monocytes.  

PubMed Central

Intra- and extracellular phase transitions in human peripheral blood monocyte suspensions with and without the cryoprotectant 1 M dimethylsulfoxide were measured using differential scanning calorimetry. Using an fluorescence diacetate/ethidium bromide assay for membrane integrity and a phagocytosis assay for cell function, it was found that mortality was correlated with several phase transitions under a variety of cooling and warming regimens. As a result of these studies we concluded that: intracellular freezing is lethal, but avoidance of freezing during fast cooling is not sufficient to provide complete protection; a subtle freezing injury in the cryoprotected monocytes can be correlated with a measurable increase in devitrification on warming; and the cell contents form more stable glasses than the Hanks' balanced salt solution with fetal calf serum used as the extracellular medium.

Takahashi, T; Hirsh, A



Magnetic resonance spectroscopy reveals that activated monocytes contribute to neuronal injury in SIV neuroAIDS  

PubMed Central

Difficulties in understanding the mechanisms of HIV neuropathogenesis include the inability to study dynamic processes of infection, cumulative effects of the virus, and contributing host immune responses. We used 1H magnetic resonance spectroscopy and studied monocyte activation and progression of CNS neuronal injury in a CD8 lymphocyte depletion model of neuroAIDS in SIV-infected rhesus macaque monkeys. We found early, consistent neuronal injury coincident with viremia and SIV infection/activation of monocyte subsets and sought to define the role of plasma virus and monocytes in contributing to CNS disease. Antiretroviral therapy with essentially non–CNS-penetrating agents resulted in slightly decreased levels of plasma virus, a significant reduction in the number of activated and infected monocytes, and rapid, near-complete reversal of neuronal injury. Robust macrophage accumulation and productive virus replication were found in brains of infected and CD8 lymphocyte–depleted animals, but no detectable virus and few scattered infiltrating macrophages were observed in CD8 lymphocyte–depleted animals compared with animals not receiving antiretroviruses that were sacrificed at the same time after infection. These results underscore the role of activated monocytes and monocyte infection outside of the brain in driving CNS disease.

Williams, Kenneth; Westmoreland, Susan; Greco, Jane; Ratai, Eva; Lentz, Margaret; Kim, Woong-Ki; Fuller, Robert A.; Kim, John P.; Autissier, Patrick; Sehgal, Prahbat K.; Schinazi, Raymond F.; Bischofberger, Norbert; Piatak, Michael; Lifson, Jeffrey D.; Masliah, Eliezer; Gonzalez, R. Gilberto



An in vitro study of the effect of size and timing of administration of titanium dioxide particles on antigen presenting activity of alveolar macrophages and peripheral blood monocytes.  


Previous studies have shown that inhaled particles exacerbate asthma and allergic rhinitis. Several factors related to the particle may play a role in immune-stimulating activity; however, the underlying mechanisms remain unclear. We carried out in vitro studies to investigate the effects of TiO(2) particle exposure on antigen presenting activity and expression of the associated cell-surface molecules (Ia, B7.1, B7.2) in rat derived monocytes and alveolar macrophages, in terms of two aspects of the particles: (1) size (59 nm (ST) and 350 nm (LT) particles), and (2) the timing of particle exposure (before antigen exposure or co-administered). Results indicated that particle exposure prior to antigen exposure led to decreased antigen presenting activity in both types of cell. This decrease was greater with ST particles. In monocytes, the expression of cell surface molecules decreased similarly with both particles. Conversely, alveolar macrophages showed greater expression of Ia with ST than with LT exposures. Ia expression was confirmed to be functionally active by a mixed lymphocyte reaction. It is possible that particle exposure might result in poor antigen processing, thereby leading to decreased antigen presenting activity. Co-exposure of particles and antigen induced an increase in antigen presenting activity with both types of particle; however, ST exposure induced greater antigen presenting activity. The expression of Ia also increased similarly with both particle sizes. This suggests that, in a co-exposure situation, antigen may be processed without intensive retardation by particles, and factors other than Ia may affect antigen presenting activity. In conclusion, both size and timing of exposure to TiO(2) particles affect antigen presenting activity of monocytes and alveolar macrophages. PMID:19653805

Munidasa, Dulee Tamirei; Koike, Eiko; Kobayashi, Takahiro



Microenvironments of T and B lymphocytes : a light- and electromicroscopic study  

Microsoft Academic Search

Peripheral blood cells- erythrocytes, granulocytes, monocytes, thrombocytes\\u000aand lymphocytes-are the end products of a differentiation process which\\u000aoccurs in the bone marrow and, in rodents, also in the spleen. Normal\\u000ahaemopoietic tissue is a cell renewal system with an accurate balance between\\u000acell production originating from pluripotent haemopoietic stem cells and\\u000acontinuous cell loss. The important function of haemopoietic stem

Ewijk van W



Phenotype and function of monocyte derived dendritic cells in chronic hepatitis B virus infection  

Microsoft Academic Search

The antiviral T cell failure of patients with chronic hepatitis B virus (HBV) infection was suggested to be caused by a T cell stimulation defect of dendritic cells (DC). To address this hypothesis, monocyte derived DC(MDDC) of patients withchronic or resolvedacute HBVinfection and healthy controls were studied phenotypically by FACS analyses and functionally by mixed lymphocyte reaction, ELISA, ELISpot and

Soheila Tavakoli; Wibke Schwerin; Andreas Rohwer; Sina Hoffmann; Sandra Weyer; Robert Weth; Helga Meisel; Helmut Diepolder; Michael Geissler; Peter R. Galle; Hanns F. Lohr; Wulf O. Bocher



Studies of Lymphocyte Growth and Differentiation. Progress Report, April 15, 1974--April 1, 1975.  

National Technical Information Service (NTIS)

Progress is reported on studies of growth in cultured lymphocytes in which it was demonstrated that ribonuclear protein maturation in lymphocyte nucleoli depends on the interaction between nascent RNA and proteins and that phytohemagglutinin (PHA) can dir...

A. D. Rubin



Studies of lymphocyte growth and differentiation. Progress report, September 1, 1975July 31, 1976  

Microsoft Academic Search

Studies were continued on ribonuclear protein synthesis and the assembly of ribosomes in resting and stimulated lymphocytes. We demonstrated the interdependency of protein synthesis and RNA synthesis in the formation and processing of nascent ribonuclear protein particles. We further explored lymphocyte nuclei in a cell-free system. By isolating lymphocyte chromatin we showed a direct effect of PHA on the ability




An innate response to allogeneic nonself mediated by monocytes.  


The mammalian innate immune system has evolved diverse strategies to distinguish self from microbial nonself. How the innate immune system distinguishes self-tissues from those of other members of the same species (allogeneic nonself) is less clear. To address this question, we studied the cutaneous hypersensitivity response of lymphocyte-deficient RAG(-/-) mice to spleen cells transplanted from either allogeneic or syngeneic RAG(-/-) donors. We found that RAG(-/-) mice mount a specific response to allogeneic cells characterized by swelling and infiltration of the skin with host monocytes/macrophages and neutrophils. The response required prior priming with allogeneic splenocytes or skin grafts and exhibited features of memory as it could be elicited at least 4 wk after immunization. Neither depletion of host NK cells nor rechallenging immunized mice with F(1) hybrid splenocytes inhibited the response, indicating that the response is not mediated by NK cells. Depletion of host monocytes/macrophages or neutrophils at the time of rechallenge significantly diminished the response and, importantly, the adoptive transfer of monocytes from alloimmunized RAG(-/-) mice conferred alloimmunity to naive RAG(-/-) hosts. Unlike NK- and T cell-dependent alloresponses, monocyte-mediated alloimmunity could be elicited only when donor and responder mice differed at non-MHC loci. These observations indicate that monocytes mount a response to allogeneic nonself, a function not previously attributed to them, and suggest the existence of mammalian innate allorecognition strategies distinct from detection of missing self-MHC molecules by NK cells. PMID:19923456

Zecher, Daniel; van Rooijen, Nico; Rothstein, David M; Shlomchik, Warren D; Lakkis, Fadi G




Microsoft Academic Search

The scqucntial transformation of chickcn monocytcs into macrophages, cpithelioid cells, and multinucleatcd giant cells in vitro was studied by electron microscopy after fixation and cmbcdment in situ. The following changes occur. In the nucleus, margination of chro- matin, cvidcnt in monocytes, decreases in later forms. Nucleoli become more complcx and nuclear pores increase in number. In cytoplasm, a progressive increase




Monocyte-derived dendritic cells loaded with a mixture of apoptotic\\/necrotic melanoma cells efficiently cross-present gp100 and MART1 antigens to specific CD8+ T lymphocytes  

Microsoft Academic Search

BACKGROUND: In the present study, we demonstrate, in rigorous fashion, that human monocyte-derived immature dendritic cells (DCs) can efficiently cross-present tumor-associated antigens when co-cultured with a mixture of human melanoma cells rendered apoptotic\\/necrotic by ? irradiation (Apo-Nec cells). METHODS: We evaluated the phagocytosis of Apo-Nec cells by FACS after PKH26 and PKH67 staining of DCs and Apo-Nec cells at different

Erika M von Euw; María M Barrio; David Furman; Michele Bianchini; Estrella M Levy; Cassian Yee; Yongqing Li; Rosa Wainstok; José Mordoh



Prion protein induced signaling cascades in monocytes  

SciTech Connect

Prion proteins play a central role in transmission and pathogenesis of transmissible spongiform encephalopathies. The cellular prion protein (PrP{sup C}), whose physiological function remains elusive, is anchored to the surface of a variety of cell types including neurons and cells of the lymphoreticular system. In this study, we investigated the response of a mouse monocyte/macrophage cell line to exposure with PrP{sup C} fusion proteins synthesized with a human Fc-tag. PrP{sup C} fusion proteins showed an attachment to the surface of monocyte/macrophages in nanomolar concentrations. This was accompanied by an increase of cellular tyrosine phosphorylation as a result of activated signaling pathways. Detailed investigations exhibited activation of downstream pathways through a stimulation with PrP fusion proteins, which include phosphorylation of ERK{sub 1,2} and Akt kinase. Macrophages opsonize and present antigenic structures, contact lymphocytes, and deliver cytokines. The findings reported here may become the basis of understanding the molecular function of PrP{sup C} in monocytes and macrophages.

Krebs, Bjarne [Center for Neuropathology and Prion Research, Ludwig-Maximilians-University Munich, Muenchen (Germany); Dorner-Ciossek, Cornelia [CNS Research III, Boehringer Ingelheim Pharma GmbH and Co KG, Biberach/Riss (Germany); Schmalzbauer, Ruediger [Center for Neuropathology and Prion Research, Ludwig-Maximilians-University Munich, Munich (Germany); Vassallo, Neville [Department of Physiology and Biochemistry, University of Malta, Msida (Malta); Herms, Jochen [Center for Neuropathology and Prion Research, Ludwig-Maximilians-University Munich, Munich (Germany); Kretzschmar, Hans A. [Center for Neuropathology and Prion Research, Ludwig-Maximilians-University Munich, Munich (Germany)]. E-mail:



Study of monocyte membrane proteome perturbation during lipopolysaccharide-induced tolerance using iTRAQ-based quantitative proteomic approach.  


Human monocytes' exposure to low-level lipopolysaccharide (LPS) induces temporary monocytic insensitivity to subsequent LPS challenge. The underlying mechanism of this phenomenon could have important clinical utilities in preventing and/or treating severe infections. In this study, we used an iTRAQ-based quantitative proteomic approach to comprehensively characterize the membrane proteomes of monocytes before and after LPS exposure. We identified a total of 1651 proteins, of which 53.6% were membrane proteins. Ninety-four percent of the proteins were quantified and 255 proteins were shown to be tightly regulated by LPS. Subcellular location analysis revealed organelle-specific response to LPS exposure: more than 90% of identified mitochondrial membrane proteins were significant downregulated, whereas the majority of proteins from other organelles such as ER, Golgi and ribosome were upregulated. Moreover, we found that the expression of most receptors potentially involved in LPS signal pathway (CD14, toll-like receptor 4, CD11/CD18 complex) were substantially decreased, while the expression of molecules involved in LPS neutralization were enhanced after LPS challenge. Together, these findings could be of significance in understanding the mechanism of LPS tolerance and provide values for designing new approaches for regulating monocytic responses in sepsis patients. PMID:20486119

Zhang, Huoming; Zhao, Changqing; Li, Xin; Zhu, Yi; Gan, Chee Sian; Wang, Yong; Ravasi, Timothy; Qian, Pei-Yuan; Wong, Siew Cheng; Sze, Siu Kwan



Effect of salmonella-infected human monocytes on natural killer cell cytotoxicity. In vitro studies  

Microsoft Academic Search

Various chemicals, including some bacteria-derived components, modulate natural killer cell (NKC) activity. We have analyzed the effect of wild-type Ty2 and of mutant strain TYT1231 Salmonella typhi-infected monocytes (U937 cells and human autologous monocytes) on NKC cytotoxicity of peripheral blood mononuclear cell (PBMC) and highly purified NKC (HPNKC; CD16+\\/56+>95%; the rest corresponding to CD3+ T-cells).PBMC's co-culture with either S. typhi

Luz Blanco; Javier Puente; Carolina Carrasco; Dante Miranda; Marion E Wolf; Aron D Mosnaim



Collagen XIII Induced in Vascular Endothelium Mediates ?1?1 Integrin-Dependent Transmigration of Monocytes in Renal Fibrosis  

PubMed Central

Alport syndrome is a common hereditary basement membrane disorder caused by mutations in the collagen IV ?3, ?4, or ?5 genes that results in progressive glomerular and interstitial renal disease. Interstitial monocytes that accumulate in the renal cortex from Alport mice are immunopositive for integrin ?1?1, while only a small fraction of circulating monocytes are immunopositive for this integrin. We surmised that such a disparity might be due to the selective recruitment of ?1?1-positive monocytes. In this study, we report the identification of collagen XIII as a ligand that facilitates this selective recruitment of ?1?1 integrin-positive monocytes. Collagen XIII is absent in the vascular endothelium from normal renal cortex and abundant in Alport renal cortex. Neutralizing antibodies against the binding site in collagen XIII for ?1?1 integrin selectively block VLA1-positive monocyte migration in transwell assays. Injection of these antibodies into Alport mice slows monocyte recruitment and protects against renal fibrosis. Thus, the induction of collagen XIII in endothelial cells of Alport kidneys mediates the selective recruitment of ?1?1 integrin-positive monocytes and may potentially serve as a therapeutic target for inflammatory diseases in which lymphocyte/monocyte recruitment involves the interaction with ?1?1 integrin.

Dennis, Jameel; Meehan, Daniel T.; Delimont, Duane; Zallocchi, Marisa; Perry, Greg A.; O'Brien, Stacie; Tu, Hongmin; Pihlajaniemi, Taina; Cosgrove, Dominic



CD4-Specific Transgenic Expression of Human Cyclin T1 Markedly Increases Human Immunodeficiency Virus Type 1 (HIV-1) Production by CD4+ T Lymphocytes and Myeloid Cells in Mice Transgenic for a Provirus Encoding a Monocyte-Tropic HIV-1 Isolate  

PubMed Central

Human immunodeficiency virus type 1 (HIV-1)-encoded Tat provides transcriptional activation critical for efficient HIV-1 replication by interacting with cyclin T1 and recruiting P-TEFb to efficiently elongate the nascent HIV transcript. Tat-mediated transcriptional activation in mice is precluded by species-specific structural differences that prevent Tat interaction with mouse cyclin T1 and severely compromise HIV-1 replication in mouse cells. We investigated whether transgenic mice expressing human cyclin T1 under the control of a murine CD4 promoter/enhancer cassette that directs gene expression to CD4+ T lymphocytes and monocytes/macrophages (hu-cycT1 mice) would display Tat responsiveness in their CD4-expressing mouse cells and selectively increase HIV-1 production in this cellular population, which is infected primarily in HIV-1-positive individuals. To this end, we crossed hu-cycT1 mice with JR-CSF transgenic mice carrying the full-length HIV-1JR-CSF provirus under the control of the endogenous HIV-1 long terminal repeat and demonstrated that human cyclin T1 expression is sufficient to support Tat-mediated transactivation in primary mouse CD4 T lymphocytes and monocytes/macrophages and increases in vitro and in vivo HIV-1 production by these stimulated cells. Increased HIV-1 production by CD4+ T lymphocytes was paralleled with their specific depletion in the peripheral blood of the JR-CSF/hu-cycT1 mice, which increased over time. In addition, increased HIV-1 transgene expression due to human cyclin T1 expression was associated with increased lipopolysaccharide-stimulated monocyte chemoattractant protein 1 production by JR-CSF mouse monocytes/macrophages in vitro. Therefore, the JR-CSF/hu-cycT1 mice should provide an improved mouse system for investigating the pathogenesis of various aspects of HIV-1-mediated disease and the efficacies of therapeutic interventions.

Sun, Jinglin; Soos, Timothy; KewalRamani, Vineet N.; Osiecki, Kristin; Zheng, Jian Hua; Falkin, Laurie; Santambrogio, Laura; Littman, Dan R.; Goldstein, Harris



Alternate-day prednisone therapy and human lymphocyte subpopulations.  

PubMed Central

The mechanisms and kinetics of the immunosuppressive effects of alternate-day prednisone were investigated in a group of patients with a variety of inflammatory diseases receiving a range of alternate-day prednisone doses from 5 to 120 mg. Total circulating lymphocyte and monocyte counts, as well as proportions of lymphocyte subpopulations defined both by surface markers and by in vitro functional capacities, were studied. At 8 a. m. of the day on prednisone, just before drug administration, lymphocyte and monocyte counts, proportions of lymphocyte subpopulations, as well as in vitro lymphocyte blastogenic responses to various mitogenic and antigenic stimuli were normal. 4 h after the administration of prednisone, there was a profound lymphocytopenia and monocytopenia, with a differential depletion of thymus-derived lymphocytes as well as various functionally defined lymphocyte subpopulations. Lymphocyte kinetic studies using a radioactive chromium-labeled autologous lymphocytes showed that the lymphocytopenia was due predominantly to a transient depletion of the recirculating portion of the intravascular lymphocytepool. All these parameters returned to normal by 8 a.m. of the following day (off prednisone) and remained normal throughout the day. This very transient lymphocytopenia and monocytopenia after prednisone, with normal cell numbers, proportions, and functions throughout the remainder of the 2-day cycle, was associated with suppression of disease activity, yet did not affect cutaneous delayed hypersensitivity in these patients nor increase the likelihood of infectious complications. This drug-associated cyclic and transient monocytopenia and selective lymphocytopenia is best explained by a redistribution of recirculating lymphocytes to other body compartments, particularly the bone marrow.

Fauci, A S; Dale, D C



Surface markers on human lymphocytes: studies of normal subjects and of patients with primary immunodeficiencies  

PubMed Central

Peripheral blood lymphocytes of twenty normal controls and twelve patients with primary immunodeficiencies were examined for surface membrane Ig and receptors for C3 complement (B cell markers) and for spontaneous rosette formation with sheep erythrocytes (T cell markers). In patients with defects in T cell function no lymphocytes forming spontaneous rosettes were seen. In patients with B cell deficiency they were normal or increased. Lymphocytes with membrane immunoglobulins were normal in patients with T cell defect and absent in patients with severe agammaglobulinaemia. Lymphocytes with receptors for C3 complement were increased in patients with T defect and normal in patients with most other forms of immunodeficiency studied.

Aiuti, F.; Lacava, V.; Garofalo, J. A.; D'Amelio, R.; D'Asero, C.



Tissue Culture Studies of Human Lymphocyte in Infectious Mononucleosis.  

National Technical Information Service (NTIS)

The site of production for the heterophile antibody in patients with infectious mononucleosis is unknown. Atypical lymphocytes from patients with infectious mononucleosis were grown in tissue culture in an attempt to demonstrate heterophile antibody produ...

R. A. Gams C. A. Coltman



'Long-distance' lymphocyte study. The effects of transporting blood in lymphocyte blastogenic responses.  

PubMed Central

A comparison of blastogenic responsiveness to antigens and mitogen by human lymphocytes was made between cells which had been processed for culture immediately following blood collection and cells obtained from blood collected 9-11 hr previously and transported via commercial airline from the patients' homes to our laboratory. There were no significant differences in the responses of transported and non-transported cells if the blood was maintained at ambient temperature during the period of shipment. Chilling the blood during transport, however, resulted both in decreased stimulation of the cells and increased 'background' activity in unstimulated cultures. These findings indicate the feasibility of carrying out both limited immunological evaluations and extended periods of follow-up for patients located at considerable distances from a research laboratory.

Ottesen, E A; Poindexter, R W; Hiatt, R A



Antiphospholipid antibody effects on monocytes  

Microsoft Academic Search

Although the presence of autoantibodies is known to increase the risk of thrombosis in the antiphospholipid syndrome, the\\u000a mechanism by which these antibodies exert their effects is poorly understood. Several studies suggest that autoantibody-mediated\\u000a dysregulation of monocytes is one pathobiologic mechanism of this disease. Recent studies have focused on extra-and intracellular\\u000a interactions involved in monocyte activation and expression of procoagulant

Alisa S. Wolberg



Plasmacytoid T cells. Immunohistochemical evidence for their monocyte/macrophage origin.  

PubMed Central

To elucidate the lineage of plasmacytoid T cells, their immunophenotype was studied in reactive lymph nodes with a broad panel of monoclonal antibodies. Plasmacytoid T cells expressed several myelomonocytic markers, and almost all markers highly selective for macrophages. They lacked granulocyte-associated and B or T lymphocyte-associated antigens. These results provide strong evidence that plasmacytoid T cells are of monocyte lineage. Images Figure 1 Figure 2

Facchetti, F.; de Wolf-Peeters, C.; Mason, D. Y.; Pulford, K.; van den Oord, J. J.; Desmet, V. J.



The interleukin-6 promoter polymorphism is associated with elevated leukocyte, lymphocyte, and monocyte counts and reduced physical fitness in young healthy smokers  

Microsoft Academic Search

Smoking and interleukin-6 are important factors in driving inflammation. This study assessed the relationship between smoking, interleukin-6 genotype, physical fitness, and peripheral blood count in healthy young men. For this interleukin-6 promoter polymorphism -174 genotype-phenotype association study 1,929 healthy German male aviators recruited at the central German Air Force Institute of Aviation Medicine were stratified by smoking habits. Cardiovascular fitness

J. R. Ortlepp; J. Metrikat; K. Vesper; V. Mevissen; F. Schmitz; M. Albrecht; P. Maya-Pelzer; P. Hanrath; C. Weber; K. Zerres; R. Hoffmann



Immunologic Effector Mechanisms of a Standardized Mistletoe Extract on the Function of Human Monocytes and Lymphocytes in vitro , ex vivo , and in vivo  

Microsoft Academic Search

Even though mistletoe extracts have been in clinical use for centuries their exact mode of action is still unknown. Currently, the application scheme for registered preparations is a dose-escalating scheme to thus reduce side effects. In this study, healthy controls and patients were evaluated for their immunologic response to treatment with a standardized mistletoe extract (Iscador). It shows a strong

Lucie Heinzerling; Volker Von Baehr; Christa Liebenthal; RÜdiger Von Baehr; Hans-Dieter Volk




PubMed Central

The proliferative interaction of cultured rat lymphocytes of immunogenetically disparate origin—the mixed lymphocyte interaction—was employed as an experimental model to examine the initial stages of the immune response mechanism. Using mixed cultures of cells derived from parental strain and F1 hybrid rats, in which only the parental lymphocytes respond, the following observations were made on the magnitude and kinetics of the reaction. After initiation of the cultures, there was a latent period of approximately 40 hours during which time no mitotic activity was detected. This inactive phase was followed by a period of proliferation in which previously nondividing cells entered the mitotic cycle for the first time. Activity in the cultures, as detected by incorporation of radioactive thymidine and measured by radioautography or scintillation spectrometry, increased exponentially with a doubling time (T2) of 9–10 hr. In this exponential proliferative phase, lasting approximately 100 hr, the dividing cells underwent a series of rapid sequential divisions with a generation time (Tc) of 8 hr, and few, if any, dropped out of the mitotic cycle. In addition to the cells which first entered mitosis at the beginning of the proliferative phase and then proceeded through multiple divisions, significant numbers of new, previously nondividing cells continued to enter the mitotic cycle during the entire exponential growth phase. The total number of these newly responsive, first division cells throughout the total culture period amounted to 1–3% of the original parental cell inoculum. This is a surprisingly large proportion of peripheral blood lymphocytes with demonstrable reactivity to a particular antigen system, if it is assumed that these first division cells in vitro are functionally related to the hypothetical antigen-sensitive cells which proliferate and differentiate into immunological effector cells. At present there is no entirely satisfactory explanation for this large number of reactive cells in the mixed lymphocyte interaction.

Wilson, Darcy B.; Blyth, Janet L.; Nowell, Peter C.



UVB radiation and human monocyte accessory function: Differential effects on pre-mitotic events in T-cell activation  

SciTech Connect

Purified T lymphocytes fail to proliferate in response to antigenic and mitogenic stimuli when cultured in the presence of accessory cells that have been exposed in vitro to sublethal doses of UVB radiation. Because proliferation represents a final stage in the T-cell activation process, the present study was conducted to determine whether T cells were able to progress through any of the pre-mitotic stages when UVB-irradiated monocytes were used as model accessory cells. In these experiments, monoclonal anti-CD3 antibodies were employed as the mitogenic stimulus. Culture of T cells with UVB-irradiated monocytes did allow the T cells to undergo an increase in intracellular free calcium, which is one of the first steps in the activation sequence. The T cells expressed interleukin-2 receptors, although at a reduced level. However, T cells failed to produce interleukin-2 above background levels when they were placed in culture with monocytes exposed to UVB doses as low as 50 J/m2. Incubation of T cells with UVB-irradiated monocytes did not affect the subsequent capacity of T cells to proliferate, since they developed a normal proliferative response in secondary culture when restimulated with anti-CD3 antibodies and unirradiated monocytes. These studies indicate that T lymphocytes become partially activated when cultured with UVB-irradiated monocytes and mitogenic anti-CD3 monoclonal antibodies. In addition, they suggest that interleukin-2 production is the T-cell activation step most sensitive to inhibition when UVB-irradiated monocytes are employed as accessory cells.

Krutmann, J.K.; Kammer, G.M.; Toossi, Z.; Waller, R.L.; Ellner, J.J.; Elmets, C.A. (Case Western Reserve Univ., Cleveland, OH (USA))



Fifty cases of human immunodeficiency virus (HIV) infection: immunoultrastructural study of circulating lymphocytes  

Microsoft Academic Search

The peripheral lymphocytes of 50 cases of human immunodeficiency virus (HIV) infection (13 of acquired immune deficiency syndrome (AIDS), 17 of AIDS related complex (ARC), and 20 healthy carriers) were studied immunoultrastructurally. The prevalence of \\

W W Feremans; K Huygen; R Menu; C M Farber; J P de Caluwe; J P van Vooren; L Marcelis; L Andre; M Brasseur; H Bondue



Studies of Lymphocyte Growth and Differentiation. Progress Report, September 1, 1975--July 31, 1976.  

National Technical Information Service (NTIS)

Studies were continued on ribonuclear protein synthesis and the assembly of ribosomes in resting and stimulated lymphocytes. We demonstrated the interdependency of protein synthesis and RNA synthesis in the formation and processing of nascent ribonuclear ...

A. D. Rubin



Effect of filgrastim treatment on inflammatory cytokines and lymphocyte functions  

Microsoft Academic Search

Twenty-four healthy male volunteers received either placebo or 75, 150, or 300 ?g filgrastim (recombinant methionyl human granulocyte colony-stimulating factor) for 12 days to study effects on monocytes and lymphocytes. In all filgrastim-treated groups, tumor necrosis factor alpha (TNF-?), interleukin-12 (IL-12), and interferon gamma (IFN-?) release by whole blood in response to endotoxin (lipopolysaccharide) was reduced. IL-12 added in vitro

Thomas Hartung; Wolf-Dietrich Doecke; Daniela Bundschuh; MaryAnn Foote; Florian Gantner; Corinna Hermann; Andre Lenz; Steven Milwee; Bill Rich; Bernadett Simon; Hans-Dieter Volk; Sonja von Aulock; Albrecht Wendel



Changes of lymphocyte membrane fluidity in rheumatoid arthritis: a fluorescence polarisation study.  

PubMed Central

Fluorescence polarisation of 1,6-diphenyl-1,3,5-hexatriene was used to study the lymphocyte membrane in rheumatoid arthritis. The increase of polarisation value in the patients (n = 27) compared with healthy controls (n = 32) suggests a decrease of membrane fluidity. Moreover, erythrocyte sedimentation rate (ESR) and plasma fibrinogen concentrations were positively correlated with lymphocyte fluorescence polarisation values (r = 0.66 and r = 0.76 respectively). The results suggest that the changes in lymphocyte membrane fluidity could be involved in the pathogenetic mechanism of rheumatoid arthritis.

Beccerica, E; Piergiacomi, G; Curatola, G; Ferretti, G



Nectin-3 (CD113) Interacts with Nectin-2 (CD112) to Promote Lymphocyte Transendothelial Migration.  


Lymphocyte trafficking and migration through vascular endothelial cells (ECs) in secondary lymphoid tissues is critical for immune protection. In the present study, we investigate the role of nectin cell adhesion molecules for the migration of lymphocytes through ECs. Nectins are key players for the establishment of homotypic and heterotypic cell to cell contacts; they are required for cell to cell adherens junction formation and take part in the transendothelial migration of monocytes during the step of diapedesis, when monocytes migrate through EC junctions. We first show that Nectin-3 (CD113) is the only nectin expressed by T lymphocytes and since nectins are expressed on ECs we explored Nectin-3 potential functions in lymphocyte: EC interactions. We demonstrate that Nectin-2, expressed on ECs, is the major counter-receptor of Nectin-3. A soluble form of Nectin-3 binds to Nectin-2 localized at EC junctions and blocking Nectin-2 trans-interactions with monoclonal antibodies abolishes the binding of soluble Nectin-3 to ECs. Nectin-2 is expressed on High Endothelial venules (HEVs), where lymphocyte homing occurs in vivo. Finally, we show that Nectin-3 trans-interaction with Nectin-2 is essential for the process of lymphocyte transendothelial migration in vitro as targeting with blocking monoclonal antibodies either Nectin-3, expressed on lymphocytes, or Nectin-2, expressed on ECs, inhibits lymphocyte extravasation. The nectin family of CAMs is important for the regulation of endothelial barrier functions and transendothelial migration of immune cells. Our results demonstrate for the first time that Nectin-3 trans-interacts with Nectin-2 to promote lymphocyte and monocyte extravasation. PMID:24116228

Devilard, Elisabeth; Xerri, Luc; Dubreuil, Patrice; Lopez, Marc; Reymond, Nicolas



Nectin-3 (CD113) Interacts with Nectin-2 (CD112) to Promote Lymphocyte Transendothelial Migration  

PubMed Central

Lymphocyte trafficking and migration through vascular endothelial cells (ECs) in secondary lymphoid tissues is critical for immune protection. In the present study, we investigate the role of nectin cell adhesion molecules for the migration of lymphocytes through ECs. Nectins are key players for the establishment of homotypic and heterotypic cell to cell contacts; they are required for cell to cell adherens junction formation and take part in the transendothelial migration of monocytes during the step of diapedesis, when monocytes migrate through EC junctions. We first show that Nectin-3 (CD113) is the only nectin expressed by T lymphocytes and since nectins are expressed on ECs we explored Nectin-3 potential functions in lymphocyte: EC interactions. We demonstrate that Nectin-2, expressed on ECs, is the major counter-receptor of Nectin-3. A soluble form of Nectin-3 binds to Nectin-2 localized at EC junctions and blocking Nectin-2 trans-interactions with monoclonal antibodies abolishes the binding of soluble Nectin-3 to ECs. Nectin-2 is expressed on High Endothelial venules (HEVs), where lymphocyte homing occurs in vivo. Finally, we show that Nectin-3 trans-interaction with Nectin-2 is essential for the process of lymphocyte transendothelial migration in vitro as targeting with blocking monoclonal antibodies either Nectin-3, expressed on lymphocytes, or Nectin-2, expressed on ECs, inhibits lymphocyte extravasation. The nectin family of CAMs is important for the regulation of endothelial barrier functions and transendothelial migration of immune cells. Our results demonstrate for the first time that Nectin-3 trans-interacts with Nectin-2 to promote lymphocyte and monocyte extravasation.

Devilard, Elisabeth; Xerri, Luc; Dubreuil, Patrice; Lopez, Marc; Reymond, Nicolas



Adenoidal and tonsillar lymphocyte subsets in AIDS children.  


Tonsils and adenoids are secondary lymphoid organs exposed to the environment. The most important classifications of AIDS include the lymphocyte subsets of peripheral blood. We have studied the lymphocyte subsets in peripheral blood and secondary lymphoid organs in a control group of children suffering adenotonsillar pathology and in five children with AIDS and the same adenotonsillar pathology. The antigen surface markers were determined by flow cytometry in lymphocytes isolated from peripheral blood, and from tonsils and adenoids after tonsillectomy and adenoidectomy, in the control group and in children diagnosed with AIDS. The most important findings in tonsils and adenoids were a decrease of the total T lymphocytes, helper T lymphocytes and CD4/CD8 ratio; an increase of cytotoxic T lymphocytes and B lymphocytes, as well as a 200% increase in monocytes of AIDS-affected children. These observations show the value of analyzing the lymphocyte subsets of the tonsils and adenoids of AIDS-affected children, and establishing an earlier relation to clinical symptoms. PMID:9865438

Lopez-Gonzalez, M A; Lucas, M; Sanchez, B; Mata, F; Delgado, F



Covalently grafted BMP-7 peptide to reduce macrophage/monocyte activity: an in vitro study on cobalt chromium alloy.  


Cobalt chromium (CoCr) alloy is widely used in orthopedic implants but its functional longevity is susceptible to inflammation related complications. Reduction of the development of chronic inflammation on the biomaterial surface would enhance direct bone-implant bonding and improve implant survival and long-term results. The BMP-7 peptide was derived from the knuckle epitope of bone morphogenic protein-7 (BMP-7) and was conjugated via a cysteine amino acid at the N-terminus. Mouse RAW 264.7 monocytes/macrophages were seeded on the CoCr substrates and inflammation was induced via lipopolysaccharide (LPS) challenge. The effects of BMP-7 peptide on inflammation were evaluated by measuring the expression of inflammatory markers like toll-like receptor-4 (TLR-4), tumor necrosis factor-? (TNF-?), and monocyte chemotactic protein-1 (MCP-1). ELISA and qPCR assays were used to study the inflammatory signals. BMP-7 signaling pathway activation was shown by the presence of phosphorylation of Smad1/5/8. Utilizing the reactivity of polydopamine films to immobilize BMP-7 peptide onto metal substrates may provide a promising approach for applications in situations where reduction of inflammation around implants would be beneficial in improving surgical outcome, bone healing, and implant integration. PMID:23055400

Tan, Hark Chuan; Poh, Chye Khoon; Cai, Yanli; Soe, Min Thun; Wang, Wilson



Transfer of cholesterol from macrophages to lymphocytes in culture.  


A major feature of macrophage metabolism is its capacity to produce and export cholesterol. Several reports have shown that the manipulation of lymphocyte cholesterol content elicits important changes in lymphocyte proliferation. These findings lead to an inquiry as to whether macrophage-derived cholesterol released into the lymphocyte surroundings may be transferred to the latter thus affecting lymphocyte function. In this study, cholesterol transfer from macrophages to lymphocytes was examined in vitro using rat cells in culture. The findings indicate that there may be a significant transfer of cholesterol from [4-14C]cholesterol labeled resident peritoneal macrophages to mesenteric lymph node resting lymphocytes (up to 173.9 +/- 2.7 pmol/10(7) lymphocytes/10(7) macrophages when co-cultivated for 48 h), in a lipoprotein-dependent manner. This represents the mass transfer of ca. 17 nmoles of cholesterol molecules per 10(7) lymphocytes from 10(7) macrophages (calculated on the basis of specific radioactivity incorporated into macrophages after the pre-labelling period), which suggests that macrophages are capable of replacing the whole lymphocyte cholesterol pool every 21 h. Moreover, an 111%-increase in the total cholesterol content of lymphocytes was found after co-cultivation with macrophages for 48 h. When compared to peritoneal cells, monocytes/macrophages obtained from circulating blood leukocytes presented a much higher cholesterol transfer capacity to lymphocytes (3.06 +/- 0.10 nmol/10(7) lymphocytes/10(7) macrophages co-cultivated for 24 h). Interestingly, inflammatory macrophages dramatically reduced their cholesterol transfer ability (by up to 91%, as compared to resident macrophages). Cholesterol transfer may involve a humoral influence, since it is not only observed when cells are co-cultivated in a single-well chamber system (cells in direct contact), but also in a two-compartment system (where cells can communicate but not by direct contact). Co-cultivation with macrophages decreased the basal incorporation of [2-14C]thymidine into lymphocyte DNA and the addition of cholesterol to lymphocyte culture media also impaired the lymphocyte proliferative response to the mitogens concanavalin A (Con A) and bacterial lipopolysaccharide (LPS). The above results suggest that macrophages may transfer cholesterol to lymphocytes (from both lymph nodes and blood), thus regulating lymphocyte function by raising the intracellular cholesterol content and suppressing lymphocyte proliferative activity. If this is so, a modulatory role for the transfer of cholesterol in both physiological (e.g. immune response) and pathological conditions (e.g. atherosclerosis) may be postulated. This hypothesis is currently under investigation in our laboratory. PMID:9530517

de Bittencourt Júnior, P I; Curi, R



Upregulation of the 5-lipoxygenase pathway in human monocytes by growth and differentiation factors.  


5-Lipoxygenase (5-LO) is the key enzyme in leukotriene biosynthesis. Leukotrienes are key mediators of inflammation, allergic and innate immune reactions. 5-LO expression is mainly restricted to a variety of immune competent cells including B-lymphocytes, granulocytes and monocytes/macrophages. Here, we studied the effects of the growth or differentiation factors TGFbeta, 1,25(OH)2D3, GM-CSF and TNFalpha on 5-LO mRNA and protein expression and on 5-LO activity in human monocytes. We found that cultivation of monocytes under standard culture conditions downregulates 5-LO mRNA expression which could be prevented by addition of the four factors. Monocyte 5-LO activity was serum-dependent and cultivation of the cells in serum-free medium strongly downregulated cellular 5-LO activity which could be prevented by TGFbeta, 1,25(OH)2D3, GM-CSF and TNFalpha to different extents. The protein kinase A activator dibutyryl-cAMP blocked the effects of the four factors. The data suggest that 5-LO expression and activity in monocytes is strongly regulated by pro- and anti-inflammatory growth or differentiation factors. PMID:23923640

Wagner, M; Werz, O; Steinhilber, D



[Cytogenetic study of blood lymphocytes in children revaccinated against smallpox].  


As the result of primary re-immunization with small-pox vaccine of eight years' old children chromosome aberrations were observed in their peripheral blood lymphocytes, the frequencies being 5.3% and 7.9% on the seventh day and in a month after the reimmunization respectively. Chromosome aberrations were significantly more rare in children with a high level of immunity retained after the preceding immunization as compared to those whose immunity was weakened with time. After the elapse of 6 months following the re-immunization the frequency of chromosome aberrationd did not exceed the initial level. The changes observed in the chromosome apparatus of lymphocytes are not specific for the small-pox vaccine alone, but are the evidence of the disturbance of homeostasis of the microorganism as the result of the effect of an alien antigen. PMID:830156

Frolov, A K; Sliusarev, A A; Dement'ev, I V; Sokhin, A A; Frolov, V K



In vitro study of lymphocyte antiproliferation and cytogenetic effect by occupational formaldehyde exposure.  


Formaldehyde is the chemical substance illegally used for food preservation in meat, vegetables and fruit. A study on the antiproliferative effect and cytogenetic effect of formaldehyde on human lymphocyte was undertaken. Heparinized blood from 30 volunteers was collected and treated with formaldehyde concentrations of 0.036, 0.072, 0.15, 0.3, 0.576, 0.8 and 1.152 mg/mL, respectively, for 24 hours. Viable lymphocyte count by hemocytometer and MTT assay were carried out for detecting the antiproliferative effect of formaldehyde on human lymphocyte. Lymphocyte culture and G-banding technique were carried out for detecting the cytogenetic effect of formaldehyde. The results showed that the numbers of viable lymphocyte in the control group were 3.45 × 10(4) cells/mL. The numbers of viable lymphocyte in the experimental subgroups were 3.03 × 10( 4), 2.69 × 10(4), 2.36 × 10(4), 2.17 × 10(4), 1.92 × 10(4), 1.68 × 10(4) and 1.04 × 10(4) cells/mL, respectively, at 24 hours. The value of IC(50) was 0.92 mg/mL. The formaldehyde concentrations of 0.036, 0.072, 0.15, 0.3, 0.576, 0.8 and 1.152 mg/mL effect the lymphocyte antiproliferation (p < 0.05). Loss of chromosome was the cytogenetic effect by the formaldehyde concentration of 0.036 and 0.072 mg/mL in this study. It is concluded that formaldehyde has the antiproliferative effect and cytogenetic effect on human lymphocyte. PMID:21505003

Pongsavee, M



Circulating monocytes are reduced by sphingosine-1-phosphate receptor modulators independently of S1P3.  


Sphingosine-1-phosphate (S1P) receptors are critical for lymphocyte egress from secondary lymphoid organs, and S1P receptor modulators suppress lymphocyte circulation. However, the role of S1P receptors on monocytes is less clear. To elucidate this, we systematically evaluated monocytes in rats and mice, both in naive and inflammatory conditions, with S1P receptor modulators FTY720 and BAF312. We demonstrate that S1P receptor modulators reduce circulating monocytes in a similar time course as lymphocytes. Furthermore, total monocyte numbers were increased in the spleen and bone marrow, suggesting that S1P receptor modulation restricts egress from hematopoietic organs. Monocytes treated ex vivo with FTY720 had reduced CD40 expression and TNF-? production, suggesting a direct effect on monocyte activation. Similar reductions in protein expression and cytokine production were also found in vivo. Suppression of experimental autoimmune encephalomyelitis in mice and rats by FTY720 correlated with reduced numbers of lymphocytes and monocytes. These effects on monocytes were independent of S1P3, as treatment with BAF312, a S1P1,4,5 modulator, led to similar results. These data reveal a novel role for S1P receptors on monocytes and offer additional insights on the mechanism of action of S1P receptor modulators in disease. PMID:23436932

Lewis, Nuruddeen D; Haxhinasto, Sokol A; Anderson, Shawn M; Stefanopoulos, Dimitria E; Fogal, Steven E; Adusumalli, Prathima; Desai, Sudha N; Patnaude, Lori A; Lukas, Susan M; Ryan, Kelli R; Slavin, Anthony J; Brown, Maryanne L; Modis, Louise K



Dinitrochlorobenzene-induced colitis in the guinea-pig: studies of colonic lamina propria lymphocytes.  

PubMed Central

Dinitrochlorobenzene-induced colitis in guinea-pigs may be immunologically mediated: animals must be presensitised to dinitrochlorobenzene to develop colitis, sensitivity can be passively transferred by lymphocytes and the injury can be mitigated by immunosuppression. In this study, we examined lamina propria lymphocytes isolated from colons of animals with dinitrochlorobenzene-induced colitis, and appropriate controls. Lamina propria lymphocytes from colitis animals have a greater percentage of rabbit erythrocyte-rosetting cells (T cells) (20.1 +/- 3.0 vs 2.3 +/- 0.8, p less than .01) and a greater capacity to mediate mitogen-induced cellular cytotoxicity with phytohaemagglutinin than lamina propria lymphocytes from normal colon (% specific cytoxicity = 29.4 +/- 8.7 vs 5.0 +/- 4.5, P less than .005). There was no difference in the percentage of rosetting cells or cytotoxicity index of spleen or mesenteric lymph node lymphocytes between the colitis animals and controls. These data suggest that there are changes in the distribution and functional characteristics of lamina propria lymphocytes which correlate with mucosal cell injury in the dinitrochlorobenzene-colitis model. Images Figure

Glick, M E; Falchuk, Z M



Lymphocyte activation-31P magnetic resonance studies of energy metabolism and phospholipid pathways.  


31P NMR spectra of perfused lymphocytes embedded in alginate capsules and activated by interleukin-2 (IL-2) are remarkably different from those of control lymphocytes. The main differences are the appearance and gradual increase of phosphodiester signals, glycerophosphocholine and glycerophosphoethanolamine. These metabolic changes also occur following perfusion with phorbol ester and after incubation with phytohemagglutinin (PHA) and are not dependent on a special growth medium. Nifedipine, a calcium channel blocking drug, inhibits the effects of PHA, but not of IL-2. There are no NMR spectral differences between peripheral lymphocytes, stimulated for 3 weeks, and tumor-infiltrating lymphocytes. Thus, sustained accelerated turnover of phosphatidylcholine (PC) and phosphatidylethanolamine is an inherent feature of the activation process. 31P NMR spectra of lymphocytes are characterized by a low phosphocholine signal. Perfusion studies with high concentrations of choline and the use of dapsone, an inhibitor of phosphocholine cytidyltransferase, indicate that choline kinase plays a key role in regulating PC synthesis in human lymphocytes. PMID:8069533

Kaplan, O; Cohen, J S



Linear relationship between lymphocyte counts in peripheral blood and buffy coat collected during extracorporeal photopheresis.  


BACKGROUND: Extracorporeal photopheresis (ECP) is commonly used to treat patients with graft-versus-host disease (GVHD) and lung transplant rejection (LTR) in our institution. The quantitative relationship between the number of white blood cells treated during ECP and the cell count in peripheral blood is unclear. STUDY DESIGN AND METHODS: Patients with GVHD and LTR receiving ECP with either UVAR?XTS or CELLEX (Therakos) were prospectively recruited for this study. A complete cell count with differential was performed on preprocedural peripheral blood and samples from the collected buffy coats. Correlation analysis and linear regression were performed between cell counts in peripheral blood and buffy coat. Collection efficiency was compared between UVAR?XTS and CELLEX. RESULTS: In all 52 patients, lymphocyte counts in buffy coat and peripheral blood showed strong correlation (r values were 0.85 and 0.983 for UVAR?XTS and CELLEX, respectively; p?Monocytes also showed consistent correlation and linearity, but not neutrophils or combined white blood cells, red blood cells, or platelets. CELLEX enriched approximately twice as many lymphocytes and monocytes than UVAR?XTS per procedure (p?lymphocyte count can predict the number of lymphocytes within the buffy coat collected during ECP, which may justify the use of peripheral lymphocyte count as a surrogate for the cell dose treated per procedure. Peripheral monocyte counts may serve as an alternative. CELLEX is more efficient in collecting lymphocytes and monocytes than UVAR?XTS under conditions tested. PMID:23414109

Liu, Chang; Shah, Kalpna; Dynis, Marian; Eby, Charles S; Grossman, Brenda J



Properties of monocytes generated from haematopoietic CD34(+) stem cells from bone marrow of colon cancer patients.  


Monocytes exhibit direct and indirect antitumour activities and may be potentially useful for various forms of adoptive cellular immunotherapy of cancer. However, blood is a limited source of them. This study explored whether monocytes can be obtained from bone marrow haematopoietic CD34(+) stem cells of colon cancer patients, using previously described protocol of expansion and differentiation to monocytes of cord blood-derived CD34(+) haematopoietic progenitors. Data show that in two-step cultures, the yield of cells was increased approximately 200-fold, and among these cells, up to 60 % of CD14(+) monocytes were found. They consisted of two subpopulations: CD14(++)CD16(+) and CD14(+)CD16(-), at approximately 1:1 ratio, that differed in HLA-DR expression, being higher on the former. No differences in expression of costimulatory molecules were observed, as CD80 was not detected, while CD86 expression was comparable. These CD14(+) monocytes showed the ability to present recall antigens (PPD, Candida albicans) and neoantigens expressed on tumour cells and tumour-derived microvesicles (TMV) to autologous CD3(+) T cells isolated from the peripheral blood. Monocytes also efficiently presented the immunodominant HER-2/neu369-377 peptide (KIFGSLAFL), resulting in the generation of specific cytotoxic CD8(+) T lymphocytes (CTL). The CD14(++)CD16(+) subset exhibited enhanced cytotoxicity, though nonsignificant, towards tumour cells in vitro. These observations indicate that generation of monocytes from CD34(+) stem cells of cancer patients is feasible. To our knowledge, it is the first demonstration of such approach that may open a way to obtain autologous monocytes for alternative forms of adaptive and adoptive cellular immunotherapy of cancer. PMID:23180014

Stec, Malgorzata; Baran, Jaros?aw; Szatanek, Rafa?; Mytar, Bo?enna; Lenart, Marzena; Czupryna, Antoni; Szczepanik, Antoni; Siedlar, Maciej; Zembala, Marek



Sources of heterogeneity in human monocyte subsets  

PubMed Central

Human monocytes are commonly defined and discriminated by the extent of their cell surface expression of CD14 and CD16, with associated differences in function and phenotype related to the intensity of expression of these markers. With increasing interest into the function and behaviour of monocytes, it is important to have a clear understanding of how differing strategies of analysis can affect results and how different protocols and population backgrounds can affect this highly morphogenic cell type. Using PBMCs from populations with differing ethnicities and histories of parasite exposure we have characterized monocyte phenotype based on intensity of CD14 and CD16 expression. Using the surface markers HLA-DR, CCR2 and CX3CR1, we compared monocyte phenotype between populations and further assessed changes in monocytes with freezing and thawing of PBMCs. Our results reveal that there is a progression of surface marker expression based on intensity of CD14 or CD16 expression, stressing the importance of careful gating of monocyte subtypes. Freezing and thawing of the PBMCs has no effect generally on the monocytes, although it does lead to a decrease in CD16 and CX3CR1 expression. We show that there are differences in the monocyte populations based on ethnicity and history of exposure to the common parasites Plasmodium falciparum and Schistosoma haematobium. This study highlights that blood monocytes consist of a continuous population of cells, within which the dominant phenotype may vary dependent on the background of the study population. Comparing results from monocyte studies therefore needs to be done with great care, as ethnic background of donor population, gating strategy and processing of PBMCs may all have an effect on outcome of monocyte phenotype.

Appleby, Laura J.; Nausch, Norman; Midzi, Nicholas; Mduluza, Takafira; Allen, Judith E.; Mutapi, Francisca



M-ficolin is expressed on monocytes and is a lectin binding to N-acetyl-d-glucosamine and mediates monocyte adhesion and phagocytosis of Escherichia coli  

PubMed Central

Ficolins are a group of multimeric proteins that contain collagen-like and fibrinogen-like (FBG) sequences. Three types of ficolins have been characterized: H-, L- and M-ficolins. Both H- and L-ficolins have demonstrated lectin activities. In the present study, the FBG domain of M-ficolin was expressed and shown to bind to N-acetyl-d-glucosamine. M-ficolin mRNA was expressed in monocytes but not in the more differentiated macrophages and dendritic cells. By flow cytometry, surface biotinylation and immunoprecipitation, we showed that M-ficolin was associated with the surface of promonocytic U937 cells. M-ficolin transiently expressed in COS-7 cells was also clearly detected on the cell surface by immunoprecipitation. By flow cytometry, M-ficolin was detected on peripheral blood monocytes but not on lymphocytes or granulocytes. Immobilized rabbit anti-M-ficolin F(ab?)2 mediated U937 cell adhesion, and the antibody also inhibited phagocytosis of Escherichia coli K-12 by U937 cells. Therefore, M-ficolin might act as a phagocytic receptor or adaptor on circulating monocytes for micro-organism recognition and may potentially mediate monocyte adhesion.

Teh, C; Le, Y; Lee, S H; Lu, J



Lymphocyte traffic control by chemokines  

Microsoft Academic Search

In contrast to the remarkable chemokine responses of phagocytes and monocytes that were documented early on, lymphocytes have been considered for a long time to be poor targets for chemokine action. This view has changed dramatically with the discovery that peripheral blood T cells need to be activated before they can migrate in response to inflammatory chemo-kines. These chemokines do

Bernhard Moser; Pius Loetscher



Oxidative Mechanisms of Monocyte-Mediated Cytotoxicity  

NASA Astrophysics Data System (ADS)

Human monocytes stimulated with phorbol myristate acetate were able to rapidly destroy autologous erythrocyte targets. Monocyte-mediated cytotoxicity was related to phorbol myristate acetate concentration and monocyte number. Purified preparations of lymphocytes were incapable of mediating erythrocyte lysis in this system. The ability of phorbol myristate acetate-stimulated monocytes to lyse erythrocyte targets was markedly impaired by catalase or superoxide dismutase but not by heat-inactivated enzymes or albumin. Despite a simultaneous requirement for superoxide anion and hydrogen peroxide in the cytotoxic event, a variety of hydroxyl radical and singlet oxygen scavengers did not effect cytolysis. However, tryptophan significantly inhibited cytotoxicity. The myeloperoxidase inhibitor cyanide enhanced erythrocyte destruction, whereas azide reduced it modestly. The inability of cyanide to reduce cytotoxicity coupled with the protective effect of superoxide dismutase suggests that cytotoxicity is independent of the classic myeloperoxidase system. We conclude that monocytes, stimulated with phorbol myristate acetate, generate superoxide anion and hydrogen peroxide, which together play an integral role in this cytotoxic mechanism.

Weiss, Stephen J.; Lobuglio, Albert F.; Kessler, Howard B.



Lymphocyte microvilli are dynamic, actin-dependent structures that do not require Wiskott-Aldrich syndrome protein (WASp) for their morphology  

Microsoft Academic Search

Short microvilli cover the surfaces of circulating mammalian lymphocytes. The surfaces of monocytes and neutrophils are very different, containing ruffles as their predominant structure. In this study, we present the first quantitative character- ization of lymphocyte microvilli. From analysis of scanning electron micro- graphs, we find that median microvillar length and surface density range from 0.3 to 0.4m and 2

Sonja Majstoravich; Jinyi Zhang; Susan Nicholson-Dykstra; Stefan Linder; Wilhelm Friedrich; Katherine A. Siminovitch; Henry N. Higgs



Sex differences in human lymphocyte Na,K-ATPase as studied by labeled ouabain binding.  


Lymphocytes Na,K-ATPase is a plasma membrane enzyme that is up-regulated under lymphocytes activation. It is also studied as a model of brain cells Na,K-ATPase. Data about sex-related specificities of the enzyme are not available. The binding of tritium-labelled ouabain to lymphocyte plasma membrane Na,K-ATPase was studied in healthy volunteers of both sexes. The binding interactions were expressed in terms of K(D) and B(Max). The first parameter is related to the affinity of ouabain for the enzyme whereas the second one is related to its density on the cell membrane. Distinct sex-related differences were found. Whereas in males there is a significant direct correlation between the parameters K(D) and B(Max), in females this is not present. However, in females there is a significantly lower K(D) in the 25-37 age range. The latter result probably reflects the expression of subunit variants giving a greater affinity for ouabain. This circumstance may be relevant both to lymphocytes' ability to be activated and to brain function, if one admits that lymphocyte Na,K-ATPase faithfully represents the brain-borne one. PMID:17365113

Scarrone, S; Balestrino, M; Frassoni, F; Pozzi, S; Gandolfo, C; Podestà, M; Cupello, A



Titanates deliver metal ions to human monocytes.  


Amorphous peroxotitantes (APT) are insoluble titanium-based particles that bind a variety of metal compounds with high affinity; these particles could be sequestered locally in a solid phase to deliver metal-based drugs. Previous studies have confirmed the 'biodelivery' of metals from metal-APT complexes to fibroblasts, but not monocytes. Our goal in the current study was to use monocytic cytokine secretion to assess delivery of gold or platinum-based compounds from APT to human THP1 monocytes. Cytokine secretion was not triggered by APT alone or metal-APT complexes. In monocytes activated by lipopolysaccharide (LPS), APT alone enhanced or suppressed IL1beta or IL6 secretion, yet TNFalpha secretion was unaffected. Complexes of APT and Au(III) or cis-platin altered LPS-activated IL6 or IL1beta secretion most, TNFalpha least. Our results suggest that the APT deliver metals to monocytes. PMID:19941042

Wataha, John C; Hobbs, David T; Wong, Jacqueline J; Dogan, Sami; Zhang, Hai; Chung, K-H; Elvington, Mark C



Monocyte Adhesion Molecules Expression in Patients with Chronic Hepatitis C Liver Disease  

PubMed Central

Background Chronic viral hepatitis is histologically characterized by predominantly periportal infiltration of mononuclear cells, including lymphocytes and monocytes/macrophages. Intralobular infiltration of these inflammatory cells is an ominous sign of deterioration and a criterion for disease activity. Objective To assess the monocyte inflammatory milieu, monocytes adhesion molecules, their endothelial receptors, cytokines and chemokines in patients with HCV induced chronic liver disease, in an attempt to clarify the role of blood monocytes in induction of inflammation and fibrogenesis in chronic hepatitis C liver disease. Subjects and Methods The current study included 60 patients with chronic liver disease categorized into 2groups: Patients chronic hepatitis C (CHC) and patients with liver cirrhosis (LC), 15 patients each; 15 healthy subjects were included as normal controls. Immunophenotype characterization was carried out by flowcytometric analysis for identification of CD11a, CD11b and CD49d monocyte surface antigen expression in different groups studied. The circulating levels of the soluble adhesion molecules (sE-selectin, sICAM-1 and sVCAM-1), cytokines (TNF-? and IL-1) and chemokines (MCP-1) were also assessed by immunoassays. Results Data demonstrated a significant increase (p<0.01) in the surface expression of CD11a on peripheral blood monocytes and in the circulating levels sE-selectins, sICAM-1, sVCAM-1 and TNF-? in both groups of patients compared to healthy subjects. Data also revealed a significant increase (p<0.01) in the surface expression of each of CD11b and CD49d on peripheral blood monocytes and in the circulating levels sICAM-1, sVCAM-1 and TNF-? in patients with LC compared to those with CHC. Moreover, data demonstrated that the increase in surface antigen expression of each CD11a (p<0.01), CD11b (p<0.05) and CD49d (p<0.01) on circulating peripheral blood monocytes is positively correlated with the increase in the circulating levels of each of sICAM-1 and sVCAM-1 in the both groups of patients. Conclusions These findings suggest that the modulation of monocyte-subset recruitment into the liver via adhesion molecules or cytokines/cytokine receptors may represent promising approaches for therapeutic interventions in human liver fibrosis. Measurement of serum soluble adhesion molecules may be useful for monitoring progression of liver inflammation and fibrosis during CHC.

El-Bassiouni, Nora E.I.; Mahmoud, Ola M.; El Ahwani, Eman G; Ibrahim, Raafat A.; El Bassiouny, Azza E.I.



Interleukin 1 production during accessory cell-dependent mitogenesis of T lymphocytes  

PubMed Central

We have studied the control and significance of IL-1 production in human leukocyte cultures during accessory cell-dependent, T lymphocyte mitogenesis using sensitive bioassays and immunolabeling techniques. In primary antigen-dependent systems like the MLR, IL-1 production was not detected in accessory cells (monocytes, dendritic cells) or T cells, suggesting that it is not an early product in these responses. However, monocytes could be induced to make IL-1 after interacting with sensitized antigen-specific T cells. Both alloreactive T cell clones or freshly prepared lymphoblasts induced IL-1 provided the monocytes carried the HLA-DR antigens to which the T cells were initially sensitized. Even in these circumstances, dendritic cells and B cells failed to make IL-1. The mechanism whereby activated T cells induce IL- 1 in monocytes was explored. Supernatants from cocultures of monocytes and T cells or several recombinant cytokines induced little or no IL-1. A more potent antigen independent pathway of IL-1 induction was identified. IL-1 could be induced in third-party HLA-DR nonspecific monocytes in cocultures of alloreactive T cell clones or blasts and HLA- DR-specific dendritic cells. The induction was factor independent since dendritic cells and T blasts placed in a chamber separate from third- party monocytes by a semipermeable membrane did not induce monocyte IL- 1. These results suggest that a cell contact mechanism rather than an IL-1-inducing factor leads to IL-1 production. The role of IL-1 in T cell proliferation was tested with a polyclonal anti-IL-1 antibody. The antibody failed to block the proliferation of primary T cells, or alloreactive T cell clones and blasts stimulated with HLA-specific monocytes or dendritic cells, even though IL-1 in the medium was neutralized.



Chlamydia pneumoniae Infection Induces Differentiation of Monocytes into Macrophages  

Microsoft Academic Search

Migration and differentiation of monocytes to the intima of blood vessels may be a crucial first step in the development of atherosclerosis associated with Chlamydia (Chlamydophila) pneumoniae. However, the involve- ment of C. pneumoniae infection in such steps is not clear. In the present study, therefore, the differentiation- inducing activity of C. pneumoniae to monocytes was examined. Human THP-1 monocytic

Hiroyuki Yamaguchi; Shusaku Haranaga; Ray Widen; Herman Friedman; Yoshimasa Yamamoto



Studies on the nature of EA binding by lymphocytes from rheumatoid arthritis patients  

PubMed Central

Investigation of the nature of the increased erythrocyte-antibody (EA) binding activity of peripheral blood lymphocytes (PBL) from rheumatoid arthritis (RA) patients reported in the preceding paper has revealed that IgG is the active class of antibody in this rosette formation. Some IgM binding also occurs. SRBC sensitized with F(ab)2 preparations of IgG do not give rosette formation even at high concentrations. EA binding is inhibited by prior incubation of lymphocytes with heat-aggregated human IgG but antigen-antibody complexes did not give significant inhibition. The majority of these rosettes were found to be stable at 4°C and room temperature but labile at 37°C. Enzyme studies with pronase, trypsin, neuraminidase and treatment with sodium azide gave results strongly supporting the conclusion that the increased binding observed is increased Fc-receptor activity. This activity appears not to be a result of Fc binding by cell-bound rheumatoid factor. A range of titres of antibody and of IgG was used to sensitize erythrocytes to form EA and the enhanced EA-rosette formation by PBL from RA patients occurred throughout the range of concentrations of sensitizing antibody. Significantly more EA were bound by individual lymphocytes from RA patients than control subjects. This data suggest that the Fc receptors on RA lymphocytes are more avid for EA than receptors on lymphocytes from healthy people.

Sharpin, Rosemary K. C.; Wilson, J. D.



A quantitative ultrastructural study of peripheral blood lymphocytes containing parallel tubular arrays in Epstein-Barr virus and cytomegalovirus mononucleosis.  

PubMed Central

In the normal peripheral circulation there exists a subpopulation of lymphocytes that is ultrastructurally distinct. This lymphocyte is identified with the electron microscope by the presence of cytoplasmic microtubulelike inclusions called parallel tubular arrays (PTAs) and contains Fc-receptors for cytophilic antibody. In this study, lymphocytes containing PTAs (PTA-lymphocytes) were quantitated from serial peripheral blood specimens obtained from two patients with Epstein-Barr virus (EBV) mononucleosis and two patients with cytomegalovirus (CMV) mononucleosis. These data were then correlated with the clinical state of the patient. It was determined that both the percentage and absolute number of PTA-lymphocytes were highest during the acute phase of the illness. In follow-up specimens, three of the four patients' absolute lymphocyte count fell to within normal limits before the absolute PTA-lymphocyte count. In one patient, the absolute PTA-lymphocyte count was significantly elevated 13 months after the initial clinic visit. Although the PTA-lymphocyte count was highest during the acute phase of the illness, there was no consistent correlation with the clinical state of the patient during follow-up. The estimation of absolute PTA-lymphocyte counts was determined to be valid after a morphometric analysis of the cellular areas occupied by PTAs during the acute and convalescent phases of the disease revealed no statistical differences. Electron microscopy was also performed on the peripheral blood of a patient with syphilis. Although a hematologic workup of this patient during the acute phase of his illness revealed a large number of atypical lymphocytes, electron-microscopic examination of the same specimen revealed both a normal number and a normal percentage of PTA-lymphocytes. The immunologic role of this ultrastructurally distinct third population (non-T, non-B) of lymphocytes, or "killer cells," in the course of infectious mononucleosis is discussed. Images Figure 1 Figure 2 Figure 3 Figure 4

Payne, C. M.; Tennican, P. M.



Monocyte Chemotactic Protein 3 Is a Most Effective Basophil and Eosinophil-activating Chemokine  

Microsoft Academic Search

Summary CC chemokines constitute a novel class of cytokines that attract and activate monocytes and lymphocytes, as well as basophil and eosinophil lenkocytes, with distinct target cell profiles, and are believed to be involved in the regulation of different types of inflammation. The action of the recently identified monocyte chemotactic protein 3 (MCP-3) on human basophil and eosinophil function was

Clemens A. Dahinden; Thomas Geiser; Thomas Brunner; Daniel Caput; Pascual Ferrara; Adrian Minty; Marco BaggioliniS



[Study on T-lymphocyte culture and function of T-lymphocyte subsets in patients with aplastic anemia].  


T-lymphocytes form the peripheral blood of 12 patients with aplastic anemia (AA) and 4 patients with aplastic anaemia-paroxysmal hemoglobinuria syndrome (AA-PNH) were cultivated and their subsets were measured by monoclonal antibodies CD4, CD8, CD25 with indirect immunofluorescence technique. Change of the proportion of T-lymphocyte subsets was observed in 5 patients with AA after the mononuclear cells were activated with phytohemagglutinin (PHA). Its was shown that the number of CFU-T of the patients with AA was apparently decreased in comparison with that of normal controls (P less than 0.05). Disorder of T-lymphocyte subsets took place in patients with AA. The percentages of CD8 and CD25 lymphocytes were increased. The helper: suppressor T-lymphocytes (CD4:CD8) ratio was significantly increased (P less than 0.01). In the cases of AA-PNH, the number of CFU-T was significantly increased (P less than 0.01) and that of T-lymphocyte subsets did not change. The ratio of CD4/CD8 in 5 patients with AA was further decreased after activating mononuclear cells with PHA. These results showed that the increase of T-lymphocytes in patients with AA may account for the low level of CFU-T; the high level of CFU-T in patients with AA-PNH may be related to the multiplication of bone marrow cells. However, there may be a potential disorder of immune system on patients with AA at the same time. PMID:1831745

Fan, L H; Zhou, Y J; Wang, L F



Mouse Monocyte-Derived Chemokine Is Involved in Airway Hyperreactivity and Lung Inflammation  

Microsoft Academic Search

The cloning, expression, and function of the murine (m) homologue of human (h) monocyte-derived chemokine (MDC) is reported here. Like hMDC, mMDC is able to elicit the chemotactic migration in vitro of activated lymphocytes and monocytes. Among activated lymphocytes, Th2 cells were induced to migrate most efficiently. mMDC mRNA and protein expression is modulated during the course of an allergic

Jose-Angel Gonzalo; Yang Pan; Clare M. Lloyd; Gui-Quan Jia; Gary Yu; Barry Dussault; Christine A. Powers; Amanda E. I. Proudfoot; Anthony J. Coyle; David Gearing; Jose-Carlos Gutierrez-Ramos


Morphine modulates monocyte-macrophage conversion phase.  


Monocyte migration and their activation into the macrophage phenotype play a role in the modulation of tissue injury. We studied the effect of morphine on the monocyte-macrophage conversion phase (MMCP). Phorbol 12-myristate 13-acetate (PMA) activated THP-1 cells and promoted their adhesion to the substrate. Morphine inhibited PMA-induced MMCP. However, opiate receptor antagonists attenuated this effect of morphine. Interestingly, PMA as well as morphine-stimulated superoxide production by monocytes. Superoxide dismutase (SOD) not only inhibited PMA-mediated MMCP but also attenuated the inhibitory effect of morphine. PMA not only enhanced adhesion of monocytes to a filter but also promoted their migration. These findings suggest that the PMA-induced macrophage phenotype conversion may be accelerating their migration; whereas, morphine may be preventing the migration of monocytes by inhibiting MMCP. PMID:16698002

Hatsukari, Ikuske; Hitosugi, Naoko; Dinda, Amit; Singhal, Pravin C



Peroxisome proliferator-activated receptor and retinoid X receptor ligands inhibit monocyte chemotactic protein-1-directed migration of monocytes  

Microsoft Academic Search

Monocyte chemotactic protein-1 (MCP-1)-directed transendothelial migration of monocytes plays a key role in the development of inflammatory diseases. Infiltration of tissues by monocytes requires degradation of extracellular matrices, a process that involves matrix metalloproteinases. We studied the effects of peroxisome proliferator-activated receptor (PPAR) ?, ?, and retinoid X receptor ? (RXR?) ligands on MCP-1-directed migration and matrix metalloproteinase expression of

Ulrich Kintscher; Stephan Goetze; Shu Wakino; Sarah Kim; Sunil Nagpal; Roshantha A. S Chandraratna; Kristof Graf; Eckart Fleck; Willa A Hsueh; Ronald E Law



The use of the alkaline comet assay with lymphocytes in human biomonitoring studies  

Microsoft Academic Search

We reviewed the data of 45 alkaline comet assay studies with lymphocytes published during the last three years with the objective of monitoring human exposure to genotoxic agents as a result of occupation, drug treatment, diseases or environmental pollution. The strengths of the studies were that: (i) a lot of data could be obtained within a relatively short period of

Floriane Faust; Fekadu Kassie; Siegfried Knasmüller; Rolf Hasso Boedecker; Marion Mann; Volker Mersch-Sundermann



Monocyte subtypes predict clinical course and prognosis in human stroke.  


The number of circulating monocytes increases after stroke. In this study, we assessed the time course and phenotype of monocyte subsets and their relationship with the clinical course and outcome in 46 consecutive stroke patients and 13 age-matched controls. The proportion of the most abundant 'classical' CD14(high)CD16- monocytes did not change after stroke, whereas that of CD14(high)CD16+ monocytes increased and CD14(dim)CD16+ monocytes decreased. CD14(high)CD16+ monocytes had the highest expression of TLR2, HLA-DR and the angiogenic marker, Tie-2; CD14(dim)CD16+ monocytes had the highest expression of costimulatory CD86 and adhesion molecule CD49d. Platelet-monocyte interactions were highest in CD14(high)CD16- monocytes and lowest in CD14(dim)CD16+ monocytes. In adjusted models, 1/CD14(high)CD16- monocytes were associated with poor outcome (OR: 1.38), higher mortality (OR: 1.40) and early clinical worsening (OR: 1.29); 2/CD14(high)CD16+ monocytes were inversely related to mortality (OR: 0.32); and 3/CD14(dim)CD16+ monocytes were inversely related to poor outcome (OR: 0.74) and infarction size (r=-0.45; P=0.02). These results illustrate that the predominant monocyte subtype conveys harmful effects after stroke, which include stronger interaction with platelets. Alternatively, rarer subpopulations of monocytes are beneficial with a phenotype that could promote tissue repair and angiogenesis. Therefore, monitoring of monocyte subtypes may emerge as a useful tool at the bedside for stroke patients. PMID:19293821

Urra, Xabier; Villamor, Neus; Amaro, Sergio; Gómez-Choco, Manuel; Obach, Víctor; Oleaga, Laura; Planas, Anna M; Chamorro, Angel



2-Chlorodeoxyadenosine (cladribine) induces apoptosis in human monocyte-derived dendritic cells.  


2-Chlorodeoxyadenosine (cladribine, CdA) is an immunosuppressive drug that is licensed to treat hairy cell leukaemia, and has been shown recently to have beneficial effects in patients with multiple sclerosis (MS). The therapeutic effects of CdA have been suggested to be mediated partly through its potent toxicity towards lymphocytes. However, the effects of CdA on other immune cells are poorly understood. In the present study, we investigated the effects of CdA on the induction of apoptosis in human monocytes, monocyte-derived immature (ImDC) and mature (mDC) dendritic cells. Treatment of monocytes with CdA strongly induced apoptosis after 24 h, while apoptosis induction in DC was evident after 72 h. Furthermore, CdA treatment strongly induced caspase-3 and caspase-9 in monocytes, whereas activation of caspases was undetected in DC. The mitochondrial membrane potential in DC was reduced significantly after CdA treatment. DNA hypodiploid assessment showed fragmented nuclei in DC after CdA treatment together with activation of p53 protein. These results revealed that CdA induces caspase-independent apoptosis in DC and suggest cell type specific effects of CdA. This mechanism may contribute to the effect of CdA in autoimmune diseases. PMID:23607690

Singh, V; Prajeeth, C K; Gudi, V; Bénardais, K; Voss, E V; Stangel, M



Impaired B lymphocyte reactivity in patients after radiotherapy  

SciTech Connect

The effect of therapeutic irradiation upon B lymphocyte function was investigated in patients with various malignancies. The test system used was a reverse hemolytic plaque assay, which made it possible to study the activation and differentiation of B lymphocytes into immunoglobulin-secreting cells (ISC). Peripheral blood lymphocytes from normal individuals and patients before and after radiotherapy were stimulated in vitro with the polyclonal B cell activator pokeweed mitogen, and the number of ISC was estimated. B cell reactivity was markedly reduced in those patients who had received irradiation within the last six months. In patients in whom radiotherapy had been terminated more than 12 months before the lymphocytes were tested, B cell reactivity was comparable to that of patients prior to radiotherapy. By means of marker analyses, there was a reduction of B lymphocytes and T lymphocytes in the peripheral blood with a preponderance of T helper cells. Several mechanisms--e.g., reduced or defective B cell differentiation, altered regulatory T-helper or suppressor cell function or activation of suppressive monocytes--could be responsible for impaired B cell reactivity after radiotherapy.

Sieber, G.; Zierach, P.; Herrmann, F.; Brust, V.J.; Ruehl, H.



Safety and Tolerability Study of PCI-32765 in B Cell Lymphoma and Chronic Lymphocytic Leukemia

B-cell Chronic Lymphocytic Leukemia; Small Lymphocytic Lymphoma; Diffuse Well-differentiated Lymphocytic Lymphoma; B Cell Lymphoma; Follicular Lymphoma,; Mantle Cell Lymphoma; Non-Hodgkin's Lymphoma; Waldenstrom Macroglobulinemia; Burkitt Lymphoma; B-Cell Diffuse Lymphoma



Making a difference: Monocyte Heterogeneity in Cardiovascular Disease  

PubMed Central

Monocytes are frequently described as bone marrow-derived precursors of macrophages. Although many studies support this view, we now appreciate that monocytes neither develop exclusively in the bone marrow nor give rise to all macrophages and dendritic cells. In addition to differentiating to specific leukocyte populations, monocytes, as monocytes, are functionally and ontogenically heterogeneous. In this review we will focus on the development and activity of monocytes and their subsets in mice (Ly-6Chigh/low) and humans (CD14+/dim/? CD16+/?) in the context of atherosclerosis and its complications.

Hilgendorf, Ingo; Swirski, Filip K



Proteome Comparison of Alveolar Macrophages with Monocytes Reveals Distinct Protein Characteristics  

Microsoft Academic Search

Alveolar macrophages (AMs) are a subset of tissue macrophages situated in the alveolar milieu. Compared with their precursor blood monocytes, AMs exhibit distinct physiologic functions unique to their anatomic location. However, the molecular details that control monocyte differentiation into AMs remain unknown. This study employed a proteomic approach to define protein characteristics that distinguish AMs from monocytes. AMs and monocytes

Ming Jin; Judy M. Opalek; Clay B. Marsh; Haifeng M. Wu



Binding of fibrinogen to human monocytes.  

PubMed Central

The interaction of fibrinogen with monocytes was studied. After stimulation with ADP (10 microM) or thrombin (1 U/ml), platelet-free suspensions of human monocytes bind 125I-fibrinogen with two different affinities in a specific and Ca2+-dependent reaction with saturation at 5.80-7.35 X 10(-7) M of added protein. The binding of fibrinogen to specific receptors on monocytes induces the procoagulant activity of these cells. Thrombasthenic cells or normal monocytes preincubated with a monoclonal antibody to the platelet glycoprotein IIb/IIIa complex (10E5) do not bind fibrinogen and have no procoagulant activity. Metabolic studies with [35S]methionine revealed that cultured monocytes actually synthesize a surface antigen precipitated by 10E5 antibody as a major band with 92,000 relative molecular weight. Our data indicate that monocytes express receptors for fibrinogen only in part related to the platelet glycoprotein IIb/IIIa complex. Furthermore, the binding of fibrinogen to monocytes enhances the cooperation of these cells in hemostasis. Images

Altieri, D C; Mannucci, P M; Capitanio, A M



Cytotoxicity and Proliferation Studies with Arsenic in Established Human Cell Lines: Keratinocytes, Melanocytes, Dendritic Cells, Dermal Fibroblasts, Microvascular Endothelial Cells, Monocytes and T-Cells  

Microsoft Academic Search

Based on the hypothesis that arsenic exposure results in toxicity and mitogenecity, this study examined the dose-response of arsenic in established human cell lines of keratinocytes (HaCaT), melanocytes (1675), dendritic cells (THP-1\\/A23187), dermal fibroblasts (CRL1904), microvascular endothelial cells (HMEC), monocytes (THP- 1), and T cells (Jurkat). Cytotoxicity was determined by incubating THP-1, THP-1+ A23187 and JKT cells in RPMI 1640,

Barbara Graham-Evans; Paul B. Tchounwou; Hari H. P. Cohly



A phase I study of escalated dose subcutaneous alemtuzumab given weekly with rituximab in relapsed chronic lymphocytic leukemia/small lymphocytic lymphoma.  


This study assessed the safety and preliminary efficacy of escalated dose subcutaneous alemtuzumab in combination with rituximab in chronic lymphocytic leukemia. Twenty-eight patients with relapsed refractory chronic lymphocytic leukemia were treated on four dosing cohorts of weekly rituximab at 375 mg/m(2) and alemtuzumab doses that started at 30 mg three times per week and escalated to weekly dosing over four weeks, culminating with 90 mg weekly. One dose limiting toxicity of a rituximab infusion reaction was seen in cohort 2, but the regimen was otherwise well tolerated without evidence of differential toxicity by cohort. The overall response rate by National Cancer Institute-Working Group criteria was 61%, and the rate of complete bone marrow response was 43%, most of whom were negative for minimal residual disease. The addition of CT scan evaluation per International Workshop on Chronic Lymphocytic Leukemia 2008 criteria reduced the overall response rate to 14%. Median overall survival was 35 months, with 12 patients able to proceed to stem cell transplantation. Pharmacokinetic studies showed that chronic lymphocytic leukemia involving more than 80% of the bone marrow at study start was associated with lower trough concentrations of alemtuzumab and rituximab, and that higher trough serum concentrations of alemtuzumab were associated with complete bone marrow clearance. We conclude that escalated subcutaneous doses of alemtuzumab given weekly are well tolerated and result in excellent bone marrow clearance of chronic lymphocytic leukemia, helping patients to proceed to stem cell transplantation. This study is registered at (Identifier:00330252). PMID:23645694

Brown, Jennifer R; Messmer, Bradley; Werner, Lillian; Davids, Matthew S; Mikler, Evgeny; Supko, Jeffrey G; Fisher, David C; LaCasce, Ann S; Armand, Philippe; Jacobsen, Eric; Dalton, Virginia; Tesar, Bethany; Fernandes, Stacey M; McDonough, Sean; Ritz, Jerome; Rassenti, Laura; Kipps, Thomas J; Neuberg, Donna; Freedman, Arnold S



Toward a Refined Definition of Monocyte Subsets  

PubMed Central

In a nomenclature proposal published in 2010 monocytes were subdivided into classical and non-classical cells and in addition an intermediate monocyte subset was proposed. Over the last couple of years many studies have analyzed these intermediate cells, their characteristics have been described, and their expansion has been documented in many clinical settings. While these cells appear to be in transition from classical to non-classical monocytes and hence may not form a distinct cell population in a strict sense, their separate analysis and enumeration is warranted in health and disease.

Ziegler-Heitbrock, Loems; Hofer, Thomas P. J.



Tricyclic antidepressant-induced lipidosis in human peripheral monocytes in vitro, as well as in a monocyte-derived cell line, as monitored by spectrofluorimetry and flow cytometry after staining with Nile red.  


Human mono- and lymphocytes from peripheral blood and the monoblastoid cell line U-937 were used in this in vitro study of drug-induced lipidosis. Mono- and lymphocytes were exposed for 4 days to three different tricyclic antidepressants (TCAs), imipramine (25 microM), clomipramine (10 microM) and citalopram (80 microM). The lipophilic fluorophore Nile red, which stains intracellular lipid structures selectively, was used as a lipid probe. Fluorescence microscopy, spectrofluorimetry and flow cytometry were used to detect cellular lipidosis, as verified by electron microscopy. Our results demonstrate that imipramine, clomipramine and citalopram induce lipidosis in monocytes and U-937 cells, but not in lymphocytes. An accurate quantitation of induced intracellular lipidosis can be achieved by spectrofluorimetric and flow cytometric analysis. PMID:9260880

Xia, Z; Appelkvist, E L; DePierre, J W; Nässberger, L



Monocyte subpopulations and cardiovascular risk in chronic kidney disease.  


Chronic microinflammation and its cellular hallmark, monocyte activation, contribute substantially to the tremendous burden of cardiovascular disease (CVD) in patients with chronic kidney diseases (CKD). Monocyte heterogeneity is widely acknowledged. Cell-surface expression of CD14 and CD16 defines three functionally and phenotypically distinct subsets of monocytes: classical (CD14(++)CD16(-)) monocytes, intermediate (CD14(++)CD16(+)) monocytes, and nonclassical (CD14(+)CD16(++)) monocytes. A growing body of circumstantial evidence suggests that intermediate monocytes, in particular, contribute to the development of atherosclerosis in the general population as well as in patients with CKD. Intermediate monocytes express a unique pattern of chemokine receptors that have been implicated in atherogenesis. Moreover, this subset of monocytes is predisposed to secrete proinflammatory cytokines. Findings from epidemiological studies indicate that numbers of intermediate monocytes increase with worsening renal function, and that high cell counts predict adverse outcomes in patients undergoing dialysis as well as in patients at early stages of CKD. Based on laboratory and clinical data, intermediate monocytes are a promising therapeutic target for CVD in patients with CKD. PMID:22410492

Heine, Gunnar H; Ortiz, Alberto; Massy, Ziad A; Lindholm, Bengt; Wiecek, Andrzej; Martínez-Castelao, Alberto; Covic, Adrian; Goldsmith, David; Süleymanlar, Gültekin; London, Gérard M; Parati, Gianfranco; Sicari, Rosa; Zoccali, Carmine; Fliser, Danilo



Immunohistochemical study of monocyte chemoattractant protein-1 in the pancreas of NOD mice following cyclophosphamide administration and during spontaneous diabetes  

Microsoft Academic Search

In type 1 diabetes mellitus (T1DM), the processes which control the recruitment of immune cells into pancreatic islets are poorly defined. Complex interactions involving adhesion molecules, chemokines and chemokine receptors may facilitate this process. The chemokine, monocyte chemoattractant protein-1 (MCP-1), previously shown to be important in leukocyte trafficking in other disease systems, may be a key participant in the early

Yan Bai; Elizabeth Robinson; Ryan Chai; Jacqueline M. Ross; Shiva Reddy



Activation of lymphocytes  

PubMed Central

The technique involved in studying the activation of lymphocytes in the resting form, and their recognition as dividing and functional cells was studied, using phase contrast and agar as well as fluid culture. Standardization of technical methods was found to be essential, and the effect of variables was studied. Lymphocytes from human umbilical cord vein blood were found to be spontaneously activated. Infestation by microfilaria either diminished or entirely inhibited activation. The significance of lymphocytic cohesion was considered and the formation of colonies of activated lymphocytes on agar is described. The effects of non-human vertebrate lymphocytes and of human cells other than lymphocytes were studied. Spontaneous activation of abnormal lymphocytes was noted. Images

Pulvertaft, R. J. V.; Pulvertaft, Isobel



Study of zinc-induced changes in lymphocyte membranes using atomic force microscopy, luminescence, and light scattering methods  

NASA Astrophysics Data System (ADS)

Changes in the surface structure of lymphocyte membranes exposed to various concentrations of zinc ions are studied. It is found by atomic force microscopy that increasing the concentration of zinc ions leads to a reduction in the correlation length of the autocorrelation function of the roughness profile of a lymphocyte compared to control samples; this may indicate the existence of fine structure in the membrane surface. Fluorescence markers are used to observe a reduction in the microviscosity of the lipids in the outer monolayer of the lipid bilayer after lymphocytes are exposed to Zn ions, as well as the exposure of phosphatidylserine on the surface membrane, and the oxidation of HS-groups of membrane proteins. Calculations of the absorption coefficients of lymphocytes modified with zinc reveal the existence of absorption bands owing to the formation of metal-protein complexes and zinc oxide nanoparticles. These results indicate significant changes in the structural and functional state of lymphocyte membranes exposed to zinc ions.

Filimonenko, D. S.; Khairullina, A. Ya.; Yasinskii, V. M.; Kozlova, N. M.; Zubritskaja, G. P.; Slobozhanina, E. I.



The Italian Registry of Therapeutic Apheresis: granulocyte-monocyte apheresis in the treatment of inflammatory bowel disease. A multicentric study.  


Leukocytes are thought to play an important role in the pathogenesis of inflammatory bowel diseases; granulocyte-monocyte adsorptive (GMA) apheresis, an extracorporeal technique aimed at removing activated circulating leukocytes from the blood, may represent a safe and effective therapeutic tool in these patients. The Italian Registry of Therapeutic Apheresis performed an observational, multicentric study involving 24 Gastroenterology Units. In this study, laboratory data and clinical outcomes of 230 patients (148 males, mean age 43.5 years) affected with ulcerative colitis (UC, n = 194) or Crohn's disease (CD, n = 36) who underwent one or more cycles of GMA were analyzed. Each cycle consisted of five GMA treatments. The patients were followed up for a mean of 8.7 (min. 3 to max. 12) months. At 3 months, positive outcome was achieved in 77.7% of UC patients (72.0% remission, 5.7% clinical response) and 61.3% of CD patients (54.8% remission, 6.5% clinical response). The cumulative proportion of positive outcome at 12 months was 87.1% for UC patients (83.7% remission, 3.4% clinical response) and 77.4% for CD patients (74.2% remission, 3.2% clinical response). No single clinical or laboratory parameter among those analyzed (age, sex, disease characteristics, history of smoking, medication history, baseline values of clinical activity index (CAI)/Crohn's disease activity index (CDAI), hemoglobin, white blood cells count, and erythrocyte sedimentation rate) was independently associated with clinical outcome. The procedure was well tolerated with no significant adverse effects registered. PMID:22072543

Passalacqua, Stefano; Ferraro, Pietro Manuel; Bresci, Giampaolo; D'Ovidio, Valeria; Astegiano, Marco; Principi, Mariabeatrice; Testa, Roberto; D'Incà, Renata; Valpiani, Daniela; Armuzzi, Alessandro; Sablich, Renato; Cavallaro, Flaminia; Costa, Francesco; Di Leo, Vincenza; Colombo, Elisabetta; Santini, Alessia; Aratari, Annalisa; Lecis, Pierenrico; Saladino, Valeria; Riegler, Gabriele; Marco, Marino; Calella, Francesca; Ricci, Chiara; Guidi, Maria Luisa; Repaci, Giuseppe; Silla, Michele



Pivotal Roles of Monocytes/Macrophages in Stroke  

PubMed Central

Stroke is an important issue in public health due to its high rates both of morbidity and mortality, and high rate of disability. Hypertension, cardiovascular disease, arterial fibrillation, diabetes mellitus, smoking, and alcohol abuse are all risk factors for stroke. Clinical observations suggest that inflammation is also a direct risk factor for stroke. Patients with stroke have high levels of inflammatory cytokines in plasma, and immune cells, such as macrophages and T-lymphocytes, are noted within stroke lesions. These inflammatory events are considered as a result of stroke. However, recent studies show that plasma levels of inflammatory cytokines or soluble adhesion molecules are high in patients without stroke, and anti-inflammatory therapy is effective at reducing stroke incidence in not only animal models, but in humans as well. Statins have been shown to decrease the stroke incidence via anti-inflammatory effects that are both dependent and independent of their cholesterol-lowering effects. These reports suggest that inflammation might directly affect the onset of stroke. Microglial cells and blood-derived monocytes/macrophages play important roles in inflammation in both onset and aggravation of stroke lesions. We review the recent findings regarding the role of monocytes/macrophages in stroke.

Chiba, Tsuyoshi; Umegaki, Keizo



Studies on the Distribution of Surface Immunoglobulins on Human B-Lymphocytes  

PubMed Central

Normal human peripheral blood lymphocytes were studied with fluorescent anti-immunoglobulin antibodies and shown to have a patchy distribution of immunoglobulin on their surfaces that does not form a cap after complexing with antibody. Use of freeze-etch electron microscopy confirmed the distribution of immunoglobulin in isolated patches on the membrane. Evidence is presented that this distribution may explain the absence of capping of these human cells as compared with mouse B-lymphocytes. Studies of the metabolism of antibody bound to the cell surface revealed rapid shedding of complexes from the cell and also rapid endocytosis with subsequent degradation of the antibody. Several attempts to alter this distribution of immunoglobulin on the surface were unsuccessful. Possible mechanisms by which cell surface elements may be organized are discussed, as well as the significance of the results in terms of the immune response and the classification of certain lymphoproliferative diseases. Images

Ault, Kenneth A.; Karnovsky, Morris J.; Unanue, Emil R.



Rosiglitazone inhibits monocyte/macrophage adhesion through de novo adiponectin production in human monocytes.  


Rosiglitazone (RSG) has a variety of actions on both insulin sensitization and anti-atherogenic effects. The molecular effect of RSG on monocyte/macrophage function in terms of de novo synthesis of adiponectin is not fully understood. Here, we examined the regulation of adiponectin expression in human monocytes/macrophages by RSG and its function on monocyte adhesion during initiation of atherosclerosis. Adiponectin expression in monocytes and macrophages was studied by RT-PCR, quantitative real-time PCR, Western blot, and immunocytochemistry. Signal transduction and adhesion molecules were studied in order to describe the function of de novo synthesized adiponectin in monocyte adhesion. Adiponectin was expressed and upregulated during monocyte differentiation. The expression of adiponectin was enhanced, albeit at a much lesser degree, by a peroxisome proliferator-activated receptor gamma (PPAR gamma) agonist RSG, which was similar to what was found in adipocytes. Monocyte adhesion was remarkably reduced when the cells were treated with RSG for 12 h. This inhibitory effect of RSG was abolished by specific anti-adiponectin antibodies but not by non-immune immunoglobulin G in a serum-free condition. Adiponectin-induced suppression on monocyte adhesion was inhibited by a selective AMP-activated protein kinase (AMPK) inhibitor compound C. The reduced expression and/or function of adhesion molecule integrins may underlie the mechanism contributing to reduced monocyte adhesion upon AMPK activation. Our data suggest that the inhibitory effect of RSG on monocyte adhesion might be at least in part through de novo adiponectin expression and activation of an AMPK-dependent pathway, which might play an important role in atherogenesis. PMID:20506517

Tsai, Jaw-Shiun; Chen, Ching-Yu; Chen, Yuh-Lien; Chuang, Lee-Ming



Monocyte and macrophage heterogeneity in the heart.  


Monocytes and macrophages are innate immune cells that reside and accumulate in the healthy and injured heart. The cells and their subsets pursue distinct functions in steady-state and disease, and their tenure may range between hours and months. Some subsets are highly inflammatory, whereas others support tissue repair. This review discusses current concepts of lineage relationships and crosstalk of systems, highlights open questions, and describes tools for studying monocyte and macrophage subsets in the murine and human heart. PMID:23743228

Nahrendorf, Matthias; Swirski, Filip K



Interference study on the free calcium in the human peripheral lymphocyte by acrylamide  

Microsoft Academic Search

The role of acrylamide on the human peripheral lymphocytes was studied by laser scanning confocal microscopy technique and fluo-3. The calibration value of the apparent dissociation constant (Kd) of the fluo-3-Ca2+ complex was obtained as 4.83×10?7mol\\/L. Acrylamide (<54?g\\/mL) evoked a rise in free intracellular calcium concentration [Ca2+]i, in a dose-dependent manner. Acrylamide induced the increase of [Ca2+]i was discussed in

Hai Yan Wang; Chun Gui Zhao; Ye Hong Zhou; Shao Min Shuang; Chuan Dong



Serotonin transporter is differentially localized in subpopulations of lymphocytes of major depression patients. Effect of fluoxetine on proliferation.  


Modifications of lymphocyte serotonergic system have been described in major depression. The aim of this study was to determine new possible changes of this system in depression. Twenty eight patients, free of drugs, diagnosed with major depression disorder by Structured Clinical Interview for Disorders of Axis I, without medical illnesses, written consent, approved by Ethical Committees were included. Controls were 30 healthy subjects without family history of psychiatric disease. Blood monocytes were isolated with Ficoll/Hypaque, and lymphocytes by differential adhesion to plastic. Serotonin and 5-hydroxyindoleacetic acid determined by HPLC. Monocytes had higher serotonin concentrations than lymphocytes, and serotonin/5-hydroxyindoleacetic acid was lower in patients. Basal proliferation was elevated in depressed and not increased by Concanavalin A. Fluoxetine reduced basal proliferation more efficiently in patients, indicating activation of lymphocytes in depression. The number of cells expressing serotonin transporter was reduced in depressed. There were no differences in CD4+ (approximately 50%) or CD8+ (approximately 25%) lymphocytes between the groups, although CD8+ were lower in depressed, and greater number of them co-localized serotonin transporter than CD4+, which could be crucial for function in relation to serotonin and its receptors in immune cells. Lymphocytes were activated in this group of patients and fluoxetine reduced proliferation, probably being relevant for the psychopharmacological treatment of depression. PMID:18462811

Fazzino, Fili; Montes, Carol; Urbina, Mary; Carreira, Isabel; Lima, Lucimey



Engagement of Siglec-7 receptor induces a pro-inflammatory response selectively in monocytes.  


Sialic acid binding immunoglobulin-like lectin-7 (Siglec-7) is a trans-membrane receptor carrying immunoreceptor tyrosine based inhibitory motifs (ITIMs) and delivering inhibitory signals upon ligation with sialylated glycans. This inhibitory function can be also targeted by several pathogens that have evolved to express sialic acids on their surface to escape host immune responses. Here, we demonstrate that cross-linking of Siglec-7 by a specific monoclonal antibody (mAb) induces a remarkably high production of IL-6, IL-1?, CCL4/MIP-1?, IL-8 and TNF-?. Among the three immune cell subsets known to constitutively express Siglec-7, the production of these pro-inflammatory cytokines and chemokines selectively occurs in monocytes and not in Natural Killer or T lymphocytes. This Siglec-7-mediated activating function is associated with the phosphorylation of the extracellular signal-regulated kinase (ERK) pathway. The present study also shows that sialic acid-free Zymosan yeast particles are able to bind Siglec-7 on monocytes and that this interaction mimics the ability of the anti Siglec-7 mAb to induce the production of pro-inflammatory mediators. Indeed, blocking or silencing Siglec-7 in primary monocytes greatly reduced the production of inflammatory cytokines and chemokines in response to Zymosan, thus confirming that Siglec-7 participates in generating a monocyte-mediated inflammatory outcome following pathogen recognition. The presence of an activating form of Siglec-7 in monocytes provides the host with a new and alternative mechanism to encounter pathogens not expressing sialylated glycans. PMID:23029261

Varchetta, Stefania; Brunetta, Enrico; Roberto, Alessandra; Mikulak, Joanna; Hudspeth, Kelly L; Mondelli, Mario U; Mavilio, Domenico



Immunologic studies in two patients with persistent lymphocytic thyroiditis, thyrotoxicosis, and low radioactive iodine uptake  

SciTech Connect

Two patients with persistent lymphocytic thyroiditis and thyrotoxicosis were studied. Both patients presented with severe hyperthyroidism of nine months' duration and had nontender, small thyroid glands. Uptake of radioactive iodine (131I) was consistently low. Serum thyroxine and triiodothyronine levels remained elevated without remission until thyroidectomy. The serum thyroglobulin level was normal, but testing for microsomal antibody gave weakly positive results in one case. Thyroglobulin and thyroid stimulatory antibodies were not found. The ratio of helper to suppressor T cells was elevated in one case. Neither patient showed response to propranolol, prednisone, or iodine. Light microscopic and immunohistologic studies showed severe lymphocytic thyroiditis with formation of secondary lymphoid follicles. Lymphocytes were predominately T cells (OKT11-positive), primarily helper/inducer T cells (OKT4-positive). Hyperplastic nodules contained high immunoreactive thyroglobulin and thyroxine levels. Aberrant thymus was seen within the thyroid. These studies suggest the possibility of intrathyroidal stimulation and hydrolysis of thyroglobulin within thyroid cells and also support the hypothesis that T and B cell immunoregulatory defects are important in the pathogenesis of this disease.

Elliot, I.; Gupta, M.; Hostetter, A.; Sheeler, L.; Skillern, P.; Tubbs, R.



Immunogenetics of CD4 lymphocyte count recovery during antiretroviral therapy: An AIDS Clinical Trials Group study.  


During antiretroviral therapy, CD4 lymphocyte count increases are modest in some patients despite virologic control. We explored whether polymorphisms in genes important for T cell expansion, survival, and apoptosis are associated with the magnitude of CD4 lymphocyte count recovery during antiretroviral therapy. We studied treatment-naive individuals who achieved sustained control of plasma viremia (<400 HIV-1 RNA copies/mL) for at least 48 weeks after initiation of antiretroviral therapy and compared genotypes among individuals who had an increase of either <200 or > or =200 CD4 cells/mm3 from baseline. A total of 137 single-nucleotide polymorphisms across 17 genes were characterized in 873 study participants. In multivariate analyses that controlled for clinical variables, polymorphisms in genes encoding tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), TNF- alpha , Bcl-2-interacting molecule (Bim), interleukin (IL)-15, and IL-15 receptor alpha chain (IL-15R alpha ) were associated with the magnitude of the increase in CD4 lymphocyte count, as were haplotypes in genes encoding interferon- alpha , IL-2, and IL-15R alpha (P < .05, for each). Multifactor dimensionality reduction identified a gene-gene interaction between IL-2/IL-15 receptor common beta chain and IL-2/IL-7/IL-15 receptor common gamma chain. Immune recovery during antiretroviral therapy is a complex phenotype that is influenced by multiple genetic variants. Future studies should validate these tentative associations and define underlying mechanisms. PMID:16991084

Haas, David W; Geraghty, Daniel E; Andersen, Janet; Mar, Jessica; Motsinger, Alison A; D'Aquila, Richard T; Unutmaz, Derya; Benson, Constance A; Ritchie, Marylyn D; Landay, Alan



In-111 tropolone complex for study of lymphocyte kinetics: Evidence for an induced defect in structure, function and viability  

SciTech Connect

The lipid soluble In-111 and tropolone complex (In-T) has been proposed as a desirable cell labeling moiety for in vivo studies. Its advantages over In-111 complexed to oxy/sup -/ or acetylacetonate are water solubility and efficient cell labeling in plasma. The authors examined the effect of In-T on lymphocyte integrity and function in preparation for studies of lymphocyte kinetics in traffic. At equal concentrations, both normal and lymphocytes from patients with chronic lymphocytic leukemia had cellular In-T uptake consistently 20% greater than that achieved with In-111 oxine. This desirable uptake led to studies of function and viability. Lymphocyte mitogenmediated blastogenic capability (an intrinsic lymphocyte function) was measured in vitro in ficoll-hypaque isolated normal lymphocytes with varying concentrations and intervals of exposure of In-T. Marked impairment of lymphocyte blastogenic responsiveness was seen with 3 different mitogens (concanavalin A, phytohemmagglutinin P, and pokeweed mitogen). Severe functional impairment was seen when cells were exposed to a In-T concentration of 10 for 20 minutes; and a lesser effect was noted even at 10-minute incubation exposure. Cell viability, evaluated by trypan blue exclusion, was normal immediately following cell labeling, but rapidly and progressively failed to exclude (i.e. effective viability). Scanning electron microscopy demonstrated loss of the normal surface villous architecture within 36 hours of in vitro incubation following a 20-minute exposure. Thus, although In-T has attractive features, its effect on lymphocyte structure, function and viability eliminate it for in vivo studies in traffic kinetics.

Balaban, E.; Simon, T.R.; Kulkarni, P.; White, J.; Newton, M.; Frenkel, E.



Evidence for specific annexin I-binding proteins on human monocytes.  

PubMed Central

Recombinant human annexin I and a monoclonal antibody specific for this protein (mAb 1B) were used to investigate surface binding of this member of the annexin family of proteins to peripheral blood monocytes. Flow cytometric analysis demonstrated trypsin-sensitive, saturable binding of annexin I to human peripheral blood monocytes but not to admixed lymphocytes. A monoclonal antibody that blocks the anti-phospholipase activity of annexin I also blocked its binding to monocytes. These findings suggest the presence of specific binding sites on monocytes. Furthermore, surface iodination, immunoprecipitation and SDS/PAGE analysis were used to identify two annexin I-binding proteins on the surface of monocytes with molecular masses of 15 kDa and 18 kDa respectively. The identification and characterization of these annexin I-binding molecules should help us to better understand the specific interactions of annexin I with monocytes that lead to down-regulation of pro-inflammatory cell functions.

Goulding, N J; Pan, L; Wardwell, K; Guyre, V C; Guyre, P M



Effect of Hyperglycemia on Human Monocyte Activation  

PubMed Central

Our recent study defined the chemokine induced human monocyte signaling under normoglycemic condition. To explore the hyperglycemia induced monocyte signaling, we performed adhesion, migration and transmigration assays on human monocytes obtained from THP-1 cell line in the presence of normal (5mM) and high (10 and 20mM) glucose concentrations without chemokines. We observed augmented (p<0.01) monocyte adhesion to HUVEC monolayer at 10mM than 5mM glucose with no further increase at 20mM glucose concentration (p<0.03 vs. 10mM; p<0.01 vs. 5mM). But incremental increase in monocyte migration (p<0.01), transmigration (p<0.01) and stress fibre response (p<0.01) were observed at 10 and 20mM glucose concentrations in comparison to 5mM glucose concentrations. We found gradational increase (p<0.01) in phosphorylation of Akt S473 and glycogen synthase kinase (GSK3? S9) in hyperglycemia (10 and 20mM) when compared to 5mM glucose. Furthermore hyperglycemia (both 10 mM and 20 mM) treated monocyte showed upregulated phosphorylation of p101 and p110? subunits of PI-3 kinase in comparison to 5mM glucose. Hyperglycemia induced monocyte migration was restored to basal levels in the presence of PI-3 kinase inhibitor, LY. These observations imply that modest hyperglycemia per se, as is commonly observed in diabetic individuals, is a potent stimulator of monocyte activity even without chemokines.

Nandy, Debashis; Janardhanan, Rajiv; Mukhopadhyay, Debabrata; Basu, Ananda



A study on binding of suspended nodal lymphocytes to high endothelial venules in sections of frozen rat lymph nodes.  

PubMed Central

Live lymphocytes were previously shown to bind selectively to endothelial cells (HEV cells) of the high endothelial venules (HEVs) in sections of frozen lymph nodes. This study examines aspects of the assay that have so far not been considered or which have been incompletely addressed. It was found that bound lymphocytes form a cytoplasmic veil, and that about half of them form small groups in which they are linked together by cytoplasmic bridges. It was also found that at least 83% of the lymphocytes bind to HEV walls, but very unevenly, and that 5% also bind to medullary venules. In addition, 31% of the lymphocytes were estimated to bind to the abluminal face of HEV cells and probably to tissue lymphocytes present in subendothelial spaces or in perivascular channels (spaces). This would reflect cell interactions occurring, in vivo, between HEV cells and/or subendothelial lymphocytes. It is suggested that antigen specificity of lymphocyte retention by HEV cells accounts for the uneven binding to HEVs. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 Fig. 7 Fig. 8 Fig. 9 Fig. 10 Fig. 11 Fig. 12 Fig. 13 Fig. 14 Fig. 15 Fig. 17 Fig. 18 Fig. 19 Fig. 20 Fig. 21 Fig. 22 Fig. 23

Sainte-Marie, G; Peng, F S



The anti-idiotypic antibody 1F7 stimulates monocyte interleukin-10 production and induces endotoxin tolerance  

PubMed Central

Background Pathogens that establish chronic infection elicit immune responses with suppressive cytokines dominating over pro-inflammatory cytokines. Chronic hepatitis C virus (HCV) infection, human immunodeficiency virus (HIV) infection and simian immunodeficiency virus (SIV) infection are associated with high levels of antiviral antibodies expressing a common idiotype specifically recognized by the 1F7 monoclonal antibody (mAb). The 1F7 mAb is a murine IgM? antibody raised against immunoglobulin pooled from the plasma of multiple HIV-infected individuals. In this study, we investigated direct effects of the 1F7 mAb itself on peripheral blood mononuclear cells (PBMC). Methods Isolated monocytes or PBMC from healthy controls were incubated with the 1F7 mAb or IgM? mAb control. Cytokine production was measured in cell culture supernatants by ELISA and cells producing interleukin-10 (IL-10) were identified by subset depletion and intracellular flow cytometry. Endotoxin tolerance was assessed by exposing monocytes to lipopolysaccharide (LPS) following 1F7 mAb or IgM? mAb control pre-treatment and comparing tumor necrosis factor (TNF)-? levels in cell culture supernatants. Results The 1F7 mAb stimulated monocytes and CD36+ lymphocytes to produce IL-10 in a time and dose-dependent manner. Treatment of monocytes with 1F7 mAb also reduced their subsequent responsiveness to LPS stimulation. Conclusions Induction of antibodies expressing the 1F7 idiotype by chronic pathogens may facilitate IL-10 production and progression to chronic infection. Direct effects of IL-10 from human monocytes stimulated by 1F7-like antibodies, followed by monocyte transition to an alternatively activated phenotype illustrated by endotoxin tolerance, are two complementary features favouring a tolerogenic or non-responsive immunological environment.



Dysferlin regulates cell adhesion in human monocytes.  


Dysferlin is mutated in a group of muscular dystrophies commonly referred to as dysferlinopathies. It is highly expressed in skeletal muscle, where it is important for sarcolemmal maintenance. Recent studies show that dysferlin is also expressed in monocytes. Moreover, muscle of dysferlinopathy patients is characterized by massive immune cell infiltrates, and dysferlin-negative monocytes were shown to be more aggressive and phagocytose more particles. This suggests that dysferlin deregulation in monocytes might contribute to disease progression, but the molecular mechanism is unclear. Here we show that dysferlin expression is increased with differentiation in human monocytes and the THP1 monocyte cell model. Freshly isolated monocytes of dysferlinopathy patients show deregulated expression of fibronectin and fibronectin-binding integrins, which is recapitulated by transient knockdown of dysferlin in THP1 cells. Dysferlin forms a protein complex with these integrins at the cell membrane, and its depletion impairs cell adhesion. Moreover, patient macrophages show altered adhesion and motility. These findings suggest that dysferlin is involved in regulating cellular interactions and provide new insight into dysferlin function in inflammatory cells. PMID:23558685

de Morrée, Antoine; Flix, Bàrbara; Bagaric, Ivana; Wang, Jun; van den Boogaard, Marlinde; Grand Moursel, Laure; Frants, Rune R; Illa, Isabel; Gallardo, Eduard; Toes, Rene; van der Maarel, Silvère M



Haemolytic activity of human blood monocytes. Lysis of human erythrocytes treated with anti-A serum  

PubMed Central

Purified monocytes and lymphocytes from peripheral blood of healthy human donors were tested in vitro for cytotoxicity against blood group A erythrocytes (RBC) treated with a human hyperimmune anti-A serum. Haemolysis was quantitated by the release of radioactivity from RBC pre-labelled with 51Cr-chromate. The antiserum contained high titre antibodies which agglutinated A-RBC. After separation of serum on a Sephadex G-200 column the IgG containing fraction agglutinated A-RBC and precipitated blood group A substance. No or only weak antibody activity was detected in the IgA- and IgM-containing fractions. Purified monocytes added to a 100-fold excess of RBC in the presence of 0·1% antiserum induced some haemolysis. It was calculated that one monocyte was able to lyse 2–3 RBC within 18 hr incubation. In contrast, lymphocyte suspensions containing more than 97% small lymphocytes had no or only weak haemolytic activity at a lymphocyte-RBC ratio of 25: 1. The effector cells of the monocyte fraction adhered to glass and were eliminated by incubation with carbonyl iron. Phagocytosis by monocytes of antibody-treated RBC was frequently observed. Loading of monocytes by treatment with heat-killed Candida albicans or carbonyl iron particles suppressed their haemolytic action. Cytotoxicity was impaired after treatment of monocytes with 10?4 M sodium iodo acetate. After separation of serum on Sephadex G-200 column all monocyte induced haemolytic activity was found in the IgG containing fraction. It is assumed that haemolysis is induced by the interaction of monocytes with an IgG anti-A antibody of the antiserum. ImagesFig. 1

Holm, G.; Hammarstrom, S.



Anti-Leukocyte Serum Procurement and Population Genetic Studies of Lymphocyte Typing in Hawaii.  

National Technical Information Service (NTIS)

An attempt was made to further develope the procurement of human anti-lymphocyte serum containing monospecificity or duospecificity, obtained from pregnant women in the State of Hawaii. In addition to the above task, lymphocyte typing, of the population i...

M. Yokoyama



Syzygium cumini (Jamun) reduces the radiation-induced DNA damage in the cultured human peripheral blood lymphocytes: a preliminary study  

Microsoft Academic Search

The effects of various concentrations (0.0, 1.56, 3.125, 6.25, 12.5, 25, 50 and 100 ?g\\/ml) of the leaf extract of Syzygium cumini Linn. or Eugenia cumini (SC; black plum, Jamun, family Myrtaceae) was studied on the alteration in the radiation-induced micronuclei formation in the cultured human peripheral blood lymphocytes. Treatment of lymphocytes to various concentrations of SC resulted in a

Ganesh Chandra Jagetia; Manjeshwar Shrinath Baliga



Human Lymphocyte Apoptosis after Exposure to Influenza A Virus  

PubMed Central

Infection of humans with influenza A virus (IAV) results in a severe transient leukopenia. The goal of these studies was to analyze possible mechanisms behind this IAV-induced leukopenia with emphasis on the potential induction of apoptosis of lymphocytes by the virus. Analysis of lymphocyte subpopulations after exposure to IAV showed that a portion of CD3+, CD4+, CD8+, and CD19+ lymphocytes became apoptotic (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling positive). The percentage of cells that are infected was shown to be less than the percentage of apoptotic cells, suggesting that direct effects of cell infection by the virus cannot account fully for the high level of cell death. Removal of monocytes-macrophages after IAV exposure reduced the percent of lymphocytes that were apoptotic. Treatment of virus-exposed cultures with anti-tumor necrosis factor alpha did not reduce the percentage of lymphocytes that were apoptotic. In virus-exposed cultures treated with anti-FasL antibody, recombinant soluble human Fas, Ac-DEVD-CHO (caspase-3 inhibitor), or Z-VAD-FMK (general caspase inhibitor), apoptosis and production of the active form of caspase-3 was reduced. The apoptotic cells were Fas-high-density cells while the nonapoptotic cells expressed a low density of Fas. The present studies showed that Fas-FasL signaling plays a major role in the induction of apoptosis in lymphocytes after exposure to IAV. Since the host response to influenza virus commonly results in recovery from the infection, with residual disease uncommon, lymphocyte apoptosis likely represents a part of an overall beneficial immune response but could be a possible mechanism of disease pathogenesis.

Nichols, Joan E.; Niles, Jean A.; Roberts, Norbert J.



Human Brucellosis Is Characterized by an Intense Th1 Profile Associated with a Defective Monocyte Function?  

PubMed Central

In animal models, a defective Th1 response appears to be critical in the pathogenesis of brucellosis, but the Th1 response in human brucellosis patients remains partially undefined. Peripheral blood from 24 brucellosis patients was studied before and 45 days after antibiotherapy. Twenty-four sex- and age-matched healthy donors were analyzed in parallel. Significantly increased levels of interleukin 1? (IL-1?), IL-2, IL-4, IL-6, IL-12p40, gamma interferon (IFN-?), and tumor necrosis factor alpha (TNF-?), but not of IL-10, in serum and/or significantly increased percentages of samples with detectable levels of these cytokines, measured by enzyme-linked immunosorbent assays (ELISA), were found for untreated brucellosis patients, but these levels were reduced and/or normalized after treatment. Flow cytometry studies showed that the intracytoplasmic expression of IFN-?, IL-2, and TNF-?, but not that of IL-4, by phorbol myristate-activated CD4+ CD3+ and CD8+ CD3+ T lymphocytes was significantly increased in untreated brucellosis patients and was also partially normalized after antibiotherapy. The percentage of phagocytic cells, the mean phagocytic activity per cell, and the phagocytic indices for monocytes at baseline were defective and had only partially reverted at follow-up. T lymphocytes from untreated brucellosis patients are activated in vivo and show Th1 cytokine production polarization, with strikingly high serum IFN-? levels. In spite of this Th1 environment, we found deficient effector phagocytic activity in peripheral blood monocytes.

Rodriguez-Zapata, Manuel; Matias, Marlene J.; Prieto, Alfredo; Jonde, Marco A.; Monserrat, Jorge; Sanchez, Lorenzo; Reyes, Eduardo; De la Hera, Antonio; Alvarez-Mon, Melchor



Lymphocytic gastritis: a newly described entity: a retrospective endoscopic and histological study  

Microsoft Academic Search

Lymphocytic gastritis is a histopathological entity characterised by the accumulation of small lymphocytes in the surface and foveolar epithelium. In order to investigate the correlation between endoscopy and histology in this condition, 192 observations selected on the basis of a presumed diagnosis of erosive or varioliform gastritis were reviewed. Ninety two instances corresponded to lymphocytic gastritis, while 100 did not

J Haot; L Hamichi; L Wallez; P Mainguet



Ebola Virus Exploits a Monocyte Differentiation Program To Promote Its Entry  

PubMed Central

Antigen-presenting cells (APCs) are critical targets of Ebola virus (EBOV) infection in vivo. However, the susceptibility of monocytes to infection is controversial. Studies indicate productive monocyte infection, and yet monocytes are also reported to be resistant to EBOV GP-mediated entry. In contrast, monocyte-derived macrophages and dendritic cells are permissive for both EBOV entry and replication. Here, freshly isolated monocytes are demonstrated to indeed be refractory to EBOV entry. However, EBOV binds monocytes, and delayed entry occurs during monocyte differentiation. Cultured monocytes spontaneously downregulate the expression of viral entry restriction factors such as interferon-inducible transmembrane proteins, while upregulating the expression of critical EBOV entry factors cathepsin B and NPC1. Moreover, these processes are accelerated by EBOV infection. Finally, ectopic expression of NPC1 is sufficient to rescue entry into an undifferentiated, normally nonpermissive monocytic cell line. These results define the molecular basis for infection of APCs and suggest means to limit APC infection.

Martinez, Osvaldo; Johnson, Joshua C.; Honko, Anna; Yen, Benjamin; Shabman, Reed S.; Hensley, Lisa E.; Olinger, Gene G.



Monocytes Control Second-Phase Neutrophil Emigration in Established Lipopolysaccharide-induced Murine Lung Injury  

PubMed Central

Rationale: Acute lung injury (ALI) is an important cause of morbidity and mortality, with no currently effective pharmacological therapies. Neutrophils have been specifically implicated in the pathogenesis of ALI, and there has been significant research into the mechanisms of early neutrophil recruitment, but those controlling the later phases of neutrophil emigration that characterize disease are poorly understood. Objectives: To determine the influence of peripheral blood monocytes (PBMs) in established ALI. Methods: In a murine model of LPS-induced ALI, three separate models of conditional monocyte ablation were used: systemic liposomal clodronate (sLC), inducible depletion using CD11b diphtheria toxin receptor (CD11b DTR) transgenic mice, and antibody-dependent ablation of CCR2hi monocytes. Measurements and Main Results: PBMs play a critical role in regulating neutrophil emigration in established murine LPS-induced lung injury. Gr1hi and Gr1lo PBM subpopulations contribute to this process. PBM depletion is associated with a significant reduction in measures of lung injury. The specificity of PBM depletion was demonstrated by replenishment studies in which the effects were reversed by systemic PBM infusion but not by systemic or local pulmonary infusion of mature macrophages or lymphocytes. Conclusions: These results suggest that PBMs, or the mechanisms by which they influence pulmonary neutrophil emigration, could represent therapeutic targets in established ALI.

Scholefield, Emma; Ferenbach, David; Gibbons, Michael; Duffin, Rodger; Dorward, David A.; Morris, Andrew Conway; Humphries, Duncan; MacKinnon, Alison; Wilkinson, Tom S.; Wallace, William A. H.; van Rooijen, Nico; Mack, Matthias; Rossi, Adriano G.; Davidson, Donald J.; Hirani, Nik; Hughes, Jeremy; Haslett, Chris; Simpson, A. John



Phagocytic and Bactericidal Properties of Normal Human Monocytes  

PubMed Central

The bactericidal and phagocytic capacities of monocytes for E. coli, Staphylococcus, Salmonella, and Listeria, and factors that influence these functions were evaluated and compared with those of the polymorphonuclear leukocytes of 30 normal human subjects. Monocytes killed a significantly smaller proportion of each of the bacterial species than did neutrophils from the same individuals. Whereas the neutrophils of all individuals demonstrated the ability to kill significant numbers of the four bacterial species, there was a marked variation in the effect of monocytes of different individuals on the growth curves of these same bacteria. When the bactericidal capacity of an individual's monocytes to more than one species of bacteria was examined in the same experiment, a significant difference in the effect of monocytes on the growth curve of one bacterial species as opposed to another was noted in 4 of 17 subjects. The bactericidal ability of monocytes of single individuals was consistent on different days in 9 of the 11 subjects whose monocytes were examined more than once against the same bacteria. Studies were performed to determine if the lesser bactericidal capability of monocytes was due to a difference in the ability of monocytes and neutrophils to phagocytize or to a difference in the ability of these cells to kill ingested bacteria or both. The results demonstrated that monocytes phagocytize bacteria significantly less well than neutrophils, but the intracellular killing capacity of both cell types is equal. Addition of phenylbutazone to cell suspensions completely inhibited intracellular killing by both monocytes and neutrophils, suggesting the possibility that the bactericidal mechanisms in both cell types might be similar. Monocyte killing of E. coli, Salmonella, and Listeria, but not of Staphylococcus, was significantly diminished in heat-inactivated autologous serum. Neither increasing the concentration of autologous serum from 10% to 25% nor replacement of autologous serum with pooled human serum had any effect on monocyte killing of any of the four bacteria. These studies demonstrate that peripheral blood monocytes are less bactericidal for the four bacterial species than neutrophils, solely because monocytes are less phagocytic. A baseline for further study of factors that influence monocyte function and for study of this cell in selected patient populations is provided. Images

Steigbigel, Roy T.; Lambert, Lewis H.; Remington, Jack S.



Role of CCR8 and Other Chemokine Pathways in the Migration of Monocyte-derived Dendritic Cells to Lymph Nodes  

Microsoft Academic Search

Studying the influence of chemokine receptors (CCRs) on monocyte fate may reveal informa- tion about which subpopulations of monocytes convert to dendritic cells (DCs) and the migration pathways that they use. First, we examined whether prominent CCRs on different monocyte subsets, CCR2 or CX 3 CR1, mediated migration events upstream of the accumulation of monocyte-derived DCs in lymph nodes (LNs).

Chunfeng Qu; Emmerson W. Edwards; Frank Tacke; Véronique Angeli; Jaime Llodrá; Guzman Sanchez-Schmitz; Alexandre Garin; Nasreen S. Haque; Wendy Peters; Nico van Rooijen; Carmen Sanchez-Torres; Jonathan Bromberg; Israel F. Charo; Steffen Jung; Sergio A. Lira; Gwendalyn J. Randolph



Glucocorticoid suppression of human lymphocyte DNA synthesis. Influence of phytohemagglutinin concentration  

SciTech Connect

Glucocorticoids have been shown to suppress lectin-stimulated lymphocyte DNA synthesis in some studies, whereas in other studies, the hormones have had little effect. We have found that the position on the PHA dose-response curve that is studied is the most important determinant of whether cortisol inhibits /sup 3/H-thymidine incorporation into lymphocyte DNA. The proportion of monocytes in culture also influenced the cortisol effect, but it was quantitatively less important than PHA concentration. Cortisol (5 nM to 100 had little effect on blastogenesis or thymidine incorporation into DNA in cultures that contained both a high concentration (14% +- 2 (S.E.)) of monocytes and a concentration of PHA (0.6 to 1.2 that produced maximal stimulation of mitogenesis. When monocytes were reduced from 14 to 1.4%, cortisol (5 caused a 30% reduction in thymidine incorporation in cultures stimulated by 0.6 to 1.2 PHA. Much greater cortisol suppression of thymidine incorporation occurred if the concentration of PHA was reduced. For example, reduction of the PHA concentration from 1.2 to 0.075 resulted in an increase in suppression by 5 cortisol from 5 to 90% even in the presence of 14% monocytes. These data indicate that the suppressive effects of glucocorticoids on blastogenesis and thymidine incorporation in vitro depend principally on the concentration of PHA used to stimulate blastogenesis and secondarily on the proportion of monocytes in the culture system.

Segel, G.B.; Lukacher, A.; Gordon, B.R.; Lichtman, M.A.



Impaired in vitro survival of monocytes from patients with HIV infection.  

PubMed Central

In vitro survival of monocytes (MO) was studied in 59 patients with HIV infection of different clinical stages. MO from 61 donors and 12 healthy seronegative homosexual men were also examined. Compared with the number of MO seeded, the percentage of adherent monocyte-derived macrophages (MDM) present after 10 days was significantly lower in patients with HIV infection than in the controls. However, the number of viable, non-adherent MO/MDM was similar in patients and controls. Our data indicate markedly decreased in vitro survival of MO from patients with HIV infection. After 10 days, the MDM population in the patient cultures was significantly less differentiated than the control cells, assessed by immunocytochemical staining with monoclonal antibodies against differentiation antigens. Reduced in vitro survival of MO/MDM was associated with low numbers of CD4+ and CD8+ lymphocytes in blood, reduced lymphocyte mitogen responses, presence of HIV p24 antigen in serum and advanced clinical stage. Decreased in vitro survival of MO/MDM may be associated with HIV replication in the cells. Although the level of HIV replication in the cultures was low as assessed by measurement of HIV p24 antigen in culture supernatants and staining of MO/MDM for HIV antigens, cytopathogenic effects of HIV or HIV products cannot be ruled out. Images Fig. 1

Muller, F; Rollag, H; Gaudernack, G; Fr?land, S S



Productive human immunodeficiency virus type 1 (HIV-1) infection of nonproliferating human monocytes  

PubMed Central

Human immunodeficiency virus type 1 (HIV-1) infection of T lymphocytes requires cellular proliferation and DNA synthesis. Human monocytes were shown to have low DNA synthesis rates, yet the monocytotropic BaL isolate of HIV-1 was able to infect these cells efficiently. Monocytes that were irradiated to assure no DNA synthesis could also be readily infected with HIV-1BaL. Such infections were associated with the integration of HIV-1BaL DNA into the high molecular weight, chromosomal DNA of monocytes. Thus, normal, nonproliferating monocytes differ from T lymphocytes in that a productive HIV-1 infection can occur independently of cellular DNA synthesis. These results suggest that normal nonproliferating mononuclear phagocytes, which are relatively resistant to the destructive effects of this virus, may serve as persistent and productive reservoirs for HIV-1 in vivo.



[In vitro study of the primary antibody response of circulating lymphocytes in patients with chronic inflammatory rheumatism].  


We have studied the in vitro antibody response to a hapten of peripheral blood lymphocytes from 26 patients with rheumatoid arthritis and 7 ankylosing spondylitis. These patients had never received immunosuppressor drugs before or corticosteroids during the month before the test. They had failed to receive aspirin or non-steroid anti-inflammatory drugs for 72 hours before blood sampling. The control groups included respectively 38 healthy subjects and 24 patients hospitalized for non inflammatory disease. The antibody response of ankylosing spondylitis patients is comparable to that of controls ; on the opposite the response of patients with rhumatoid arthritis is significantly depressed in comparison with the three other groups. The weak response of lymphocytes in arthritis is not due to increased cell death in culture or to modified kinetics of the antibody response or to the appearance of a IgG secondary type response or a in vivo pre-activation. The lymphocytes of arthritis patients do not inhibit the response of normal lymphocytes when they are co-cultured. The observed response is identical to that obtained when control patient lymphocytes are co-cultured with normal lymphocytes. The function of suppressor T cells induced by Con A seems normal in spondylitis and arthritis. PMID:161063

Segond, P; Delfraissy, J F; Galanaud, P; Dormont, J; Massias, P



Lack of autologous mixed lymphocyte reaction in patients with chronic lymphocytic leukemia: evidence for autoreactive T-cell dysfunction not correlated with phenotype, karyotype, or clinical status  

SciTech Connect

In the present study, there was a complete lack of autologous MLR between responding T cells or T subsets and unirradiated or irradiated leukemic B cells or monocytes in all 20 patients with CLL, regardless of disease status, stage, phenotype, or karyotype of the disease. The stimulating capacity of unirradiated CLL B cells and CLL monocytes or irradiated CLL B cells was significantly depressed as compared to that of respective normal B cells and monocytes in allogeneic MLR. The responding capacity of CLL T cells was also variably lower than that of normal T cells against unirradiated or irradiated normal allogeneic B cells and monocytes. The depressed allogeneic MLR between CLL B cells or CLL monocytes and normal T cells described in the present study could be explained on the basis of a defect in the stimulating antigens of leukemic B cells or monocytes. The decreased allogeneic MLR of CLL T cells might simply be explained by a defect in the responsiveness of T lymphocytes from patients with CLL. However, these speculations do not adequately explain the complete lack of autologous MLR in these patients. When irradiated CLL B cells or irradiated CLL T cells were cocultured with normal T cells and irradiated normal B cells, it was found that there was no suppressor cell activity of CLL B cells or CLL T cells on normal autologous MLR. Our data suggest that the absence or dysfunction of autoreactive T cells within the Tnon-gamma subset account for the lack of autologous MLR in patients with CLL. The possible significance of the autologous MLR, its relationship to in vivo immunoregulatory mechanisms, and the possible role of breakdown of autoimmunoregulation in the oncogenic process of certain lymphoproliferative and autoimmune diseases in man are discussed.

Han, T.; Bloom, M.L.; Dadey, B.; Bennett, G.; Minowada, J.; Sandberg, A.A.; Ozer, H.



Secretion of phospholipase A 2 induced by interactions of human platelets with monocytes  

Microsoft Academic Search

Summary Phospholipase A2 (PLA2) activity was found and measured in the cell-free supernatants of human mononuclear cells (monocytes and lymphocytes) cultured with platelets for 48 h at 37° C. The relative molecular mass of purified, calcium-dependent PLA2 was 14 kD. The amount of PLA2 in the supernatants correlated positively with the number of monocytes and platelets in the cultures. Electron

S. Sipka; T. Farkas; P. Gergely; L. Bali; J. Laczko; S. Szabados; I. Csipo; M. Koltai; G. Szegedi



Monocyte chemoattractant protein-1 serum levels in ovarian cancer patients  

Microsoft Academic Search

The chemokine monocyte chemoattractant protein (MCP)-1 is an important mediator of monocyte infiltration in various solid tumours of epithelial origin. The aim of the present study was to evaluate the role of MCP-1 in the natural history of ovarian cancer and to determine its value as differentiation marker and prognostic marker regarding disease free and overall survival. This retrospective study

L Hefler; C Tempfer; G Heinze; K Mayerhofer; G Breitenecker; S Leodolter; A Reinthaller; C Kainz



Phase II study of theophylline in chronic lymphocytic leukemia: a study of the Eastern Cooperative Oncology Group (E4998)  

Microsoft Academic Search

The Eastern Cooperative Oncology Group (ECOG) performed a phase 2 study in B-cell chronic lymphocytic leukemia (CLL) of oral theophylline, a methylxanthine that inhibits cyclic nucleotide phosphodiesterases, thereby inducing the intracellular accumulation of cyclic adenosine monophosphate (cAMP). In 25 patients with Rai stages 0–I, theophylline, 200 mg given orally every 12 h was well tolerated. There was one complete response

P H Wiernik; E Paietta; O Goloubeva; S J Lee; D Makower; J M Bennett; J L Wade; C Ghosh; L S Kaminer; J Pizzolo; M S Tallman



Protective effect of grape seed extracts on human lymphocytes: a preliminary study.  


Grape seed extracts (GSEs) possess a broad spectrum of antioxidative properties that protects various cells from free radicals and oxidative stress. In this study, the genoprotective effect of GSE on human lymphocytic DNA was studied using standard and lysed cell comet assays. Lymphocytes from 5 healthy subjects were pretreated with GSE in different concentrations. The standard and lysed cell comet assays were performed on treated, untreated, challenged, and unchallenged cells in parallel. Cells were then subjected to an oxidant challenge induced with 5-min exposures to hydrogen peroxide. In the standard comet assay, GSE significantly diminished hydrogen-peroxide-induced DNA damage in a dose-dependent manner. In the lysed cell assay, however, the antioxidant effect was diminished at a higher GSE concentration. Data indicate that the cell membrane might play a role in limiting cellular access to antioxidants, which directly affects the genoprotective or potential pro-oxidant effect of antioxidants on human DNA. Using both standard and lysed cell comet assays in parallel could be a useful way to elucidate the mechanism of protection or damage by antioxidants. PMID:23537018

Szeto, Yim Tong; Lee, Kit Yee; Kalle, Wouter; Pak, Sok Cheon



Phase II study of theophylline in chronic lymphocytic leukemia: a study of the Eastern Cooperative Oncology Group (E4998).  


The Eastern Cooperative Oncology Group (ECOG) performed a phase 2 study in B-cell chronic lymphocytic leukemia (CLL) of oral theophylline, a methylxanthine that inhibits cyclic nucleotide phosphodiesterases, thereby inducing the intracellular accumulation of cyclic adenosine monophosphate (cAMP). In 25 patients with Rai stages 0-I, theophylline, 200 mg given orally every 12 h was well tolerated. There was one complete response after 22.5 months of treatment, which continues at 27+ months, and 18 other patients had stable disease. In vitro exposure of patients' lymphocytes to aminophylline (75-250 microg/ml), the soluble form of theophylline, resulted in dose- and time-dependent induction of apoptosis in 9/20 patients studied. Apoptosis was documented flow-cytometrically by monitoring the expression of bcl-2 and bax, forward light scatter, fluorescence intensity of binding of CD45 antibody, and the binding of annexin. Patients whose leukemic lymphocytes were susceptible to apoptosis induction by aminophylline in vitro experienced a significantly longer progression-free survival than patients whose cells were resistant to the drug in culture (P=0.025). This suggests that in a CLL population treated with theophylline, induction of an apoptotic response to the drug in vitro is prognostic for absence of clinical progression. PMID:15356646

Wiernik, P H; Paietta, E; Goloubeva, O; Lee, S J; Makower, D; Bennett, J M; Wade, J L; Ghosh, C; Kaminer, L S; Pizzolo, J; Tallman, M S



Collagenous and lymphocytic colitis.  


Collagenous and lymphocytic colitis have been recognized as chronic intestinal inflammatory disorders causing watery diarrhea, which have been recognized in the past three to two decades, respectively. Collagenous colitis is primarily a disorder of middle-aged women and is characterized on biopsy by increased subepithelial collagen as well as increased inflammatory cells in the lamina propria and increased intraepithelial lymphocytes. Key to the correct diagnosis in this condition is recognizing that there are two words in this diagnostic entity, and colitis is, by definition, present. Focusing solely on the collagen band can result in both over- and underdiagnosis. Newer therapeutic options are available in this condition, and patients are now frequently being treated either with budesonide or with high dose bismuth preparations. Whereas collagenous colitis is a tightly coherent clinical pathologic entity, lymphocytic colitis has a more varied clinical picture. Lymphocytic colitis is also seen in middle-aged patients but has a more equal female-to-male ratio. Lymphocytic colitis is defined by increased intraepithelial lymphocytes, with the median being 30 lymphocytes per 100 epithelial cells. There are also an increase in inflammatory cells in the lamina propria, but the increase may be milder than in collagenous colitis and there are usually minimal eosinophils. Although numerous studies have described lymphocytic colitis causing a chronic diarrhea, more recent studies suggest that patients may have a single attack in approximately 60% of cases. Although most cases of lymphocytic colitis are idiopathic, there is a clear association with multiple drugs, celiac disease, and there may be an infectious trigger. Approximately 10% of lymphocytic colitis patients have a positive family history of some type of inflammatory intestinal disease, including ulcerative colitis, Crohn's disease, collagenous colitis, and celiac disease. Therapy in lymphocytic colitis is less well studied, but the same medications are used with success, including budesonide and high dose bismuth. PMID:16939057

Lazenby, Audrey J



BCR signaling in chronic lymphocytic leukemia and related inhibitors currently in clinical studies.  


Normal B lymphocytes receive signals from B-cell antigen receptor (BCR) that are triggered by binding of the BCR to an external antigen. Tonic signaling through the BCR provides growth and signals to chronic lymphocytic leukemia (CLL) cells, and plays an important role in the pathogenesis and progression of the disease. Antigen engagement of BCR is followed by intracellular recruitment and activation of BCR-associated kinases including spleen tyrosine kinase (Syk), Bruton's tyrosine kinase (Btk) and phosphatidylinositol 3-kinases (PI3K). Inhibition of signaling pathways downstream of the BCR induces disruption of chemokine-mediated CLL cell migration and cell killing. BCR signal transduction inhibitors represent a promising new strategy for targeted CLL treatment. A number of therapeutic agents have recently been developed with significant activity in CLL. The compounds that are currently investigated in patients with CLL include ibrutinib -inhibitor of Btk, fostamatinib-inhibitor of Syk and idelalisib (GS-1101) -a specific isoform of the PI3K (PI3K) inhibitor. The clinical activity of ibrutinib, GS-1101 and fostamatinib in patients with CLL is associated with marked lymphocytosis due to release of tumor cells from the lymph nodes into the peripheral blood. Further studies are ongoing with single agents and their combinations with other targeted and conventional therapies. This article will review the preclinical rationale of BCR signaling inhibitors in the treatment of CLL, and the clinical evidence supporting the use of these agents in CLL patients. PMID:23617253

Robak, Tadeusz; Robak, Pawel



Polarity and sensitivity of T lymphocyte studied by an optical trap  

NASA Astrophysics Data System (ADS)

Lymphocytes are the central players in the human adaptive immune response. In the body, individual T helper lymphocytes need to be activated first by physical contact with antigen- presenting cells (APC). T-cell contact with APCs initiated an activation cascade, which includes an increase in T-cell intracellular calcium, leading to T-cell proliferation, differentiation and lymphokine production. Calcium imaging are combined with optical manipulation to investigate the physical properties of T-cell activation. We study cell-cell contact requirements for T-cell activation using optical tweezers to control the orientation of T-cell/APC pairs and fluorescence microscopy to measure the subsequent T-cell intracellular calcium level [(Ca2+)i] response. APCs or beads coated with antibodies to the T-cell receptor (TCR) are trapped with a near-infrared titanium-sapphire laser and placed at different locations along the T-cell, which has a polarized appearance defined by the shape and direction of crawling. T cells which are presented with antigen at the leading edge have a higher probability of responding and a shorter latency of response than those contacting APCs or beads with their trailing end. Alterations in antibody density and bead size are used to determine the spatial requirements for T cell activation and the minimum number of receptors which must be engaged in order to transmit a positive signal.

Wei, Xunbin; Krasieva, Tatiana B.; Cahalan, Michael D.; Tromberg, Bruce J.



Jumping translocation in a phenotypically normal male: A study of mosaicism in spermatozoa, lymphocytes, and fibroblasts.  


Both Robertsonian translocations, rob(13;13) and rob(13;15), (in the present case defined as dic(13;15)), are rare chromosomal rearrangements and there is scarce information regarding their behavior during meiosis. In this report we describe a man with mosaicism for two cell lines, each cell line containing a different de novo Robertsonian translocation with the common breakpoint in the centromeric region on chromosome 13. The karyotype was finally defined as: 45,XY,rob(13;13)(q10;q10)[29]/45,XY,dic(13;15)(p11.2;p12)[22], a phenomenon referred to as jumping translocation. The relative occurrence of the two clones in lymphocytes and fibroblasts as well as the meiotic segregation in spermatozoa and the mechanism of formation were studied using karyotype analysis, fluorescence in situ hybridization (FISH), and quantitative fluorescence-PCR. Karyotype analysis of cultured lymphocytes revealed 57% rob(13;13) cells and 43% dic(13;15) cells and for cultured skin fibroblasts the figures were almost identical (56% and 44%, respectively). FISH analysis showed 55% balanced nuclei for unselected spermatozoa and after swim-up selection the number of balanced spermatozoa decreased to 41%. In addition, 16% of the unselected spermatozoa and 27% of the spermatozoa after swim-up selection carried an additional chromosome 13, indicating a high risk for a trisomy 13 offspring. Swim-up selection did not increase the number of balanced spermatozoa. PMID:19610103

Iwarsson, Erik; Sahlén, Sigrid; Nordgren, Ann



Rho proteins, PI 3-kinases, and monocyte\\/macrophage motility  

Microsoft Academic Search

Rho proteins and phosphatidylinositide 3-kinases (PI 3-kinases) have been widely implicated in regulating cell motility both in cultured cells and in animal models. Monocytes are recruited from the bloodstream in response to inflammatory signals, and migrate across the endothelial barrier into the tissues, where they differentiate into macrophages and phagocytose bacteria and cells. Studies of monocytes and macrophages have revealed

Anne J. Ridley



Tumor cell-induced deactivation of human monocytes  

Microsoft Academic Search

Although blood monocytes exhibit sig- nificant cytotoxic activity against tumor cells, the function of tumor infiltrating macrophages (TIM) is depressed in cancer patients. This study addresses the question of how the antitumor response of hu- man monocytes, assessed by production of cyto- kines (tumor necrosis factor , TNF; IL-10; IL- 12p40) and cytotoxicity, is altered by exposure to cancer cells.

Bozenna Mytar; Maria Wooszyn; Monika Baj-Krzyworzeka; Maciej Siedlar; Irena Ruggiero; Marek Zembala



Immunomodulating activity of seaweed extract on human lymphocytes in vitro.  


Effect of eight kinds of seaweed extract (SWE) on human lymphocytes was studied in vitro. The extracts of Hizikiafusiformis and Meristotheca papulosa (green) markedly stimulated human lymphocytes to proliferate, whereas Eucheuma muricatum and Meristotheca papulosa (red) weakly stimulated proliferation. The responder cells are T cells, because T cells purified by sheep red blood cell (SRBC) rosette-formation were significantly stimulated with SWE, but B cells were not. These extracts enhanced the induction of cytotoxic T lymphocyte (CTL) activity, but failed to enhance natural killer (NK) cell activity. These extracts had a stimulatory effect on immunoglobulin (Ig) production by B cells and tumor necrosis factor (TNF) production by monocytes. The activity of Hizikia fusiformis associated with polysaccharides which were extracted with ethanol and purified by ion-exchange and gel filtration chromatography, whose molecular weight was about 100 kDa. These results suggest that SWE has an immunomodulating activity on human lymphocytes and this ability might be useful for clinical application to treat several diseases such as tumors. PMID:10411282

Shan, B E; Yoshida, Y; Kuroda, E; Yamashita, U



Lymphocyte apoptosis in children with central nervous system tuberculosis: a case control study  

PubMed Central

Background Studies of the apoptosis mechanisms involved in the pathogenesis of tuberculosis have suggested that Mycobacterium tuberculosis can actively interfere with the apoptosis of infected cells. In vivo studies have been performed in adult populations but have not focused on this process in children. In the present study, we analyzed spontaneous T lymphocyte (PBT) apoptosis in the peripheral blood of children with central nervous system tuberculosis (CNS TB), before and after chemotherapy, and compared the results with healthy controls. Methods A case-control study was conducted from January 2002 to June 2009. It included 18 children with CNS TB and 17 healthy controls. Spontaneous apoptosis of PBTs, including CD4+, CD8+ and CD8+/CD28+ T cells, was evaluated after 24 and 72 h of culture in complete medium, using the Annexin V detection test. Analysis was conducted before and after chemotherapy, and expression of the apoptotic markers CD95 (Fas) and Fas ligand (FasL) was evaluated. Results Higher percentages of apoptotic T cells and CD4 lymphocytes were isolated from children with acute phase CNS TB than from children in the control group (p < 0.05). This difference significantly decreased after 60 days of specific treatment. In children with CNS TB, high levels of Fas ligand expression were detected in lymphocyte populations, associated with a high percentage of Fas positive cells, before and after treatment. In contrast to the CD4+ apoptosis profile, we did not find any significant difference in total CD8+ cell apoptosis between children with acute phase disease and the control group. However, the percentage of apoptotic CD8+/CD28+ T cells was significantly higher in the children with acute phase disease than in the healthy controls. Conclusions Our findings indicate that CNS TB in pediatric patients increases the sensitivity of CD4 and CD8+/CD28+ T cells to apoptosis, suggesting a hypoergic status of this infection. This could play a key role in the immunopathogenesis of this complicated form of TB. Interestingly, specific chemotherapy is able to normalize both apoptosis sensitivity and T-cell activation.



Genome-Wide Association Study Identifies Single Nucleotide Polymorphism in DYRK1A Associated with Replication of HIV-1 in Monocyte-Derived Macrophages  

PubMed Central

Background HIV-1 infected macrophages play an important role in rendering resting T cells permissive for infection, in spreading HIV-1 to T cells, and in the pathogenesis of AIDS dementia. During highly active anti-retroviral treatment (HAART), macrophages keep producing virus because tissue penetration of antiretrovirals is suboptimal and the efficacy of some is reduced. Thus, to cure HIV-1 infection with antiretrovirals we will also need to efficiently inhibit viral replication in macrophages. The majority of the current drugs block the action of viral enzymes, whereas there is an abundance of yet unidentified host factors that could be targeted. We here present results from a genome-wide association study identifying novel genetic polymorphisms that affect in vitro HIV-1 replication in macrophages. Methodology/Principal Findings Monocyte-derived macrophages from 393 blood donors were infected with HIV-1 and viral replication was determined using Gag p24 antigen levels. Genomic DNA from individuals with macrophages that had relatively low (n?=?96) or high (n?=?96) p24 production was used for SNP genotyping with the Illumina 610 Quad beadchip. A total of 494,656 SNPs that passed quality control were tested for association with HIV-1 replication in macrophages, using linear regression. We found a strong association between in vitro HIV-1 replication in monocyte-derived macrophages and SNP rs12483205 in DYRK1A (p?=?2.16×10?5). While the association was not genome-wide significant (p<1×10?7), we could replicate this association using monocyte-derived macrophages from an independent group of 31 individuals (p?=?0.0034). Combined analysis of the initial and replication cohort increased the strength of the association (p?=?4.84×10?6). In addition, we found this SNP to be associated with HIV-1 disease progression in vivo in two independent cohort studies (p?=?0.035 and p?=?0.0048). Conclusions/Significance These findings suggest that the kinase DYRK1A is involved in the replication of HIV-1, in vitro in macrophages as well as in vivo.

Bol, Sebastiaan M.; Moerland, Perry D.; Limou, Sophie; van Remmerden, Yvonne; Coulonges, Cedric; van Manen, Danielle; Herbeck, Joshua T.; Fellay, Jacques; Sieberer, Margit; Sietzema, Jantine G.; van 't Slot, Ruben; Martinson, Jeremy; Zagury, Jean-Francois; Schuitemaker, Hanneke; van 't Wout, Angelique B.



Monocyte attachment to laminin in diabetes mellitus: The role of ATP.  


Monocyte-extracellular matrix interactions have been implicated in atherosclerosis pathophysiology. Laminin, the main basement membrane protein contains cell binding domains that can be cryptic, presented only after protein modification. In the present study we evaluated monocyte attachment to laminin-1 in the presence of ATP. Monocytes were derived from either healthy volunteers or patients with diabetes mellitus type II. For the estimation of monocyte attachment to laminin the myeloperoxidase assay was used. Monocytes derived from diabetic patients, showed an increased ability to attach to laminin (p = 0.0055). The presence of ATP increased the attachment of control monocytes to laminin (p = 0.0022). On the contrary, the presence of ATP did not affect the attachment of monocytes derived from diabetic patients to laminin. Our results indicate a modified interaction between monocytes and laminin-1 in diabetes mellitus. PMID:19287210

Kostidou, Elena; Trachana, Varvara; Topouridou, Konstantina; Paletas, Konstantinos; Tsapas, Apostolos; Kaloyianni, Martha; Koliakos, George



Chlamydia pneumoniae Infection in Human Monocytes  

Microsoft Academic Search

Chlamydia pneumoniae infection has been associated with cardiovascular diseases in seroepidemiological studies and by demonstration of the pathogen in atherosclerotic lesions. It has the capacity to infect several cell types, including monocyte-derived macrophages, which play an essential role in the development of athero- sclerosis. However, the persistence of C. pneumoniae in mononuclear cells is poorly understood. To study the morphology



Some immunological studies in aplastic anaemia.  


Peripheral blood studies in 29 patients with aplastic anaemia showed decrease in the mean numbers of total lymphocytes, T and B lymphocytes and monocytes. Quantitation of serum immunoglobulins carried out in 22 patients showed normal results in most cases. Although the significance of these immunological changes in the development of aplastic anaemia is not established they may represent a factor in addition to neutropenia in the pathogenesis of infections in this disease. PMID:6600793

Falcão, R P; Voltarelli, J C; Bottura, C



Retinal Pigment Epithelium Cells Promote the Maturation of Monocytes to Macrophages in vitro  

Microsoft Academic Search

Proliferative vitreoretinopathy is characterized by excessive cell proliferation within the eye; retinal pigment epithelial (RPE) cells form the majority of proliferating cells and interact with infiltrating leukocytes including monocytes. The purpose of this study was to determine the effect of RPE cells on the maturation of monocytes to macrophages. The enriched monocyte fraction of peripheral blood mononuclear cells was either

Roman Osuský; Punam Malik; Stephen J. Ryan



Stimulation of Human Monocytes with the Gram-Positive Vaccine Vector Streptococcus gordonii  

Microsoft Academic Search

Streptococcus gordonii is a bacterial vaccine vector which has previously been shown to activate dendritic cells in vitro and to induce local and systemic immune responses in vivo. In the present study, human monocytes (THP-1 cell line and peripheral blood monocytes) were characterized following interaction with S. gordonii. Treatment of human monocytes with S. gordonii but not latex beads induced

Annalisa Ciabattini; Anna Maria Cuppone; Rita Pulimeno; Francesco Iannelli; Gianni Pozzi; Donata Medaglini



Study of HLA class I restriction and the directed antigens of cytotoxic T lymphocytes at the tumor sites of ovarian cancer  

Microsoft Academic Search

The molecular basis of T-cell-mediated recognition of ovarian cancer cells remains to be fully addressed. In this study we\\u000a investigated HLA class I restriction and directed antigens of cytotoxic T lymphocytes (CTL) at the sites of ovarian cancer.\\u000a Three HLA-class-I-restricted CTL lines were established from the tumor sites of ovarian cancer by culturing tumor-infiltrating\\u000a lymphocytes or tumor-associated ascitic lymphocytes with

Akira Yamada; Koichiro Kawano; Nanae Harashima; Fumihiko Niiya; Kouji Nagai; Terutada Kobayashi; Takashi Mine; Kimio Ushijima; Takashi Nishida; Kyogo Itoh



Apoptosis induction of K562 cells by lymphocytes: an AFM study.  


Antitumor immunotherapies, as a prospective approach for local cancer treatment, are attracting increasing interests. To detect the reacting course of immune and tumor cells, we have observed the process of K562 cells (a human erythroleukemic cell line) coculturing with peripheral lymphocytes, and the morphological and ultrastructural alterations of K562 cells and lymphocytes were investigated as well using atomic force microscopy (AFM). AFM morphological imaging revealed that after coculture the apoptosis-like structures such as blebbing, pores, and apoptotic bodies were observed on the K562 cells. Also, the cell-surface roughness decreased significantly, which implied the changes in chemical composition of cell membranes. Moreover, the lymphocytes were damaged to some extent induced by the coculture. The data demonstrated that K562 cells could be attacked and induced apoptosis by lymphocytes, and they would make damages to lymphocytes to escape the surveillance of immune system. PMID:23417662

Jin, Hua; Zhao, Hongxia; Liu, Li; Jiang, Jinhuan; Wang, Xiaoping; Ma, Shuyuan; Cai, Jiye


Alterations in Monocyte CD16 in Association with Diabetes Complications  

PubMed Central

Monocytes express many cell surface markers indicative of their inflammatory and activation status. Whether these markers are affected by diabetes and its complications is not known and was investigated in this study. Blood was obtained from 22 nondiabetic and 43 diabetic subjects with a duration of diabetes >10 years, including 25 without and 18 with clinically significant complications. The number of CD45+CD14+ monocytes and the percentage expressing the proinflammatory marker CD16 were determined by flow cytometry. Other markers of monocyte activation and expression of chemokine receptors were also examined. The relationship between monocyte CD16 and clinical data, selected cytokines, and chemokines was also investigated. Diabetes had no effect on total white cell number but increased monocyte number. Diabetes also significantly decreased the number of CD16+ monocytes but only in those with diabetic complications. Other markers of monocyte activation status and chemokine receptors were not affected by diabetes or complications status. Diabetes induced plasma proinflammatory cytokines and they were lower in diabetic subjects with complications compared to those without complications. These results suggest that the circulating monocyte phenotype is altered by diabetic complications status. These changes may be causally related to and could potentially be used to predict susceptibility to diabetic complications.

Min, Danqing; Brooks, Belinda; Wong, Jencia; Salomon, Robert; Bao, Wensheng; Harrisberg, Brian; Twigg, Stephen M.; Yue, Dennis K.; McLennan, Susan V.



Secretion of monocyte chemotactic activity by alveolar macrophages.  

PubMed Central

The purpose of this study was to determine if alveolar macrophages (AMs) are a source of monocyte chemoattractants and the role bleomycin interaction with AMs may play in the recruitment of monocytes to the lung in a rodent model of bleomycin-induced pulmonary fibrosis. AMs isolated from rats with bleomycin-induced fibrosis secreted significantly greater amounts of monocyte chemoattractants than those isolated from normal rats. When AMs from normal rats were stimulated with bleomycin in vitro, monocyte chemotactic activity was secreted into the medium. Chemotactic activity secretion by AM stimulated with 0.01 to 0.1 micrograms/ml bleomycin was significantly higher than that of cells incubated in medium alone. This activity was truly chemotactic for monocytes, but caused only minimal migration of normal AMs. Bleomycin itself at concentrations of 1 pg/ml to 10 micrograms/ml had no monocyte chemoattractant activity. Characterization of the chemotactic activity in conditioned media (CM) from bleomycin-stimulated AM demonstrated that the major portion of the activity bound to gelatin, was heterogeneous, with estimated molecular weights of 20 to 60 kd, and was inactivated by specific antifibronectin antibody. These findings suggest that fibronectin fragments are primarily responsible for the monocyte chemotactic activity secreted by AMs. Through increased secretion of such chemotactic substances, AMs could play a key role in the recruitment of peripheral blood monocytes into the lung in inflammatory lung disease and fibrosis.

Denholm, E. M.; Wolber, F. M.; Phan, S. H.



Blood monocytes: distinct subsets, how they relate to dendritic cells, and their possible roles in the regulation of T-cell responses  

Microsoft Academic Search

Monocytes can have important effects on the polarization and expansion of lymphocytes and may contribute to shaping primary and memory T-cell responses in humans and mice. However, their precise contribution in terms of cellular subsets and the molecular mechanisms involved remains to be determined. Mouse monocytes originate from a bone marrow progenitor, the macrophage and DC precursor (MDP), which also

Frederic Geissmann; Cedric Auffray; Roger Palframan; Christiane Wirrig; Alice Ciocca; Laura Campisi; Emilie Narni-Mancinelli; Gregoire Lauvau



Monocytes and their pathophysiological role in Crohn's disease.  


Our immune system shows a stringent dichotomy, on the one hand displaying tolerance towards commensal bacteria, but on the other hand vigorously combating pathogens. Under normal conditions the balance between flora tolerance and active immunity is maintained via a plethora of dynamic feedback mechanisms. If, however, the balancing act goes faulty, an inappropriate immune reaction towards an otherwise harmless intestinal flora causes disease, Crohn's disease for example. Recent developments in the immunology and genetics of mucosal diseases suggest that monocytes and their derivative cells play an important role in the pathophysiology of Crohn's disease. In our review, we summarize the recent studies to discuss the dual function of monocytes - on the one hand the impaired monocyte function initiating Crohn's disease, and on the other hand the overactivation of monocytes and adaptive immunity maintaining the disease. With a view to developing new therapies, both aspects of monocyte functions need to be taken into account. PMID:18791847

Zhou, L; Braat, H; Faber, K N; Dijkstra, G; Peppelenbosch, M P



Childhood nephrotic syndrome in relapse is associated with down-regulation of monocyte CD14 expression and lipopolysaccharide-induced tumour necrosis factor-? production  

PubMed Central

Interleukin-13 (IL-13) is a known modulator of monocyte function, down-regulating monocyte surface markers such as CD14 and proinflammatory cytokines. We have shown previously that lymphocyte IL-13 gene expression was up-regulated during relapses in children with steroid-responsive nephrotic syndrome (SRNS). In this study, we examined the monocyte mRNA expression and lipopolysaccharide (LPS)-stimulated intracellular production of tumour necrosis factor-? (TNF-?) and IL-8 in children with SRNS during relapse and remission. Additionally, we investigated CD14 mRNA levels, CD14 surface expression and its soluble component (sCD14) in serum. Our results showed that the percentages of TNF-? positive monocytes following LPS stimulation were significantly lower in nephrotic children in relapse (64·4 ± 13·7%) compared to remission (81·6 ± 9·0%, P < 0·005). This was associated with down-regulation of CD14 mRNA, as well as both membrane and sCD14 in patients with nephrotic relapse (82·9 ± 10·1% and 1·23 ± 0·30 µg/ml, respectively) compared to remission (93·9 ± 3·2% and 1·77 ± 0·82 µg/ml, respectively) (P < 0·003). Although we demonstrated a decrease in LPS-stimulated intracellular production of TNF-? in monocytes from patients with nephrotic relapse, we were unable to show a concomitant decrease in mRNA expression during relapses. This could be explained by down-regulation of gene expression at the translational rather than transcriptional level. In conclusion, it is conceivable that up-regulation of T-cell IL-13 production in children with active nephrotic relapse was associated with suppression of monocyte CD14 expression, down-regulating pro-inflammatory cytokine production, and could account for the increased susceptibility to bacterial sepsis seen in nephrotic children in active relapse.




Induction of reactive oxygen intermediates in human monocytes by tumour cells and their role in spontaneous monocyte cytotoxicity  

PubMed Central

The present study examined the ability of human monocytes to produce reactive oxygen intermediates after a contact with tumour cells. Monocytes generated oxygen radicals, as measured by luminol-enhanced chemiluminescence and superoxide anion production, after stimulation with the tumour, but not with untransformed, cells. The use of specific oxygen radical scavengers and inhibitors, superoxide dismutase, catalase, dimethyl sulphoxide and deferoxamine as well as the myeloperoxidase inhibitor 4-aminobenzoic acid hydrazide, indicated that chemiluminescence was dependent on the production of superoxide anion and hydroxyl radical and the presence of myeloperoxidase. The tumour cell-induced chemiluminescent response of monocytes showed different kinetics from that seen after activation of monocytes with phorbol ester. These results indicate that human monocytes can be directly stimulated by tumour cells for reactive oxygen intermediate production. Spontaneous monocyte-mediated cytotoxicity towards cancer cells was inhibited by superoxide dismutase, catalase, deferoxamine and hydrazide, implicating the role of superoxide anion, hydrogen peroxide, hydroxyl radical and hypohalite. We wish to suggest that so-called ‘spontaneous’ tumoricidal capacity of freshly isolated human monocytes may in fact be an inducible event associated with generation of reactive oxygen intermediates and perhaps other toxic mediators, resulting from a contact of monocytes with tumour cells. © 1999 Cancer Research Campaign

Mytar, B; Siedlar, M; Woloszyn, M; Ruggiero, I; Pryjma, J; Zembala, M



Human Bladder Uroepithelial Cells Synergize with Monocytes to Promote IL-10 Synthesis and Other Cytokine Responses to Uropathogenic Escherichia coli  

PubMed Central

Urinary tract infections are a major source of morbidity for women and the elderly, with Uropathogenic Escherichia coli (UPEC) being the most prevalent causative pathogen. Studies in recent years have defined a key anti-inflammatory role for Interleukin-10 (IL-10) in urinary tract infection mediated by UPEC and other uropathogens. We investigated the nature of the IL-10-producing interactions between UPEC and host cells by utilising a novel co-culture model that incorporated lymphocytes, mononuclear and uroepithelial cells in histotypic proportions. This co-culture model demonstrated synergistic IL-10 production effects between monocytes and uroepithelial cells following infection with UPEC. Membrane inserts were used to separate the monocyte and uroepithelial cell types during infection and revealed two synergistic IL-10 production effects based on contact-dependent and soluble interactions. Analysis of a comprehensive set of immunologically relevant biomarkers in monocyte-uroepithelial cell co-cultures highlighted that multiple cytokine, chemokine and signalling factors were also produced in a synergistic or antagonistic fashion. These results demonstrate that IL-10 responses to UPEC occur via multiple interactions between several cells types, implying a complex role for infection-related IL-10 during UTI. Development and application of the co-culture model described in this study is thus useful to define the degree of contact dependency of biomarker production to UPEC, and highlights the relevance of histotypic co-cultures in studying complex host-pathogen interactions.

Duell, Benjamin L.; Carey, Alison J.; Dando, Samantha J.; Schembri, Mark A.; Ulett, Glen C.



Human Bladder Uroepithelial Cells Synergize with Monocytes to Promote IL-10 Synthesis and Other Cytokine Responses to Uropathogenic Escherichia coli.  


Urinary tract infections are a major source of morbidity for women and the elderly, with Uropathogenic Escherichia coli (UPEC) being the most prevalent causative pathogen. Studies in recent years have defined a key anti-inflammatory role for Interleukin-10 (IL-10) in urinary tract infection mediated by UPEC and other uropathogens. We investigated the nature of the IL-10-producing interactions between UPEC and host cells by utilising a novel co-culture model that incorporated lymphocytes, mononuclear and uroepithelial cells in histotypic proportions. This co-culture model demonstrated synergistic IL-10 production effects between monocytes and uroepithelial cells following infection with UPEC. Membrane inserts were used to separate the monocyte and uroepithelial cell types during infection and revealed two synergistic IL-10 production effects based on contact-dependent and soluble interactions. Analysis of a comprehensive set of immunologically relevant biomarkers in monocyte-uroepithelial cell co-cultures highlighted that multiple cytokine, chemokine and signalling factors were also produced in a synergistic or antagonistic fashion. These results demonstrate that IL-10 responses to UPEC occur via multiple interactions between several cells types, implying a complex role for infection-related IL-10 during UTI. Development and application of the co-culture model described in this study is thus useful to define the degree of contact dependency of biomarker production to UPEC, and highlights the relevance of histotypic co-cultures in studying complex host-pathogen interactions. PMID:24155979

Duell, Benjamin L; Carey, Alison J; Dando, Samantha J; Schembri, Mark A; Ulett, Glen C



Stimulatory and inhibitory influences of human immunodeficiency virus on normal B lymphocytes.  

PubMed Central

B-lymphocyte dysfunction is a characteristic feature of the acquired immunodeficiency syndrome (AIDS) and of the AIDS-related complex. The aim of the present study was to further examine the influences exercised by the human immunodeficiency virus (HIV; formerly called human T-lymphotropic virus type III or lymphadenopathy-associated virus, HTLV-III/LAV) on normal human B lymphocytes. An unfractionated protein preparation, made from HIV purified by density gradient centrifugation, was previously shown to induce differentiation of normal human B lymphocytes into immunoglobulin-secreting cells. In the present analyses, this B-lymphocyte response peaked on day 6 or 7 after culture initiation and was found to be independent of the requirement for monocytes but to require T cells. Responses could also be elicited in cultures of purified B cells by the addition of T cells that had been exposed to HIV antigen. Inhibitors of protein synthesis (puromycin and cycloheximide) abrogated the responses. In contrast to its stimulatory effects, the same virus preparation was previously shown to inhibit polyclonal responses that are normally elicited in peripheral blood lymphocyte cultures by a T-dependent stimulus (pokeweed mitogen) and T-independent stimulus (Epstein-Barr virus). The present studies suggest that the inhibitory effects of the HIV antigen studied herein are targeted primarily at the B lymphocytes. The role of T lymphocytes in the HIV antigen-mediated inhibitory effects, although demonstrated, could not be conclusively established as an essential pathway. These findings elucidate mechanisms by which components of HIV exert stimulatory as well as inhibitory effects on human B lymphocytes and thereby lead to the dysfunction of these cells in HIV infection.

Pahwa, S; Pahwa, R; Good, R A; Gallo, R C; Saxinger, C



Comparative studies of mitogen- and antigen-induced lymphocyte proliferation in four captive rhinoceros species.  


Cellular immune function in four rhinoceros species was evaluated by way of in vitro lymphocyte proliferation responses to mitogenic and antigenic stimuli to establish normative data on white blood cell activity for each species and to identify species-specific differences that might help explain the predisposition of black rhinoceroses (Diceros bicornis) to disease. A cross section of the U.S. rhinoceros population encompassing all four captive species was sampled, including the Sumatran rhinoceros (Dicerorhinus sumatrensis) (n = 3); Indian rhinoceros (Rhinoceros unicornis) (n = 4); African black rhinoceros (n = 16); and African white rhinoceros (Ceratotherium simum) (n = 10). Of the four species evaluated, African black rhinoceroses exhibited the weakest (P < 0.05) lymphocyte proliferative responses to the mitogens: pokeweed (0.1 microg/ml), phytohemagglutinin (0.3 microg/ml), and concanavalin A (5.0 microg/ml). Total cell density at the end of culture was only 70% of that achieved with lymphocytes isolated from African white rhinoceroses, Indian rhinoceroses, and Sumatran rhinoceroses. However, lymphocyte response to bacterial endotoxin lipopolysaccharide was similar (P > 0.05) across species. Antigenic stimulation produced much weaker responses than mitogenic stimulation. No differences (P > 0.05) were observed among rhinoceros species in response to 1 and 10 microg/ml of Leptospira icterohemorrhagiae or Leptospira gryppotyphosa. Lymphocytes from African white rhinoceroses proliferated weakly in the presence of Aspergillus fumigatus filtrate, whereas lymphocytes from the southern black rhinoceros subspecies appeared slightly suppressed in the presence of increasing doses (0.1, 1, and 10 microg/ml) of Aspergillus filtrate. This comparative data set characterizing lymphocyte proliferation in the rhinoceros reveals several differences in immune cell responses among rhinoceros species and provides some evidence that lymphocytes of captive African black rhinoceroses are less vigorous than those of the other rhinoceros species. PMID:15732584

Vance, Carrie K; Kennedy-Stoskopf, Suzanne; Obringer, Amy R; Roth, Terri L



Isolation of porcine monocyte population: a simple and efficient method.  


Monocytes are important mediators of inflammatory processes and are in the focus of immunological studies. While the preparation of human monocytes is widely established, little is published on the isolation of porcine monocytes for experimental studies. The aim of this study is to establish a cost efficient method of preparing and culturing porcine monocytes of considerable purity and reasonable yield. In our method, we combined and modified different protocols of human monocyte preparation. The blood of a single pig is harvested and treated with EDTA to prevent coagulation. Peripheral blood mononuclear cells are obtained by a density gradient centrifugation using a Bicoll gradient and monocytes are harvested by culturing on serum-treated culture flasks, rinsing and tapping. A high yield is obtained by constant cooling of flasks and tubes. The purity of the culture is evaluated by the expression of CD14, using flow cytometry. Using this method, we reached a purity of 92.6 % (+/- 3.06 %). With this procedure, we established a reliable method to prepare and cultivate porcine monocytes which can be performed cost effectively and does not require special equipment. PMID:23624769

Berg, Christin; Wilker, Sebastian; Roider, Johann; Klettner, Alexa



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


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

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



Bacillus anthracis Peptidoglycan Stimulates an Inflammatory Response in Monocytes through the p38 Mitogen-Activated Protein Kinase Pathway  

PubMed Central

We hypothesized that the peptidoglycan component of B. anthracis may play a critical role in morbidity and mortality associated with inhalation anthrax. To explore this issue, we purified the peptidoglycan component of the bacterial cell wall and studied the response of human peripheral blood cells. The purified B. anthracis peptidoglycan was free of non-covalently bound protein but contained a complex set of amino acids probably arising from the stem peptide. The peptidoglycan contained a polysaccharide that was removed by mild acid treatment, and the biological activity remained with the peptidoglycan and not the polysaccharide. The biological activity of the peptidoglycan was sensitive to lysozyme but not other hydrolytic enzymes, showing that the activity resides in the peptidoglycan component and not bacterial DNA, RNA or protein. B. anthracis peptidoglycan stimulated monocytes to produce primarily TNF?; neutrophils and lymphocytes did not respond. Peptidoglycan stimulated monocyte p38 mitogen-activated protein kinase and p38 activity was required for TNF? production by the cells. We conclude that peptidoglycan in B. anthracis is biologically active, that it stimulates a proinflammatory response in monocytes, and uses the p38 kinase signal transduction pathway to do so. Given the high bacterial burden in pulmonary anthrax, these findings suggest that the inflammatory events associated with peptidoglycan may play an important role in anthrax pathogenesis.

Langer, Marybeth; Malykhin, Alexander; Maeda, Kenichiro; Chakrabarty, Kaushik; Williamson, Kelly S.; Feasley, Christa L.; West, Christopher M.; Metcalf, Jordan P.; Coggeshall, K. Mark



Inability of newborns' or pregnant women's monocytes to suppress pokeweed mitogen-induced responses  

SciTech Connect

Although an excess of human adult blood adherent cells inhibits the pokeweed mitogen- (PWM) induced normal adult lymphocyte proliferation and B cell maturation into immunoglobulin-containing cells (ICC), adherent cells collected from newborn infants or pregnant women at time of delivery were unable to exert a similar suppressor activity. After activation by Concanavalin A (Con A), newborns' and pregnant women's adherent cells acquired a suppressor activity comparable to that of control adult adherent cells. The adherent suppressor cell was shown to be radioresistant (3000 rad), indicating its probable monocytic orgin. Both monocyte-suppressor activities (MSA) observed in adulthood (spontaneously) and in the neonatal period (after activation) were dependent on prostaglandin E/sub 2/ (PGE/sub 2/) secretion, because they were abolished by indomethacin or a specific anti-PGE/sub 2/ anti-serum. Expression of MSA appeared to be under a negative regulation exerted by naturally occurring T suppressor lymphocytes present in the blood of newborns or pregnant women, because incubation of adult monocytes or Con A-activated newborn monocytes with newborns' or pregnant women's T lymphocytes resulted in a dramatic decrease of their MSA. These results strongly suggest that the lack of MSA in the neonatal period and in late pregnancy is a consequence of activation of T suppressor lymphocytes.

Durandy, A.; Fischer, A.; Griscelli, C.



Phenotypic and functional heterogeneity of bovine blood monocytes.  


Murine and human peripheral blood monocytes are heterogeneous in size, granularity, nuclear morphology, phenotype and function. Whether and how bovine blood monocytes follow this pattern was analyzed in this study. Flow cytometrically, classical monocytes (cM) CD14?CD16?, intermediate monocytes (intM) CD14? CD16? and nonclassical monocytes (ncM) CD14? CD16? were identified, with cM being the predominant subset (89%). cM showed a significant lower expression of CD172a, intM expressed the highest level of MHC class II molecules, and ncM were low positive for CD163. Compared to cM and intM, ncM showed a significantly reduced phagocytosis capacity, a significantly reduced generation of reactive oxygen species, and reduced mRNA expression of CXCL8, CXCL1 and IL-1? after LPS stimulation. Based on IL-1? secretion after LPS/ATP stimulation, the inflammasome could be activated in cM and intM, but not in ncM. IFN? increased the expression of CD16 selectively on cM and induced a shift from cM into intM in vitro. In summary, bovine CD172a-positive mononuclear cells define three monocyte subsets with distinct phenotypic and functional differences. Bovine cM and intM share homologies with their human counterparts, whereas bovine ncM are not inflammatory monocytes. PMID:23967219

Hussen, Jamal; Düvel, Anna; Sandra, Olivier; Smith, David; Sheldon, Iain Martin; Zieger, Peter; Schuberth, Hans-Joachim



Monocyte transplantation for neural and cardiovascular ischemia repair.  


Neovascularization is an integral process of inflammatory reactions and subsequent repair cascades in tissue injury. Monocytes/macrophages play a key role in the inflammatory process including angiogenesis as well as the defence mechanisms by exerting microbicidal and immunomodulatory activity. Current studies have demonstrated that recruited monocytes/macrophages aid in regulating angiogenesis in ischemic tissue, tumours and chronic inflammation. In terms of neovascularization followed by tissue regeneration, monocytes/macrophages should be highly attractive for cell-based therapy compared to any other stem cells due to their considerable advantages: non-oncogenic, non-teratogenic, multiple secretary functions including pro-angiogenic and growth factors, straightforward cell harvesting procedure and non-existent ethical controversy. In addition to adult origins such as bone marrow or peripheral blood, umbilical cord blood (UCB) can be a potential source for autologous or allogeneic monocytes/macrophages. Especially, UCB monocytes should be considered as the first candidate owing to their feasibility, low immune rejection and multiple characteristic advantages such as their anti-inflammatory properties by virtue of their unique immune and inflammatory immaturity, and their pro-angiogenic ability. In this review, we present general characteristics and potential of monocytes/macrophages for cell-based therapy, especially focusing on neovascularization and UCB-derived monocytes. PMID:19754667

Sanberg, Paul R; Park, Dong-Hyuk; Kuzmin-Nichols, Nicole; Cruz, Eduardo; Hossne, Nelson Americo; Buffolo, Enio; Willing, Alison E



Studies of human antiviral CD8+ lymphocytes using class I peptide tetramers.  


Understanding the interactions between a host and a pathogen relies crucially on quantitative measurements of immune responses. Until recently, measurements of the levels of cellular immune responses, i.e. those mediated by CD4+ and CD8+ T lymphocytes have depended largely on culture in vitro and subsequent measurement of specific functions (such as cytolysis). More recently, new technologies based around tetrameric class I peptide complexes (tetramers) have allowed immunologists to measure CD8+ T lymphocyte levels directly ex vivo and independently of function. Since CD8+ lymphocytes play a key role in a number of important human viral infections, these tools have yielded useful insights into the dynamics, phenotype and function of human antiviral lymphocyte populations. In this review we describe some of the basic aspects of the biology of virus-specific CD8+ lymphocytes, and the current methods available to detect them. The use of tetramers has, in just four years, transformed our understanding of the immune responses against HIV, HTLV-1, HBV, HCV, CMV and EBV, and holds promise in a number of areas where quantitative analysis of the antiviral response in terms of both number and function is critical. PMID:11241799

Lechner, F; Cuero, A L; Kantzanou, M; Klenerman, P


Cyto-protective and immunomodulating properties of Amla ( Emblica officinalis) on lymphocytes: an in-vitro study  

Microsoft Academic Search

The fruits extracts of Emblica officinalis (Amla) has been reported to have strong anti-oxidant properties. There is a paucity of studies on the immunomodulatory properties of fruit extracts of Amla in immuno-compromised states, with the emphasis on lymphocytes. Therefore, the aim of the study was to determine the anti-oxidant and immunomodulatory properties of Amla using chromium (VI) as an immunosuppressive

M Sai Ram; D Neetu; B Yogesh; B Anju; P Dipti; T Pauline; S. K Sharma; S. K. S Sarada; G Ilavazhagan; Devendra Kumar; W Selvamurthy



A study on the beryllium lymphocyte transformation test and the beryllium levels in working environment.  


The relationship between airborne concentration of beryllium in the working environment and workers' beryllium lymphocyte transformation test (Be-LTT) values was examined based on data obtained from a four-year survey (1992-1995) conducted at beryllium-copper alloy manufacturing factories. This study showed that the T cells of workers continuously exposed to beryllium of more than 0.01 microgram/m3 could be activated and that the cell-mediated immune response of workers could be promoted. On the other hand, the Be-LTT of workers exposed to beryllium levels of less than 0.01 microgram/m3 was shown to be unaffected by beryllium. These findings suggest that beryllium sensitization is not manifested when level of beryllium in working environment are less than 0.01 microgram/m3. Therefore, in such cases workers do not develop Chronic beryllium disease (CBD). We concluded that the Be-LTT can be applied as a medical indicator to detect the development of CBD. PMID:9248221

Yoshida, T; Shima, S; Nagaoka, K; Taniwaki, H; Wada, A; Kurita, H; Morita, K



Studying the replication history of human B lymphocytes by real-time quantitative (RQ)-PCR.  


The cells of the adaptive immune system, B and T lymphocytes, each generate a unique antigen receptor through V(D)J recombination of their immunoglobulin (Ig) and T cell receptor (TCR) loci, respectively. Such rearrangements join coding elements to form a coding joint and delete the intervening DNA as circular excision products containing the signal joint. These excision circles are stable structures that cannot replicate and have no function in the cell. Since the coding joint in the genome is replicated with each cell division, the ratio between coding joints and signal joints in a population of B cells can be used as a measure for proliferation. This chapter describes a real-time quantitative (RQ-)PCR-based approach to quantify proliferation through calculating the ratio between coding joints and signal joints of the frequently occurring intronRSS-Kde rearrangements in the IGK light chain locus. The approach is useful to study basic B-cell biology as well as abnormal proliferation in human diseases. PMID:23296960

van Zelm, Menno C; Berkowska, Magdalena A; van Dongen, Jacques J M



Peripheral monocytes from diabetic patients with coronary artery disease display increased bFGF and VEGF mRNA expression  

PubMed Central

Background Macrophages can produce vascular endothelial growth factor (VEGF) in response to hypoxia, transforming growth factor ?1 (TGF-?1), angiotensin II, basic fibroblast growth factor (bFGF), and interleukin-1. These factors have been found in the serum of coronary artery disease (CAD) patients as well as in atherosclerotic lesions. The aim of the present study was to test the hypothesis that the expression of VEGF, TGF-?1 and bFGF in peripheral monocytes and lymphocytes is related to CAD. Methods Human Mononuclear cells and lymphocytes from peripheral blood were isolated from 53 donors undergoing angiography. Seventeen were found to be healthy and 36 were diagnosed with CAD. The respective mRNAs were extracted and quantified. Results The statistical analysis revealed a significant increase of the basal level expression for macrophage VEGF and bFGF in the CAD SA (stable angina) patient group compared to the noCAD (control) (p = 0.041 and p = 0.022 respectively) and CAD UA (unstable angina) (p = 0.024 and p = 0.005 respectively) groups, which was highly dependent on the diabetic status of the population. Furthermore, we demonstrated with an in vitro cell culture model that the levels of VEGF and bFGF in monocytes of healthy donors are not affected by short term exposure to increased glucose levels (usually observed in the diabetic patients) and/or statin. Conclusion Our findings display a statistically significant association of the increased VEGF and bFGF levels in peripheral monocytes, with stable angina and diabetes in coronary artery disease. The results give new insight to CAD and the impaired collateral vessel formation in diabetics.

Panutsopulos, Dimitrios; Zafiropoulos, Alexandros; Krambovitis, Elias; Kochiadakis, George E; Igoumenidis, Nikos E; Spandidos, Demetrios A



Immunological characterization and transcription profiling of peripheral blood (PB) monocytes in children with autism spectrum disorders (ASD) and specific polysaccharide antibody deficiency (SPAD): case study  

PubMed Central

Introduction There exists a small subset of children with autism spectrum disorders (ASD) characterized by fluctuating behavioral symptoms and cognitive skills following immune insults. Some of these children also exhibit specific polysaccharide antibody deficiency (SPAD), resulting in frequent infection caused by encapsulated organisms, and they often require supplemental intravenous immunoglobulin (IVIG) (ASD/SPAD). This study assessed whether these ASD/SPAD children have distinct immunological findings in comparison with ASD/non-SPAD or non-ASD/SPAD children. Case description We describe 8 ASD/SPAD children with worsening behavioral symptoms/cognitive skills that are triggered by immune insults. These ASD/SPAD children exhibited delayed type food allergy (5/8), treatment-resistant seizure disorders (4/8), and chronic gastrointestinal (GI) symptoms (5/8) at high frequencies. Control subjects included ASD children without SPAD (N = 39), normal controls (N = 37), and non-ASD children with SPAD (N = 12). Discussion and Evaluation We assessed their innate and adaptive immune responses, by measuring the production of pro-inflammatory and counter-regulatory cytokines by peripheral blood mononuclear cells (PBMCs) in responses to agonists of toll like receptors (TLR), stimuli of innate immunity, and T cell stimulants. Transcription profiling of PB monocytes was also assessed. ASD/SPAD PBMCs produced less proinflammatory cytokines with agonists of TLR7/8 (IL-6, IL-23), TLR2/6 (IL-6), TLR4 (IL-12p40), and without stimuli (IL-1ß, IL-6, and TNF-?) than normal controls. In addition, cytokine production of ASD/SPAD PBMCs in response to T cell mitogens (IFN-?, IL-17, and IL-12p40) and candida antigen (Ag) (IL-10, IL-12p40) were less than normal controls. ASD/non-SPAD PBMDs revealed similar results as normal controls, while non-ASD/SPAD PBMCs revealed lower production of IL-6, IL-10 and IL-23 with a TLR4 agonist. Only common features observed between ASD/SPAD and non-ASD/SPAD children is lower IL-10 production in the absence of stimuli. Transcription profiling of PB monocytes revealed over a 2-fold up (830 and 1250) and down (653 and 1235) regulation of genes in ASD/SPAD children, as compared to normal (N = 26) and ASD/non-SPAD (N = 29) controls, respectively. Enriched gene expression of TGFBR (p < 0.005), Notch (p < 0.01), and EGFR1 (p < 0.02) pathways was found in the ASD/SPAD monocytes as compared to ASD/non-SPAD controls. Conclusions The Immunological findings in the ASD/SPAD children who exhibit fluctuating behavioral symptoms and cognitive skills cannot be solely attributed to SPAD. Instead, these findings may be more specific for ASD/SPAD children with the above-described clinical characteristics, indicating a possible role of these immune abnormalities in their neuropsychiatric symptoms.



[Clinical usefulness of the immunophenotypic study of circulating lymphocytes in leukemic lymphoproliferative diseases].  


The immunophenotypes (IF) on peripheral lymphocytes of 24 B-CLL in different stage, 2 PLL and 14 leukemic B-non Hodgkin lymphomas were investigated. As regard to B-CLL and PLL, the results are similar to those reported so far. In stage A and B of B-CLL the IF appear less variable than in advanced stages where a decrease of CD21+ and an increase of both CD25+ and CD38+ lymphocytes were observed. In the lymphocytic, small cleaved cell lymphomas and splenic lymphomas, the peripheral IF correspond to the theoretical ones of respective lymphoma tissues. On the contrary they disagree in three cases of large cells, mixed small and cleaved cell, immunoblastic lymphomas. These features are discussed. PMID:7567166

Büchi, G; Messina, M; Girotto, M; Termine, G; Morello, M; Orlassino, R; Autino, R; Grosso, E



Human brucellosis is characterized by an intense Th1 profile associated with a defective monocyte function.  


In animal models, a defective Th1 response appears to be critical in the pathogenesis of brucellosis, but the Th1 response in human brucellosis patients remains partially undefined. Peripheral blood from 24 brucellosis patients was studied before and 45 days after antibiotherapy. Twenty-four sex- and age-matched healthy donors were analyzed in parallel. Significantly increased levels of interleukin 1beta (IL-1beta), IL-2, IL-4, IL-6, IL-12p40, gamma interferon (IFN-gamma), and tumor necrosis factor alpha (TNF-alpha), but not of IL-10, in serum and/or significantly increased percentages of samples with detectable levels of these cytokines, measured by enzyme-linked immunosorbent assays (ELISA), were found for untreated brucellosis patients, but these levels were reduced and/or normalized after treatment. Flow cytometry studies showed that the intracytoplasmic expression of IFN-gamma, IL-2, and TNF-alpha, but not that of IL-4, by phorbol myristate-activated CD4(+) CD3(+) and CD8(+) CD3(+) T lymphocytes was significantly increased in untreated brucellosis patients and was also partially normalized after antibiotherapy. The percentage of phagocytic cells, the mean phagocytic activity per cell, and the phagocytic indices for monocytes at baseline were defective and had only partially reverted at follow-up. T lymphocytes from untreated brucellosis patients are activated in vivo and show Th1 cytokine production polarization, with strikingly high serum IFN-gamma levels. In spite of this Th1 environment, we found deficient effector phagocytic activity in peripheral blood monocytes. PMID:20404074

Rodríguez-Zapata, Manuel; Matías, Marlene J; Prieto, Alfredo; Jonde, Marco A; Monserrat, Jorge; Sánchez, Lorenzo; Reyes, Eduardo; De la Hera, Antonio; Alvarez-Mon, Melchor



A comparative study of lymphocytes in effusions of patients with tuberculosis or malignant disease.  


The diagnostic value of the determination of the relative distribution of B (SmIg-positive) and T (E rosetting) cells in the blood and in pleural or ascitic effusions was evaluated in 10 patients with tuberculosis and in six with malignant diseases. A higher percentage of T and a lower percentage of B lymphocytes in serous fluids than in peripheral blood was observed for both groups of patients. We conclude that this procedure is valueless in differentiating between tuberculosis and malignant diseases when effusions containing predominantly lymphocytes and devoid of metastatic cells are examined. PMID:6975680

Falcão, R P; Bottura, C



A comparative study of lymphocytes in effusions of patients with tuberculosis or malignant disease.  

PubMed Central

The diagnostic value of the determination of the relative distribution of B (SmIg-positive) and T (E rosetting) cells in the blood and in pleural or ascitic effusions was evaluated in 10 patients with tuberculosis and in six with malignant diseases. A higher percentage of T and a lower percentage of B lymphocytes in serous fluids than in peripheral blood was observed for both groups of patients. We conclude that this procedure is valueless in differentiating between tuberculosis and malignant diseases when effusions containing predominantly lymphocytes and devoid of metastatic cells are examined.

Falcao, R P; Bottura, C



Monocyte/macrophage proteomics: recent findings and biomedical applications.  


Macrophages, originating from the migration and differentiation of circulating monocytes into virtually all tissues, are extremely flexible and plastic cells that play vital homeostatic roles, but also contribute to the pathophysiology of many human diseases. For these reasons, they are intensively studied by different approaches, recently including proteomics. Macrophage cells can be taken from a range of different sources, including blood monocytes and macrophages from tissues. Macrophages can also be generated by in vitro culture from blood monocytes, and cell lines derived from this lineage can be used. Similarly, many different proteomic techniques can be used, ranging from classic approaches based on 2D gel electrophoresis to more recent high-throughput gel-free techniques essentially based on mass spectrometry. Here, we review the application of such techniques to the study of monocytes/macrophages, and summarize some results potentially relevant to two paradigmatic conditions - atherosclerosis and disorders of iron metabolism. PMID:22462790

Castagna, Annalisa; Polati, Rita; Bossi, Alessandra Maria; Girelli, Domenico



Study on the blood-borne virus co-infection and T lymphocyte subset among intravenous drug users  

Microsoft Academic Search

AIM: To investigate the features of various blood- borne virus infections and co-infection in intravenous drug users (IDUs), and to examine the correlation of T lymphocyte subsets with virus co-infection. METHODS: Four hundred and six IDUs without any clinical manifestation of hepatitis and 102 healthy persons were enrolled in this study. HBV-DNA and HCV-RNA were detected by fluorescence quantitative PCR.

Jian-Rong Li; Rui-Yu Gong; Kun-Lun Tian; Jing Wang; Yi-Xin Wang; Han-Ju Huang



Effect of an Intramuscular Injection of Human Leukocyte Interferon on Blood Leukocyte Counts and Proportions of Lymphocytes Forming E, EA and EAC Rosettes  

Microsoft Academic Search

Following an intramuscular injection of human leukocyte interferon blood leukocyte counts decreased in most patients as tested 24 h after the injection. This decrease comprised lymphocytes, monocytes and granulocytes. 24 h after interferon injection, the proportion of lymphocytes forming E rosettes was increased in most patients, whereas the proportion of lymphocytes forming EA and EAC rosettes was not significantly changed.Copyright

Stefan Einhorn; Henric Blomgren; Hans Strander



Monocytes from sickle cell disease patients induce differential pulmonary endothelial gene expression via activation of NF-?B signaling pathway.  


Monocyte-endothelial interactions play an important role in inflammatory diseases and may modulate vasculopathy in sickle cell disease, a disorder with an important inflammatory component. We co-incubated normal and sickle monocytes, lymphocytes and TNF-? with pulmonary microvascular and arterial endothelial cells and compared the expression of genes coding for adhesion molecules and cytokines that might contribute to sickle vasoocclusion. Monocyte-endothelial cell co-incubation resulted in up-regulation of L-selectin, E-selectin, VCAM-1, ICAM-1, MCP-1, MMP-1, TNF-?, IL-6 and IL-1? and down-regulation of eNOS. Lymphocyte-endothelial cell co-incubations, induced similar effects restricted to pulmonary artery endothelial cells. TNF-? had similar effects on the endothelial cells as monocytes did, however monocyte induced gene expression in endothelial cells was not TNF-? dependent but was regulated through the NF-?B pathway. Sickle monocytes lead to altered expression of L-selectin, MCP-1 and MMP-1 in pulmonary vascular endothelium when compared with normal monocytes. The gene expression changes we observed could reflect pathological events of sickle vasoocclusion. PMID:22264835

Safaya, Surinder; Steinberg, Martin H; Klings, Elizabeth S



Ovine lentivirus is macrophagetropic and does not replicate productively in T lymphocytes.  

PubMed Central

The lentiviruses of sheep, goats, and horses cause chronic multiorgan disease in which macrophages are highly permissive for viral replication. Monocytes, which mature into macrophages, are thought to be latently infected with lentivirus, but the extent to which other leukocytes are infected is unknown. Dendritic cells have not been studied separately from monocytes and T-cell subsets have not been examined in previous attempts to identify infected cells in peripheral blood mononuclear cells (PBMC). We found no evidence of T-cell tropism using an animal-passaged, pathogenic ovine lentivirus. Phytohemagglutinin-stimulated infectious PBMC produced 20-fold less virus than differentiated macrophages, and cocultivation of infectious PBMC with fresh, uninfected phytohemagglutinin blasts did not facilitate virus replication. Furthermore, central lymph cells, the best in vivo source of purified lymphocytes, lacked virus and did not yield virus upon in vitro cultivation. In contrast, cultivated blood-derived macrophages were highly permissive for viral replication. To identify the latently infected PBMC, PBMC from infected sheep were selectively depleted of monocytes and B cells by passage over nylon wool and then of nonadherent cells bearing CD4, CD8, T19, gamma delta T-cell receptor, CD45RA, or major histocompatibility complex class II antigens by panning. Removal of adherent monocytes and B cells or of adherent cells and the three major T-cell subsets (CD4+, CD8+, T19+) did not decrease the infectivity of PBMC. The richest sources of infected cells in fresh PBMC were CD45RA+ and major histocompatibility complex class II+ nonadherent cells, which are three characteristics of dendritic cells. Thus, the dendritic cell, and not the monocyte or the CD4+ cell, is probably the predominant infected cell type in blood. Images

Gorrell, M D; Brandon, M R; Sheffer, D; Adams, R J; Narayan, O



X-linked B-lymphocyte immune defect in CBA/HN mice. I. Studies of the function and composition of spleen cells  

PubMed Central

A study of the composition and functional properties of spleen cells from the immune deficient CBA/HN mice and their F1 progeny is reported. While abnormalities were seen in both the numbers and function of thymus-independent (B) lymphocytes, all studies involving thymus- dependent (T) lymphocytes were normal. The X-linked nature of the immune defect in these mice was therefore attributed to abnormal or absent B lymphocytes. The possible nature of this defect and the similarity of the immune defect in these mice to certain human X-linked immunodeficiency diseases are discussed.



Interaction of pseudorabies virus with porcine peripheral blood lymphocytes  

Microsoft Academic Search

To examine effects of pseudorabies virus (PrV) on immune cells, we investigated the ability of PrY to in- fect and replicate in porcine peripheral blood leukocytes (PBLs). Flow cytometric analysis revealed a leukopenia after challenge, with loss of 40% of circulating monocytes and 50% of circulating lymphocytes. Virus was isolated from PBLs of challenged pigs by cocultivation with por- cine

Gregory R. Page; Fun-In Wang; Edwin C. Hahn



Microsoft Academic Search

It has been recognized for more than half a century that, in addition to occasional mammary gland alveolar or ductal epithelial cell fragments, colos- trum and milk consistently contain significant concentrations of viable leuko- cytes. These include lipid-laden macrophages that can exhibit ameboid and phagocytic activity, polymorphonuclear neutrophils, and lymphocytes of various sizes (1, 2). The overall concentration of these



Immunologic and Immunohistochemical Studies on Chronic Lymphocytic Thyroiditis with or without Thyroid Lymphoma  

Microsoft Academic Search

The local immunologic phenomena in the thyroid gland of 16 patients with chronic lymphocytic thyroiditis (CLTH) were investigated; 5 of these cases were associated with thyroid non-Hodgkin’s lymphomas (NHL). All patients were admitted because of struma, growing slowly in patients with CLTH alone and rapidly in those with associated thyroid NHL. CLTH was confirmed by histologic findings, including the presence

Katsuyuki Aozasa; Kazuo Tajima; Nobuhiko Tominaga; Shuichi Katagiri; Takeshi Yonezawa; Fumio Matsuzuka; Kanji Kuma; Masuomi Sawada



Glucose Regulates Monocyte Adhesion Through Endothelial Production of Interleukin8  

Microsoft Academic Search

We have shown that glucose increases monocyte adhesion to human aortic endothelial cells (HAECs) in vitro.1 In the present study, we examined mechanisms by which glucose stimulates monocyte:endothelial interactions. HAECs cultured for 7 days in 25 mmol\\/L glucose had a 2-fold elevation in interleukin-8 (IL-8) secretion over control cells cultured in 5.5 mmol\\/L glucose (P0.001). Use of a neutralizing antibody

Suseela Srinivasan; Michael Yeh; Eric C. Danziger; Melissa E. Hatley; Anna E. Riggan; Norbert Leitinger; Judith A. Berliner; Catherine C. Hedrick



A study of the Interaction Between Cetirizine and Plasma Membrane of Eosinophils, Neutrophils, Platelets and Lymphocytes using A fluorescence Technique  

PubMed Central

The effect of cetirizine on plasma membrane fluidity and heterogeneity of human eosinophils, neutrophils, platelets and lymphocytes was investigated using a fluorescence technique. Membrane fluidity and heterogeneity were studied by measuring the steady-state fluorescence anisotropy and fluorescence decay of 1-(4- trimethylammonium-phenyl)-6-phenyl-1, 3, 5-hexatriene (TMA-DPH) incorporated in the membrane. The results demonstrate that cetirizine (1 ?g/ml) induced a significant increase in the Hpid order in the exterior part of the membrane and a decrease in membrane heterogeneity in eosinophils, neutrophils and platelets. Moreover, cetirizine blocked the PAF induced changes in membrane fluidity in these cells. Cetirizine did not influence significantly the plasma membrane of lymphocytes. These data may partially explain the effect ofcetirizine on inflammatory cell activities.

Oggiano, N.; Giorgi, P. L.; Rihoux, J-P.



T helper type 1 lymphocytes drive inflammation in human atherosclerotic lesions.  


Atherosclerotic lesions are infiltrated by macrophages and T lymphocytes, potentially reactive to pathogens. We studied in vivo activated T lymphocytes that infiltrate atherosclerotic plaques of Helicobacter pylori-infected patients with or without anti-Chlamydia pneumoniae antibodies. In all atherosclerotic lesions, T helper type 1 (Th1) cells were predominant. C. pneumoniae-specific T cells were detected only in the plaques of anti-C. pneumoniae seropositive patients, whereas H. pylori-specific T cells were found in the gastric mucosa but not in the plaques of the same patients. Plaque-derived Th1 cells expressed cytotoxicity, proapoptotic activity, and help for monocyte tissue factor production. Although multifactorial, atherosclerosis can be regarded as a Th1-driven immunopathological condition. PMID:12740434

Benagiano, Marisa; Azzurri, Annalisa; Ciervo, Alessandra; Amedei, Amedeo; Tamburini, Carlo; Ferrari, Mauro; Telford, John L; Baldari, Cosima T; Romagnani, Sergio; Cassone, Antonio; D'Elios, Mario M; Del Prete, Gianfranco



Study of T-lymphocyte subsets of healthy and Mycobacterium avium subsp. paratuberculosis-infected cattle.  

PubMed Central

The relative contributions of T-lymphocyte subsets to host defense in cattle infected with Mycobacterium avium subsp. paratuberculosis is reported. The subsets were purified with appropriate monoclonal antibodies and a magnetic bead column separation system, and their purity was verified by flow cytometry. Biological activity of each subset, expressed as lymphoproliferation and gamma interferon (IFN-gamma) production, was measured in response to phytohemagglutinin (PHA) and an M. avium antigen preparation (A-PPD). IFN-gamma was measured by antibody capture enzyme-linked immunosorbent assay. The results showed a correlation between proliferation and IFN-gamma production in response to A-PPD but not to PHA. In response to PHA, CD4+ lymphocytes were the most prolific producers of IFN-gamma. CD8+ lymphocytes produced IFN-gamma to a lesser extent, whereas gammadelta+ T lymphocytes produced little or no IFN-gamma. Differences observed between the amount of IFN-gamma produced by CD4+ versus CD8+ cells and CD4+ versus gammadelta+ cells were significant (P < 0.01), but those between peripheral blood mononuclear cells (PBMC) and CD4+ T cells were not. Similar responses to A-PPD were observed except that PBMC produced higher levels of IFN-gamma than did CD4+ T cells. These data for cattle are similar to observations made for other animal species, where CD4+ cells are the major type of T lymphocytes producing IFN-gamma. They further suggest that whatever the role gammadelta+ T cells may play in paratuberculosis, it is not likely to be mediated by IFN-gamma production.

Bassey, E O; Collins, M T



DNA damage and repair measurements from cryopreserved lymphocytes without cell culture--a reproducible assay for intervention studies.  


Single-cell gel electrophoresis (the Comet assay) can be used to measure DNA damage and DNA repair capacity (DRC). However, to test DRC of cryopreserved lymphocytes, published methods include steps for cell culturing and phytohemagglutinin stimulation, which may limit use of this assay in intervention studies. We developed a modified Comet assay protocol that allows us to measure DRC from cryopreserved lymphocytes without these in vitro manipulations. Assay reproducibility was evaluated by performing the assay six times on different dates using six aliquots from one blood draw of one individual. The interindividual variation was assessed by performing the assay using one aliquot from six individuals. When gamma-irradiation was used as the mutagen, intra-assay coefficients of variation (CVs.) for baseline DNA damage, damage after gamma-irradiation exposure, and DRC--measured as tail moment--were 8, 31, and 10%, respectively. Interindividual CVs. were higher. When H(2)O(2) was used as the mutagen, intra-assay CVs. for damage measurements were lower for a protocol modification that included damage and repair at 37 degrees C (CVs. ranging from 8 to 35%) than for the more standard 4 degrees C protocol. Analyzing moment arm--the average distance of DNA migration within the tail--yielded similar results. DNA repair was successfully detected in each experiment. Comparing freshly isolated lymphocytes to cryopreserved lymphocytes from the same individuals' blood draw indicated that DRC was highly correlated when determined using moment arm values. This modified protocol extends the use of the Comet assay to measuring DRC in intervention studies (e.g., dietary interventions) in that it assesses cellular response after cryopreservation without cell culture or other extensive manipulation. PMID:16673412

Chang, Jyh-Lurn; Chen, Gang; Lampe, Johanna W; Ulrich, Cornelia M



Honey stimulates inflammatory cytokine production from monocytes  

Microsoft Academic Search

Clinical observations indicate that honey may initiate or accelerate the healing of chronic wounds and has, therefore, been claimed to have anti-inflammatory properties. The aim of this study was to investigate the effects of honey on the activation state of immunocompetent cells, using the monocytic cell line, MonoMac-6 (MM6), as a model.We investigated the effect of each of the three

A. J Tonks; R. A Cooper; K. P Jones; S Blair; J Parton



Monoclonal antibodies to antigens on human neutrophils, activated T lymphocytes, and acute leukemia blast cells  

SciTech Connect

The authors describe the production of two mouse hybridomas secreting monoclonal antibodies to antigenic determinants of the surface membranes of human neutrophils, activated T lymphocytes, and acute leukemic blast cells. The degree of lymphocyte stimulation was estimated from incorporation of /sup 3/H-thymidine with parallel microculture. Monoclonal antibodies of supernatants of hybridoma cultures shown here reacted in both immunofluorescence test and cytotoxicity test with surface membrane antigens on the majority of neutrophils and PHA-activated peripheral blood lymphocytes from healthy subjects, but did not give positive reactions with unactivated lymphocytes, adherent monocytes, erythrocytes, and alloantigen-stimulated lymphocytes.

Miterev, G.Yu.; Burova, G.F.; Puzhitskaya, M.S.; Danilevich, S.V.; Bulycheva, T.I.



Genome-wide association study for T lymphocyte subpopulations in swine  

PubMed Central

Background Lymphocytes act as a major component of the adaptive immune system, taking very crucial responsibility for immunity. Differences in proportions of T-cell subpopulations in peripheral blood among individuals under same conditions provide evidence of genetic control on these traits, but little is known about the genetic mechanism of them, especially in swine. Identification of the genetic control on these variants may help the genetic improvement of immune capacity through selection. Results To identify genomic regions responsible for these immune traits in swine, a genome-wide association study was conducted. A total of 675 pigs of three breeds were involved in the study. At 21 days of age, all individuals were vaccinated with modified live classical swine fever vaccine. Blood samples were collected when the piglets were 20 and 35 days of age, respectively. Seven traits, including the proportions of CD4+, CD8+, CD4+CD8+, CD4+CD8?, CD4?CD8+, CD4?CD8? and the ratio of CD4+ to CD8+ T cells were measured at the two ages. All the samples were genotyped for 62,163 single nucleotide polymorphisms (SNP) using the Illumina porcineSNP60k BeadChip. 40833 SNPs were selected after quality control for association tests between SNPs and each immune trait considered based on a single-locus regression model. To tackle the issue of multiple testing in GWAS, 10,000 permutations were performed to determine the chromosome-wise and genome-wise significance levels of association tests. In total, 61 SNPs with chromosome-wise significance level and 3 SNPs with genome-wise significance level were identified. 27 significant SNPs were located within the immune-related QTL regions reported in previous studies. Furthermore, several significant SNPs fell into the regions harboring known immunity-related genes, 14 of them fell into the regions which harbor some known T cell-related genes. Conclusions Our study demonstrated that genome-wide association studies would be a feasible way for revealing the potential genetics variants affecting T-cell subpopulations. Results herein lay a preliminary foundation for further identifying the causal mutations underlying swine immune capacity in follow-up studies.



Anti-human IG and B lymphocytes reconstitute the proliferative response to Con A of T lymphocytes depleted of accessory cells  

SciTech Connect

The requirements for the induction of proliferation of human peripheral blood mononuclear cells (PBM) by low concentrations of Concanavalin A (Con A) were studied using /sup 3/H-thymidine incorporation to assay DNA synthesis. Removal of plastic-adherent cells from PBM reduced the proliferative response of lymphocytes to Con A by 80%. Passage of these cells over nylon wool columns totally eliminated their response to Con A. Addition of adherent monocytes or rabbit anti-human IgG conjugated to polyacrylamide beads (anti-IgG beads) to the non-adherent lymphocyte population reconstituted the proliferative response. The lymphoblasts activated in these cultures were more than 90% T11 positive. Anti-IgG beads did not activate lymphocyte proliferation in the absence of Con A. The response of non-adherent lymphocytes to Con A could also be reconstituted by unconjugated rabbit anti-human IgG antiserium. Purified T cells could only be activated by anti-IgG beads and low concentrations of Con A in the presence of irradiated B cells. These results suggest that /sup 3/H-thymidine incorporation by T cells induced by anti-IgG beads and a low concentration of Con A depends on the interaction of anti-IgG and B cells. The authors suggest that B cell produce growth factors for T cells after interacting with anti-IgG.

Tsuda, T.; Kim, Y.T.; Schwab, R.; Siskind, G.W.; Weksler, M.E.



Extracellular adenine nucleotides inhibit the release of major monocyte recruiters by human monocyte-derived dendritic cells  

Microsoft Academic Search

Extracellular ATP is known to affect the maturation of monocyte-derived dendritic cells mainly by regulation of cytokines and costimulatory molecules. The present study describes the inhibition of MCP-1 (CCL2) and MIP-1? (CCL3) release by human monocyte-derived dendritic cells in response to adenine nucleotides. Our pharmacological data support the involvement of P2Y11 and P2Y1 purinergic receptors in the downregulation of these

Michael Horckmans; Brice Marcet; Frédéric Marteau; Frédéric Bulté; Arielle Maho; Marc Parmentier; Jean-Marie Boeynaems; Didier Communi



Effects of vascular endothelial growth factor (VEGF) and chondroitin sulfate A on human monocytic THP1 cell migration  

Microsoft Academic Search

Angiogenesis serves as a crucial factor in disease development and progression, such as cancer metastasis, and monocyte migration is one of the key steps for angiogenesis. Therapeutic modulation of angiogenesis is a promising new therapeutic avenue under investigation. In this study, effects of vascular endothelial growth factor (VEGF) and chondroitin sulfate A on monocyte migration were investigated. Human monocytic THP-1

Y. Liu; H. Yang; K. Otaka; H. Takatsuki; A. Sakanishi



Extracellular adenine nucleotides inhibit the release of major monocyte recruiters by human monocyte-derived dendritic cells.  


Extracellular ATP is known to affect the maturation of monocyte-derived dendritic cells mainly by regulation of cytokines and costimulatory molecules. The present study describes the inhibition of MCP-1 (CCL2) and MIP-1alpha (CCL3) release by human monocyte-derived dendritic cells in response to adenine nucleotides. Our pharmacological data support the involvement of P2Y11 and P2Y1 purinergic receptors in the downregulation of these major monocyte recruiters. Migration assays have demonstrated that supernatants of dendritic cells treated with adenine nucleotides or anti-MCP-1/MIP-1alpha blocking antibodies display a strongly reduced capacity to attract monocytes and immature dendritic cells. PMID:16413542

Horckmans, Michael; Marcet, Brice; Marteau, Frédéric; Bulté, Frédéric; Maho, Arielle; Parmentier, Marc; Boeynaems, Jean-Marie; Communi, Didier



I. Suppression by compound 48\\/80 of bacterial endotoxin-inducible monocytic tissue factor activity: direct blockade of factor VII binding to THP1 monocytes 1 This study was presented in part in abstract form to the American Society for Biochemistry and Molecular Biology annual meeting held in San Francisco, CA, 16–20 May 1999. 1  

Microsoft Academic Search

Hypercoagulation with upregulated monocytic tissue factor (TF) activity often occurs under a variety of inflammatory conditions including endotoxemia. The antagonism to bacterial endotoxin (LPS) signaling often results in the depression in TF upregulation. We herein report that compound 48\\/80 (48\\/80) significantly depressed LPS-induced TF activity in human and cebus monkey peripheral blood monocytes. Employing a model monocyte-like cell line (THP-1),

Arthur J Chu; Melissa A Walton; Anne Seto; Melissa J Fox; Jai K Prasad; Zhen-Guo Wang



Characterization of substance P binding to human monocytes/macrophages.  

PubMed Central

Substance P (SP), a member of the tachykinin family of neuropeptides, can immunomodulate human T cells and monocytes. SP has been shown to stimulate human monocytes to produce inflammatory cytokines and superoxide ions, and it enhances tumoricidal activity in vitro. A specific SP receptor, however, has not been identified on human monocytes/macrophages. In this study, we report that 125I-SP binds to human monocytes/macrophages with high affinity and specificity (Kd = 2.7 x 10(-8) to 5.5 x 10(-8) M). Our measurements of binding affinity to this single class of receptors were possible only when experiments were performed in the presence of excess serine proteinase inhibitor (serpin) enzyme complex receptor ligand. We determined that 125I-SP bound to a specific receptor on human monocytes/macrophages and that this binding was detectable as early as 6 h and was maintained throughout 6 to 8 weeks in culture. Modulation of the diverse immunological and inflammatory effects of SP on human monocytes may be mediated through this specific SP receptor. Images

Lucey, D R; Novak, J M; Polonis, V R; Liu, Y; Gartner, S



Oxidized Low Density Lipoprotein Induces Differentiation and Adhesion of Human Monocytes and the Monocytic Cell Line U937  

NASA Astrophysics Data System (ADS)

Hypercholesterolemia is a major risk factor for development of atherosclerosis. In experimental animals fed a high-cholesterol diet, monocytes adhere to the arterial endothelium and penetrate into the intima where they differentiate into macrophages and ingest lipids thus giving rise to fatty streaks, the earliest type of atherosclerotic plaque. Macrophages express few receptors for normal low density lipo-protein (LDL) but can take up oxidized LDL by way of a scavenger receptor. The present study was designed to investigate the possible role of oxidized LDL in recruitment of resident intimal macrophages. We found that oxidized LDL induced enhanced expression of major histocompatibility complex class II molecules on human monocytes and U937 cells, a well-established system for studies of monocytic differentiation. Oxidized LDL also induced enhanced expression of the surface antigen LEuM3 but caused decreased expression of CD4 antigen, a pattern compatible with expression of a more differentiated macrophage-like phenotype. Oxidized LDL also initiated aggregation of monocytes and U937 cells and stimulated adhesion of U937 cells to cultured endothelial cells. The results indicate that oxidized LDL may contribute to development of atherosclerosis by inducing adhesion of monocytes to the arterial intima and by stimulating intimal monocytes to differentiate into resident macrophages.

Frostegard, Johan; Nilsson, Jan; Haegerstrand, Anders; Hamsten, Anders; Wigzell, Hans; Gidlund, Magnus



Field Studies of Cytotoxic T Lymphocytes in Malaria Infections: Implications for Malaria Vaccine Development  

Microsoft Academic Search

The search for a cytotoxic T lymphocyte (CTL)-inducing malaria vaccine has moved forward from epitope identification to planning stages of safety and immunogenicity trials of candidate vaccines. Development of CTL-inducing vaccine candidates has taken center stage based on the observation that CTL-mediated protection might be the dominant mechanism by which sterile immunity is achieved in irradiated sporozoite immunization experiments in

Michael Aidoo; Venkatachalam Udhayakumar



T-cell-mediated and antigen-dependent differentiation of human monocyte into different dendritic cell subsets: a feedback control of Th1/Th2 responses.  


It is well established that human monocytes differentiate into dendritic cells (DCs) when cultured with certain cytokine cocktails, such as granulocyte-macrophage colony-stimulating factor and interleukin-4. Conversely, it is not completely established which cell population synthesizes the cytokines required for monocyte differentiation and how their secretion is regulated. We show that on specific activation T cells induce the differentiation into DCs of antigen-presenting and bystander monocytes. Monocytes exposed to cytokines released by Th1 and Th0 lymphocytes differentiate into DCs with a reduced antigen uptake and antigen presentation capacity. Moreover, these DCs show a limited capacity to induce Th1 polarization of naive T cells but are capable of priming interleukin-10-secreting T cells. Conversely, DCs derived from monocytes sensing cytokines released by Th2 lymphocytes are antigen-presenting-cell (APC) endowed with a marked Th1 polarization capacity. Monocytes are corecruited with lymphocytes in chronic inflammation sites; thus our results suggest that functionally different DCs can be generated in environments characterized by the prevalent release of Th1-, Th0-, or Th2-associated cytokines. Because the APC capacities of these DCs have opposite functional consequences, a contribution in the regulation of the ongoing immune response by monocyte-derived inflammatory DCs is envisaged. PMID:18556459

Mariotti, Sabrina; Sargentini, Valeria; Marcantonio, Cinzia; Todero, Emiliano; Teloni, Raffaela; Gagliardi, Maria Cristina; Ciccaglione, Anna Rita; Nisini, Roberto



Studies on the interaction of the Sophora japonica lectin and concanavalin A with erythrocytes and lymphocytes.  

PubMed Central

The agglutinating activity of lectins from the seeds of Sophora japonica and Canavalia ensiformis (concanavalin A) with human and murine erythrocytes and lymphocytes have been compared to one another and related to the mitogenic and immunosuppressive properties of these purified proteins. The S. japonica lectin, which demonstrates blood group specificity, is more active than concanavalin A with human erythrocytes, but has a much lower reactivity than concanavalin A with murine red blood cells. Ficin treatment of human erythrocytes results in an increase in agglutinability by both lectins as well as causing the appearance of S. japonica lectin receptors on type O cells. Treatment of murine reythrocytes with ficin alone or followed by beta-galactosidase causes the cells to be more reactive with concanavalin A. Beta-Galactosidase alone has no observable affect on the cells. In contrast, the agglutinability of cells by the S. japonica lectin increases after ficin treatment but is not affected by beta-galaetosidose treatment either after or in the absence of ficinization. Murine lymphocytes react with both lectins in a manner paralleling the agglutination patterns of murine erythrocytes. The S. japonica lectin appears to be devoid of mitogenic and immuno-suppressive activity, in contrast to concanavalin A which suppresses the T helper-dependent antibody response to sheep erythrocytes. These results are discussed in terms of the types of lectin receptors on lymphocytes related to agglutination, induction of blastogenesis and immuno-suppression.

Poretz, R D; Barth, R F



Model for the Control of Potassium Transport in PHA-Stimulated Human Blood Lymphocytes.  

National Technical Information Service (NTIS)

Results are reported for studies on the following: lymphocyte plasma-membrane ATP ase; substrate specificity of lymphocyte membrane phosphatase activity; effect of PHA on lymphocyte membrane ATP ase; correlation of K exp + with lymphocyte K exp + concentr...

G. B. Segel W. Simon M. A. Lichtman



The relative role of neutrophils and platelets in the local accumulation of circulating lymphocytes at sites of ionophore A23187 inoculation  

SciTech Connect

The early cellular infiltrate at inflammatory sites consists predominantly of neutrophils and cells of the monocyte/macrophage lineage. The mechanism by which circulating, unsensitized lymphocytes accumulate at sites of inflammation is unknown. The pattern of accumulation of 111indium-labeled circulating thymocytes in response to local injections of the ionophore A23187 was studied and compared with the pattern of (125)iodinated albumin accumulation as a measure of vascular permeability. The kinetics of thymocyte accumulation differed from those of vascular permeability. Sublethal total-body irradiation (750 rads) markedly decreased thymocyte accumulation but had little effect on vascular permeability. Irradiation of the local site alone had no effect. T lymphocyte, T lymphoblast, and platelet accumulation generally followed the same pattern as thymocytes. Intravenous injection of neutrophils, but not platelets, partially restored lymphocyte accumulation in vivo in irradiated mice via a pathway involving the circulating neutrophil, and seemed to be independent of changes in vascular permeability.

Hayes, J.M.; Simmons, R.L. (Univ. of Pittsburgh, PA (USA))



Human monocyte response to retrieved polymethylmethacrylate particles.  


The purpose of this study was to compare retrieved polymethylmethacrylate (PMMA) particles from failed total hip arthroplasties in terms of size, shape, and the response of human monocytes with commercially available particles. PMMA particles were isolated from peri-implant tissues of five failed cemented total hip arthroplasties using tissue digestion and a sucrose density gradient technique. Prepolymerized cement powder and those from which barium sulfate had been removed were examined for comparison. After exposure of peripheral human monocytes to PMMA particles, tumor necrosis factor-alpha and interleukin-6 in medium were measured by using enzyme-linked immunosorbent assays. Image analysis revealed that retrieved particles were larger (retrieved: 1.24 microm; prepolymerized cement powder: 0.83 microm; barium sulfate-free powder: 0.87 microm) and were more irregular in shape and rougher than commercially available particles. Cytokine release was increased by all PMMA particle species. However, commercially available PMMA particles stimulated the release of necrosis factor-alpha and interleukin-6 more strongly than did retrieved particles at very high doses. The observed difference in monocyte response might be due to the volume of the challenged particles. Another possible reason for the difference might be alteration of the surface chemistry of particles in situ and the difference in surface morphology between them. PMID:12209918

Miyaguchi, Masatsugu; Kobayashi, Akio; Iwaki, Hiroyoshi; Ohashi, Hirotsugu; Kadoya, Yoshinori; Yamano, Yoshiki



Caffeine sensitization of cultured mammalian cells and human lymphocytes irradiated with gamma rays and fast neutrons: a study of relative biological effectiveness in relation to cellular repair  

Microsoft Academic Search

The sensitizing effects of caffeine were studied in baby hamster kidney (BHK-21) cells and human lymphocytes following irradiation with gamma rays and fast neutrons. Caffeine sensitization occurred only when log-phase BHK cells and mitogen-stimulated lymphocytes were exposed to the two radiations. Noncycling (confluent) cells of BHK resulted in a shouldered survival curve following gamma irradiation while a biphasic curve was

M. A. Hannan; D. P. Gibson



Anthropometric Characteristics, Physical Activity, and Risk of Non-Hodgkin's Lymphoma Subtypes and B-Cell Chronic Lymphocytic Leukemia: A Prospective Study  

Microsoft Academic Search

Anthropometric characteristics, physical activity, and risk of non-Hodgkin's lymphoma, its subtypes, and B-cell chronic lymphocytic leukemia (B-CLL) were evaluated in a prospective cohort study of 37,931 Iowa women who were aged 55-69 years at baseline in 1986. Through 1998, 261 cases of non-Hodgkin's lymphoma (137 diffuse, 58 follicular, and 32 small lymphocytic lymphomas) and 63 cases of B-CLL were identified

James R. Cerhan; Carol A. Janney; Celine M. Vachon; Thomas M. Habermann; Neil E. Kay; John D. Potter; Thomas A. Sellers; Aaron R. Folsom


CD69 is independently prognostic in chronic lymphocytic leukemia: a comprehensive clinical and biological profiling study  

PubMed Central

Background CD69 is expressed in several hemopoietic cells and is an early activation marker in chronic lymphocytic leukemia. Chronic lymphocytic leukemia is a clinically heterogeneous disease which needs novel prognostic parameters which can be easily and efficiently managed. Design and Methods We investigated CD69 by flow cytometry in a series of 417 patients affected by chronic lymphocytic leukemia and compared this to other biological and clinical prognosticators. Results CD69 was associated with Rai stages (P=0.00002), ?2-microglobulin (P=0.0005) and soluble CD23 (P<0.0001). CD69 and ZAP-70 (P=0.018) or CD38 (P=0.00015) or immunoglobulin variable heavy chain gene mutations (P=0.0005) were also significantly correlated. Clinically, CD69 positive chronic lymphocytic leukemias received chemotherapy more frequently (74%; P<0.0001), and presented a shorter duration of response after fludarabine plus rituximab (P=0.010) as well as shorter progression free survival and overall survival (P<0.0001). CD69 demonstrated true additive prognostic properties, since the CD69+ plus ZAP-70+ or CD38+ or immunoglobulin variable heavy chain gene unmutated patients had the worst progression free survival and overall survival (P<0.0001). Interestingly, low CD69 expression was necessary to correctly prognosticate the longer progression free survival of patients with a low tumor burden of ?2-microglobulin (P=0.002), of soluble CD23 (P=0.020), or of Rai stages 0-I (P=0.005). CD69 was confirmed to be an independent prognostic factor in multivariate analysis of progression free survival (P=0.017) and overall survival (P=0.039). Conclusions Our data indicate that CD69 is significantly correlated with poor clinical and biological prognostic factors and is confirmed to be an independent disease prognosticator. This supports its introduction in a routine laboratory assessment and, possibly, in a prognostic scoring system for chronic lymphocytic leukemia, after an adequate standardization process.

Del Poeta, Giovanni; Del Principe, Maria Ilaria; Zucchetto, Antonella; Luciano, Fabrizio; Buccisano, Francesco; Maria Rossi, Francesca; Bruno, Antonio; Biagi, Annalisa; Bulian, Pietro; Maurillo, Luca; Neri, Benedetta; Bomben, Riccardo; Simotti, Cristina; Coletta, Angela Maria; Dal Bo, Michele; de Fabritiis, Paolo; Venditti, Adriano; Gattei, Valter; Amadori, Sergio



In vitro studies of lymphocytes from patients with plasma cell myeloma. II. Characterization by cell surface markers  

PubMed Central

Highly purified blood lymphocytes from treated and untreated patients with plasmacell myeloma were tested for the presence of *EAC lymphocytes carrying the C3 receptor surface marker using a rosette-technique and for thymus-derived (T) lymphocytes as judged by spontaneous rosette formation with sheep red blood cells. The treated patients were tested 5 weeks after a standardized 4-day treatment with melphalan and prednisolone. Untreated patients had 40–65% EAC-lymphocytes in peripheral blood while the EAC-cell level in treated patients was < 20%. A non-myeloma control group had 20–40%. The percentage of EAC-lymphocytes was correlated with different in vitro functional tests: lymphocyte-mediated lysis of target cells (Chang cell line or chicken red blood cells) treated with rabbit antibody correlated with the percentage of EAC-lymphocytes. DNA synthesis induced in lymphocytes by pokeweed mitogen (PWM) showed a biphasic dose response curve which may indicate activition of two populations of lymphocytes. A correlation was found between EAC-lymphocytes and DNA synthesis induced by a low concentration of PWM (0·1 ?g/ml). The results support observations by other investigators which suggest that antibody-induced cytotoxicity requires cells in the non-T-lymphocyte population and the assumption that PWM is in part a mitogen for human bone marrow-derived lymphocytes.

Mellstedt, H.; Jondal, M.; Holm, G.



Association of Blood Monocyte and Platelet Markers with Carotid Artery Characteristics: The Atherosclerosis Risk in Communities Carotid MRI Study  

Microsoft Academic Search

Background: Atherosclerosis is characterized by infiltration of inflammatory cells from circulating blood. Blood cell activation could play an important role in plaque formation. Methods: We analyzed the relationship between blood cellular markers and quantitative measures of carotid wall components in 1,546 participants from the ARIC (Atherosclerosis Risk in Communities) Carotid MRI Study. Carotid imaging was performed using a gadolinium contrast-enhanced

N. Matijevic; K. K. Wu; A. G. Howard; B. Wasserman; W. Y.-W. Wang; A. R. Folsom; A. R. Sharrett



In vitro T-cell activation of monocyte-derived macrophages by soluble messengers or cell-to-cell contact in bovine tuberculosis  

PubMed Central

The macrophage plays a dual role in tuberculosis, promoting not only protection against mycobacteria, but also survival of the pathogen. Macrophages inhibit multiplication of mycobacteria but also act in concert with lymphocytes through presentation of antigens to T cells. Studies in animal and human infections have suggested a correlation of in vitro growth rates of mycobacteria with in vivo virulence, using uracil uptake to assess mycobacterial metabolism. This study found that blood-derived, non-activated bovine macrophages were capable of controlling Mycobacterium bovis bacillus Calmette–Guérin growth for up to 96 hr, but were permissive to intracellular growth of virulent M. bovis. The present investigation compared the in vitro modulation of these macrophage activities by cytokine-rich T-cell supernatants or cell-to-cell contact. On the one hand, treatment of cultured monocytes with mitogen-produced T-cell supernatants promoted morphological changes suggestive of an activation status, enhanced the antigen presentation capabilities of monocytes and up-regulated major histocompatibility complex class II expression. However, this activation was not associated with enhanced anti-M. bovis activity. On the other hand, incubation of infected monocytes with T-cell populations resulted in proportionally increased inhibition of M. bovis uracil uptake. This inhibition was also seen using cells from uninfected animals and indicated the necessity for cell-to-cell contact to promote antimycobacterial capability.

Liebana, E; Aranaz, A; Welsh, M; Neill, S D; Pollock, J M



Monocyte Activation in Peripheral Blood and Dialyser Eluates: Phenotypic Profile and Cytokine Release  

Microsoft Academic Search

Background\\/Aims: Monocyte activation and subsequent cytokine generation is presumed to be involved in haemodialysis (HD)-related morbidity. The present study was designed to investigate HD-induced changes in monocytes, with respect to their phenotypic profile and cytokine release, both in peripheral blood (PB) and dialyser eluates (DE). In addition, the effect of the type of dialyser on monocyte activation was assessed. Methods:

W. E. M. Schouten; M. P. C. Grooteman; M. Schoorl; A. J. van Houte; M. J. Nubé



Inflammatory status of transmigrating primary rat monocytes in a novel perfusion model simulating blood flow  

PubMed Central

It remains unclear whether monocyte infiltration plays a protective or detrimental role in neurodegenerative disease. The present study characterizes the inflammatory status of primary monocytes in a novel in vitro perfusion model. Monocytes under perfusion do not undergo elevated cell death. However, perfusion does lead to altered morphology, which can be counteracted by anti-inflammatory drugs. Functional studies indicate that cytokine levels are significantly reduced in perfusion compared to stationary conditions and enhanced with brain slices or capillary endothelial cells. Understanding monocyte properties could lead to refined treatment and new ways to interfere with inflammation in diseased brains.

Hohsfield, Lindsay A.; Ammann, Christoph G.; Humpel, Christian



Inflammatory status of transmigrating primary rat monocytes in a novel perfusion model simulating blood flow.  


It remains unclear whether monocyte infiltration plays a protective or detrimental role in neurodegenerative disease. The present study characterizes the inflammatory status of primary monocytes in a novel in vitro perfusion model. Monocytes under perfusion do not undergo elevated cell death. However, perfusion does lead to altered morphology, which can be counteracted by anti-inflammatory drugs. Functional studies indicate that cytokine levels are significantly reduced in perfusion compared to stationary conditions and enhanced with brain slices or capillary endothelial cells. Understanding monocyte properties could lead to refined treatment and new ways to interfere with inflammation in diseased brains. PMID:23499257

Hohsfield, Lindsay A; Ammann, Christoph G; Humpel, Christian



Serial study of the effect of radiotherapy on semen parameters, hamster egg penetration rates, and lymphocyte chromosome abnormalities  

SciTech Connect

This study was designed to assess the long-term effects of radiotherapy (RT) on male fertility and the induction of lymphocyte and sperm chromosome abnormalities. This preliminary report provides information on 11 cancer patients (mainly seminomas) treated by RT (testicular dose, 44 to 499 rads). All 11 men were studied pre-RT and at intervals post-RT. The pre-RT semen profile varied considerably, but, in general, the profile was poor with a mean sperm concentration of 19.4 x 10/sup 6/ ml and a mean hamster egg penetration rate of 5%. One month after RT, the sperm concentration decreased and hamster egg penetration was 0% in all men. At 3 and 12 months post-RT, all but two patients were azoospermic. By 24 months post-RT, 9 of 11 patients had regained sperm production and 5 had sperm capable of hamster egg penetration. The three men who have been studied 36 months post-RT had a mean sperm concentration of 45.3 x 10/sup 6/ ml, and all had positive hamster egg penetration tests, although two of the three men had very low penetration rates (2% and 4%). Lymphocyte chromosome analysis demonstrated a striking frequency of chromosome abnormalities post-RT which decreased with time (pre-RT, 0%; 1 month, 42.4%; 3 months, 24.7%; 12 months, 13.8%; 24 months, 11.2%; and 36 months, 10.0%). Thus, it appears that sperm production starts to recover 2 to 3 years after RT when the frequency of lymphocyte chromosome abnormalities has decreased, but the sperm may not be fully functional at this time, as evidenced by poor rates of hamster egg penetration. Future studies of sperm chromosome analysis in these men will determine whether this impairment of the sperm is associated with meiotic chromosome abnormalities.

Martin, R.H.; Barnes, M.; Arthur, K.; Ringrose, T.; Douglas, G.



In vivo imaging implicates CCR2+ monocytes as regulators of neutrophil recruitment during arthritis  

PubMed Central

The infiltration of neutrophils and monocytes is a prominent feature of inflammatory diseases including human rheumatoid arthritis. Understanding how neutrophil recruitment is regulated during pathogene sis is crucial for developing anti-inflammatory therapies. We optimized the K/B × N serum-induced mouse arthritis model to study neutrophil trafficking dynamics in vivo using two-photon microscopy. Arthritogenic serum was injected subcutaneously into one hind footpad to induce a local arthritis with robust neutrophil recruitment. Using this approach, we showed that the depletion of monocytes with clodronate liposomes impaired neutrophil recruitment specifically at the transendothelial migration step. The depletion of CCR2+ monocytes with the monoclonal antibody MC-21 reproduced these effects, implicating CCR2+ monocytes as key regulators of neutrophil extravasation during arthritis initiation. However, monocyte depletion did not prevent neutrophil extravasation in response to bacterial challenge. These findings suggest that anti-inflammatory therapies targeting monocytes may act in part through antagonizing neutrophil extravasation at sites of aseptic inflammation.

Wang, Baomei; Zinselmeyer, Bernd H.; Runnels, Herbert A.; LaBranche, Timothy P.; Morton, Phillip A.; Kreisel, Daniel; Mack, Matthias; Nickerson-Nutter, Cheryl; Allen, Paul M.; Miller, Mark J.



NHE-1 and ?1 integrin-dependent monocyte adhesion and migration after glucose, insulin or PPAR? stimulation  

PubMed Central

In the present study the effect of high glucose concentrations, insulin, PPAR? activators (rosiglitazone) and NHE-1 inhibitors (cariporide) in atherosclerosis-related functions of human monocytes was investigated. Monocyte adhesion to laminin-1, collagen type IV and endothelial cells, as well as monocyte migration through the same substrates were studied. Incubation of the monocyte suspension with high glucose concentrations, insulin and rosiglitazone induced all the studied atherosclerosis-related functions of the monocytes. In all these functions the addition of cariporide counteracted the activity of glucose, insulin and rosiglitazone. The use of antigen for ?1 integrin also counteracted the activity of the above in monocyte adhesion in all three substrates. The data of the present study suggests that PPAR? activation in monocytes induces atherosclerosis, and that NHE-1 and ?1 integrin play an important role in the beginning of atherosclerosis.

Zolota, Zacharoula; Paletas, Konstantinos; Kaloyianni, Martha



Human immunodeficiency virus type 1 infection alters enzymatic and ultrastructural features of peripheral blood monocytes  

PubMed Central

INTRODUCTION: Human immunodeficiency virus-1 (HIV-1) infected monocytes are now believed to serve as a reservoir for HIV-1 infection, and to play a role in viral rebound phenomena in certain groups of patients who failed or stopped highly active antiretroviral therapy (HAART). Data characterizing the morphological changes of peripheral blood monocytes in HIV-1-infected individuals are limited. MATERIALS AND METHODS: In this study, we collected monocytes from 21 asymptomatic HIV-1-infected individuals with CD4 count more than 500 cells/mm3 and healthy individuals. The monocytes ultrastructural morphologic changes and ?-naphthyl butyrate esterase (ANBE) activity were compared between the two groups. RESULTS: In monocytes from patients infected with HIV-1, activity of ?-naphthyl butyrate esterase (ANBE) was markedly increased compared with normal monocytes. In both light microscopic and ultrastructural studies, the cytoplasm of monocytes from HIV-1-infected patients contained a haphazard appearing network of thin fibrils. Cell surface expression of the activation marker HLA-DR molecule was upregulated. There were no discernible differences between the cell surface expression of CD4, CD14, and CD16 molecules comparing normal monocytes to those from HIV-1-infected patients. ?-naphthyl butyrate esterase (ANBE) was markedly increased compared with normal monocytes. In both light microscopic and ultrastructural studies, the cytoplasm of monocytes from HIV-1-infected patients contained a haphazard appearing network of thin fibrils. Cell surface expression of the activation marker HLA-DR molecule was upregulated. There were no discernible differences between the cell surface expression of CD4, CD14, and CD16 molecules comparing normal monocytes to those from HIV-1-infected patients. CONCLUSIONS: Possibly, changes in the activity of ANBE along with a disrupted appearing cytoplasmic fibril network contribute to monocyte dysfunction in HIV-1-infected patients.

Gabali, Ali M.; Jazaerly, Tarek; Cleveland, Ronald; Kass, Lawrence



A Miniature Couette to Generate Shear for Flow Cytometry: Studying Real-Time Modulation of Intracellular Calcium in Monocytic Cells  

PubMed Central

Extracellular hydrodynamic forces may be transmitted to the interior of cells through the alteration of integrin conformation and affinity. Integrin activation regulates leukocyte recruitment, cell activation, and transmigration. The cellular and molecular mechanisms for integrin activation are not precisely known, although intracellular calcium signaling is involved. Flow cytometry offers a versatile way to study intracellular calcium signaling in real-time. We report a novel method to generate defined shear by using a miniature Couette. Testing involved measuring shear induced intracellular calcium signals of human monoblastoid U937 cells in suspension. The Couette was connected externally to a flow cytometer and pressurized at 6 PSI (4.1 N/m2). Cells were subjected to well-defined shear between 0 and 1000 s?1 and delivered continuously within 10 s to a FACScan at 1 ?l/s. Intracellular calcium levels and the percentage of cells activated increased as shear increased in duration and intensity.

Zwartz, Gordon J.; Chigaev, Alexandre; Foutz, Terry D.; Edwards, Bruce; Sklar, Larry A.



Genotoxicity of inhalation anesthetics halothane and isoflurane in human lymphocytes studied in vitro using the comet assay  

Microsoft Academic Search

The alkaline single cell gel electrophoresis (comet) assay was applied to study genotoxic properties of two inhalation anesthetics—halothane and isoflurane—in human peripheral blood lymphocytes (PBL). The cells were exposed in vitro to either halothane (2-bromo-2-chloro-1,1,1-trifluoroethane) or isoflurane (1-chloro-2,2,2-trifluoroethyl difluoromethyl ether) at concentrations 0.1–10 mM in DMSO. The anesthetics-induced DNA strand breaks as well as alkali-labile sites were measured as total

Pawe? Ja?oszy?ski; Maciej Kujawski; Marcin W?sowicz; Roman Szulc; Krzysztof Szyfter



Identification of specific recognition molecules on murine mononuclear phagocytes and B lymphocytes for Vi capsular polysaccharide: modulation of MHC class II expression on stimulation with the polysaccharide.  

PubMed Central

Vi bacterial polysaccharide is a homopolymer of alpha 1-4 N-acetyl polygalacturonic acid with variable O-acetylation at position C-3 and forms a capsule around many bacteria. It has been referred to as the virulence factor of Salmonella typhi and is also a candidate vaccine against typhoid fever. The present study reports the interaction of this polysaccharide with murine mononuclear phagocytes and lymphocytes, and with human monocytes. Vi showed a dose-dependent binding to the murine monocyte cell lines WEHI-274.1 and J774. This binding was abrogated if the polysaccharide was deacetylated, suggesting involvement of acetyl groups in this interaction. Vi also bound to the murine B-cell lymphoma line A20, to peritoneal exudate cells and to a lesser degree to spleen cells and thymocytes from BALB/c mice. The polysaccharide also interacted with the human histiocytic lymphoma line U937 but not with the human monocyte cell line THP-1. Stimulation with Vi led to up-regulation of surface major histocompatibility complex (MHC) class II expression on A20 cells. Immunoprecipitation of Vi-bound molecules from cell surface biotinylated A20 and WEHI-274.1 revealed two bands with MW of about 32,000 and 36,000. The study demonstrates that Vi capsular polysaccharide can interact with mononuclear phagocytes and lymphocytes through specific cell surface molecules and modulate MHC class II expression. Images Figure 5

Qadri, A



Upregulation of programmed death-1 on T cells and programmed death ligand-1 on monocytes in septic shock patients  

PubMed Central

Introduction Studies on the role of programmed death-1(PD-1) and its main ligand (PD-L1) during experimental models of sepsis have shown that the PD-1/PD-L1 pathway plays a pathologic role in altering microbial clearance, the innate inflammatory response and accelerated apoptosis in sepsis. However, the expression of PD-1 and PD-L1 and their role during the development of immune suppression in septic patients have not been elucidated. The present study was designed to determine whether the expression of PD-1 and PD-L1 is upregulated in septic shock patients and to explore the role of this pathway in sepsis-induced immunosuppression. Methods Nineteen septic shock patients and 22 sex-matched and age-matched healthy controls were prospectively enrolled. Apoptosis in lymphocyte subpopulations and PD-1/PD-L1 expression on peripheral T cells, B cells and monocytes were measured using flow cytometry. Apoptosis of T cells induced by TNF? or T-cell receptor ligation in vitro and effects of anti-PD-L1 antibody administration were measured by flow cytometry. CD14+ monocytes of septic shock patients were purified and incubated with either lipopolysaccharide, anti-PD-L1 antibody, isotype antibody, or a combination of lipopolysaccharide and anti-PD-L1 antibody or isotype antibody. Supernatants were harvested to examine production of cytokines by ELISA. Results Compared with healthy controls, septic shock induced a marked increase in apoptosis as detected by the annexin-V binding and active caspase-3 on CD4+ T cells, CD8+ T cells and CD19+ B cells. Expression of PD-1 on T cells and of PD-L1 on monocytes was dramatically upregulated in septic shock patients. PD-1/PD-L1 pathway blockade in vitro with anti-PD-L1 antibody decreased apoptosis of T cells induced by TNF? or T-cell receptor ligation. Meanwhile, this blockade potentiated the lipopolysaccharide-induced TNF? and IL-6 production and decreased IL-10 production by monocytes in vitro. Conclusions The expression of PD-1 on T cells and PD-L1 on monocytes was upregulated in septic shock patients. The PD-1/PD-L1 pathway might play an essential role in sepsis-induced immunosuppression.



Lymphocyte count was significantly associated with hyper-LDL cholesterolemia independently of high-sensitivity C-reactive protein in apparently healthy Japanese.  


The aim of this study was to investigate the association between leukocyte subtype counts and hyper-LDL cholesterolemia, hypertriglyceridemia, and hypo-HDL cholesterolemia. Logistic regressions using hyper-LDL cholesterolemia, hypertriglyceridemia, and hypo-HDL cholesterolemia as a dependent variable and total leukocyte, basophil, eosinophil, neutrophil, lymphocyte, and monocyte counts as an independent variable were calculated adjusting for age, body mass index (BMI), high-sensitivity C-reactive protein (hs-CRP), smoking, drinking, and physical activity in apparently healthy Japanese men (1,803) and women (1,150). The odds ratio (OR) of hyper-LDL cholesterolemia for total leukocyte, eosinophil, and lymphocyte counts, the OR of hypertriglyceridemia for total leukocyte, eosinophil, neutrophil, and lymphocyte counts, and the OR of hypo-HDL cholesterolemia for total leukocyte, neutrophil, and lymphocyte counts were significant in men, and the OR of hyper-LDL cholesterolemia, for lymphocyte count, and the OR of hypo-HDL cholesterolemia for eosinophil count were significant in women. Lymphocyte count was significantly associated with hyper-LDL cholesterolemia independently of hs-CRP in apparently healthy Japanese. PMID:21655904

Oda, Eiji; Kawai, Ryu; Aizawa, Yoshifusa



Prediction of clinical toxicity in localized cervical carcinoma by radio-induced apoptosis study in peripheral blood lymphocytes (PBLs)  

PubMed Central

Background Cervical cancer is treated mainly by surgery and radiotherapy. Toxicity due to radiation is a limiting factor for treatment success. Determination of lymphocyte radiosensitivity by radio-induced apoptosis arises as a possible method for predictive test development. The aim of this study was to analyze radio-induced apoptosis of peripheral blood lymphocytes. Methods Ninety four consecutive patients suffering from cervical carcinoma, diagnosed and treated in our institution, and four healthy controls were included in the study. Toxicity was evaluated using the Lent-Soma scale. Peripheral blood lymphocytes were isolated and irradiated at 0, 1, 2 and 8 Gy during 24, 48 and 72 hours. Apoptosis was measured by flow cytometry using annexin V/propidium iodide to determine early and late apoptosis. Lymphocytes were marked with CD45 APC-conjugated monoclonal antibody. Results Radiation-induced apoptosis (RIA) increased with radiation dose and time of incubation. Data strongly fitted to a semi logarithmic model as follows: RIA = ?ln(Gy) + ?. This mathematical model was defined by two constants: ?, is the origin of the curve in the Y axis and determines the percentage of spontaneous cell death and ?, is the slope of the curve and determines the percentage of cell death induced at a determined radiation dose (? = ?RIA/?ln(Gy)). Higher ? values (increased rate of RIA at given radiation doses) were observed in patients with low sexual toxicity (Exp(B) = 0.83, C.I. 95% (0.73-0.95), p = 0.007; Exp(B) = 0.88, C.I. 95% (0.82-0.94), p = 0.001; Exp(B) = 0.93, C.I. 95% (0.88-0.99), p = 0.026 for 24, 48 and 72 hours respectively). This relation was also found with rectal (Exp(B) = 0.89, C.I. 95% (0.81-0.98), p = 0.026; Exp(B) = 0.95, C.I. 95% (0.91-0.98), p = 0.013 for 48 and 72 hours respectively) and urinary (Exp(B) = 0.83, C.I. 95% (0.71-0.97), p = 0.021 for 24 hours) toxicity. Conclusion Radiation induced apoptosis at different time points and radiation doses fitted to a semi logarithmic model defined by a mathematical equation that gives an individual value of radiosensitivity and could predict late toxicity due to radiotherapy. Other prospective studies with higher number of patients are needed to validate these results.



The effects of prednisone and azathioprine on circulating immunoglobulin levels and lymphocyte subpopulations in normal dogs.  

PubMed Central

This study investigates serum immunoglobulin (SIg) levels and lymphocyte subpopulations in normal dogs in response to putative immunosuppressive doses of prednisone and/or azathioprine. The objectives were to quantify SIg levels and lymphocyte subpopulations, including Thy-1+, CD4+, CD8+ and B cells, in normal dogs both before and after the administration of prednisone and/or azathioprine at 2 mg/kg, PO, each. Eighteen beagles were divided into 3 groups of 6 dogs each. Blood samples for radial immunodiffusion assay of IgG, IgM and IgA, complete blood count (CBC)and flow cytometry were collected prior to the administration of any drugs and again after 14 d of azathioprine, prednisone or azathioprine and prednisone. Peripheral blood mononuclear cells were isolated using density centrifugation and were incubated with monoclonal antibodies reacting with CD4+, CD8+, Thy-1+ and membrane immunoglobulin. Lymphocyte subsets were quantified using flow cytometry. Azathioprine-treated dogs had no significant changes in SIg levels or lymphocyte subpopulations. Prednisone-treated dogs had significant (P < 0.05) decreases in all SIg levels, all lymphocyte subpopulations and erythrocyte numbers, and had an increase in neutrophil counts. Prednisone and azathioprine-treated dogs had significant (P < 0.05) decreases in serum IgG levels and Thy-1+ and CD8+ lymphocyte subpopulations, with an increase in the CD4:CD8. These dogs also had a significant decrease in erythrocyte number and a significant increase in the monocyte count. These findings suggest that azathioprine and prednisone in combination or prednisone alone may be useful for the treatment of T cell-mediated diseases since decreased circulating T cell levels were demonstrated following treatment. The combination of drugs or azathioprine alone may not be appropriate for treatment of acute or autoantibody-mediated immune disease, because SIg levels were minimally affected by treatment.

Rinkardt, N E; Kruth, S A; Kaushik, A



[Biological effects of mineral fibers on lymphocytes in vitro. Comparative studies of asbestos and glass fibers].  


The biological effects of asbestos and glass fibers on lymphocytes in vitro were investigated. Blastoid transformation and beta 2 microglobulin production of human peripheral blood lymphocytes (PBL) induced by phytohemagglutinin were inhibited by Canadian chrysotile B (Standard sample of the Union Internationale Contre le Cancer; UICC standard sample) but it was not the case for crocidolite (UICC standard sample) and small and large glass fibers (John Manville, Canada). Cytotoxic activities of natural killer and killer cells of PBL were investigated using K562 cells and Chang liver cells as target cells respectively. Chrysotile inhibited the both activities but crocidolite and two kind of glass fibers not. In regard to release of lactic dehydrogenase and beta-glucuronidase from mouse peritoneal macrophages exposed to mineral fibers, chrysotile showed more effective reaction compared to crocidolite and amosite (UICC standard sample), on the other hand large glass fiber showed the similar reaction to chrysotile but small glass fiber and milled amosite did not induce any more enzymatic release than the control. In addition to chemical compositions of mineral fibers, the morphological characteristics were also discussed in relation to their biological effects. PMID:6366291

Nakatani, Y



Changes in monocytic expression of aminopeptidase N/CD13 after major trauma  

PubMed Central

HLA-DR expression on monocytes as marker for monocytic function is severely depressed after major trauma. The membrane enzyme aminopeptidase N/CD13 can trigger help in antigen processing by MHC class II molecules of antigen-presenting cells. We determined the simultaneous expression of HLA-DR and CD13 on peripheral blood monocytes of patients with major trauma (injury severity score of ?16). 1 : 1 conjugates of phycoerythrin (PE)-to-monoclonal antibody were used in combination with QuantiBRITETM PE beads for a standardized quantification in terms of antibodies bound per cell (ABC). The very low expression of HLA-DR antigen on monocytes of patients at day 1 after major trauma confirmed previous results in the literature. Monocytic HLA-DR expression increased slowly to reach values in the lower range of healthy volunteers at day 14. Monocytic CD13 expression at day 1 showed values in the range of healthy volunteers, and a strong rise afterwards. Fourteen days after trauma, the monocytic expression of CD13 was still much higher than in the control group. Because lipopolysaccharide (LPS) and the anti-inflammatory cytokine interleukin (IL)-10 have been shown to be involved in the depressed HLA-DR expression on monocytes in trauma patients, we studied the in vitro effects of LPS and interleukin (IL)-10 on the expression of CD13 on monocytes prepared from the peripheral blood of healthy volunteers. Whereas a 3-day IL-10 treatment resulted in a down-regulation of both HLA-DR and CD13 expression on monocytes, LPS caused a down-regulation of HLA-DR but a rapid up-regulation of CD13 levels. Therefore we suggest that, with respect to monocytic CD13 expression, LPS rather than IL-10 could well be the explanation for monocytic surface molecules after severe injury, although other mediators with a CD13 regulating function have to be considered.




Carcinoma origin dictates differential skewing of monocyte function  

PubMed Central

Macrophages are versatile cells, which phenotype is profoundly influenced by their environment. Pro-inflammatory classically activated or M1 macrophages, and anti-inflammatory alternatively-activated or M2 macrophages represent two extremes of a continuum of functional states. Consequently, macrophages that are present in tumors can exert tumor-promoting and tumor-suppressing activity, depending on the tumor milieu. In this study we investigated how human monocytes—the precursors of macrophages—are influenced by carcinoma cells of different origin. We demonstrate that monocytes, stimulated with breast cancer supernatant, showed increased expression of interleukin (IL)-10, IL-8 and chemokines CCL17 and CCL22, which are associated with an alternatively-activated phenotype. By contrast, monocytes that were cultured in supernatants of colon cancer cells produced more pro-inflammatory cytokines (e.g., IL-12 and TNF?) and reactive oxygen species. Secretome analysis revealed differential secretion of proteins by colon and breast cancer cell lines, of which the proteoglycan versican was exclusively secreted by colon carcinoma cell lines. Reducing active versican by blocking with monoclonal antibodies or shRNA diminished pro-inflammatory cytokine production by monocytes. Thus, colon carcinoma cells polarize monocytes toward a more classically-activated anti-tumorigenic phenotype, whereas breast carcinomas predispose monocytes toward an alternatively activated phenotype. Interestingly, presence of macrophages in breast or colon carcinomas correlates with poor or good prognosis in patients, respectively. The observed discrepancy in macrophage activation by either colon or breast carcinoma cells may therefore explain the dichotomy between patient prognosis and macrophage presence in these different tumors. Designing new therapies, directing development of monocytes toward M1 activated tumor macrophages in cancer patients, may have great clinical benefits.

Bogels, Marijn; Braster, Rens; Nijland, Philip G.; Gul, Nuray; van de Luijtgaarden, Wendy; Fijneman, Remond J.A.; Meijer, Gerrit A.; Jimenez, Connie R.; Beelen, Robert H.J.; van Egmond, Marjolein



Hypoxia-inducible C-to-U coding RNA editing downregulates SDHB in monocytes.  


Background. RNA editing is a post-transcriptional regulatory mechanism that can alter the coding sequences of certain genes in response to physiological demands. We previously identified C-to-U RNA editing (C136U, R46X) which inactivates a small fraction of succinate dehydrogenase (SDH; mitochondrial complex II) subunit B gene (SDHB) mRNAs in normal steady-state peripheral blood mononuclear cells (PBMCs). SDH is a heterotetrameric tumor suppressor complex which when mutated causes paraganglioma tumors that are characterized by constitutive activation of hypoxia inducible pathways. Here, we studied regulation, extent and cell type origin of SDHB RNA editing. Methods. We used short-term cultured PBMCs obtained from random healthy platelet donors, performed monocyte enrichment by cold aggregation, employed a novel allele-specific quantitative PCR method, flow cytometry, immunologic cell separation, gene expression microarray, database analysis and high-throughput RNA sequencing. Results. While the editing rate is low in uncultured monocyte-enriched PBMCs (average rate 2.0%, range 0.4%-6.3%, n = 42), it is markedly upregulated upon exposure to 1% oxygen tension (average rate 18.2%, range 2.8%-49.4%, n = 14) and during normoxic macrophage differentiation in the presence of serum (average rate 10.1%, range 2.7%-18.8%, n = 17). The normoxic induction of SDHB RNA editing was associated with the development of dense adherent aggregates of monocytes in culture. CD14-positive monocyte isolation increased the percentages of C136U transcripts by 1.25-fold in normoxic cultures (n = 5) and 1.68-fold in hypoxic cultures (n = 4). CD14-negative lymphocytes showed no evidence of SDHB editing. The SDHB genomic DNA remained wild-type during increased RNA editing. Microarray analysis showed expression changes in wound healing and immune response pathway genes as the editing rate increased in normoxic cultures. High-throughput sequencing of SDHB and SDHD transcripts confirmed the induction of C136U RNA editing in normoxic cultures but showed no additional verifiable coding edits. Analysis of SDHB RNA sequence data from 16 normal human tissues from the Illumina Body Map and from 45 samples representing 23 different cell types from the ENCODE projects confirmed the occurrence of site-specific C136U editing in whole blood (1.7%) and two primary CD14+ monocyte samples (1.9% and 2.6%). In contrast, the other cell types showed an average of 0.2% and 0.1% C136U editing rates in the two databases, respectively. Conclusions. These findings demonstrate that C-to-U coding RNA editing of certain genes is dynamically induced by physiologically relevant environmental factors and suggest that epigenetic downregulation of SDHB by site-specific RNA editing plays a role in hypoxia adaptation in monocytes. PMID:24058882

Baysal, Bora E; De Jong, Kitty; Liu, Biao; Wang, Jianmin; Patnaik, Santosh K; Wallace, Paul K; Taggart, Robert T



Hypoxia-inducible C-to-U coding RNA editing downregulates SDHB in monocytes  

PubMed Central

Background. RNA editing is a post-transcriptional regulatory mechanism that can alter the coding sequences of certain genes in response to physiological demands. We previously identified C-to-U RNA editing (C136U, R46X) which inactivates a small fraction of succinate dehydrogenase (SDH; mitochondrial complex II) subunit B gene (SDHB) mRNAs in normal steady-state peripheral blood mononuclear cells (PBMCs). SDH is a heterotetrameric tumor suppressor complex which when mutated causes paraganglioma tumors that are characterized by constitutive activation of hypoxia inducible pathways. Here, we studied regulation, extent and cell type origin of SDHB RNA editing. Methods. We used short-term cultured PBMCs obtained from random healthy platelet donors, performed monocyte enrichment by cold aggregation, employed a novel allele-specific quantitative PCR method, flow cytometry, immunologic cell separation, gene expression microarray, database analysis and high-throughput RNA sequencing. Results. While the editing rate is low in uncultured monocyte-enriched PBMCs (average rate 2.0%, range 0.4%–6.3%, n = 42), it is markedly upregulated upon exposure to 1% oxygen tension (average rate 18.2%, range 2.8%–49.4%, n = 14) and during normoxic macrophage differentiation in the presence of serum (average rate 10.1%, range 2.7%–18.8%, n = 17). The normoxic induction of SDHB RNA editing was associated with the development of dense adherent aggregates of monocytes in culture. CD14-positive monocyte isolation increased the percentages of C136U transcripts by 1.25-fold in normoxic cultures (n = 5) and 1.68-fold in hypoxic cultures (n = 4). CD14-negative lymphocytes showed no evidence of SDHB editing. The SDHB genomic DNA remained wild-type during increased RNA editing. Microarray analysis showed expression changes in wound healing and immune response pathway genes as the editing rate increased in normoxic cultures. High-throughput sequencing of SDHB and SDHD transcripts confirmed the induction of C136U RNA editing in normoxic cultures but showed no additional verifiable coding edits. Analysis of SDHB RNA sequence data from 16 normal human tissues from the Illumina Body Map and from 45 samples representing 23 different cell types from the ENCODE projects confirmed the occurrence of site-specific C136U editing in whole blood (1.7%) and two primary CD14+ monocyte samples (1.9% and 2.6%). In contrast, the other cell types showed an average of 0.2% and 0.1% C136U editing rates in the two databases, respectively. Conclusions. These findings demonstrate that C-to-U coding RNA editing of certain genes is dynamically induced by physiologically relevant environmental factors and suggest that epigenetic downregulation of SDHB by site-specific RNA editing plays a role in hypoxia adaptation in monocytes.

De Jong, Kitty; Liu, Biao; Wang, Jianmin; Patnaik, Santosh K.; Wallace, Paul K.; Taggart, Robert T.



Inhibitor of inflammation, peptide fragment (65–76) of monocyte chemotactic protein-1 (MCP-1), inhibits binding of MCP-1 to heparin  

Microsoft Academic Search

Monocyte chemotactic protein-1 (MCP-1, CCL2) is one of the most important chemokines involved in inflammation. MCP-1 stimulates\\u000a migration of monocytes and certain lymphocyte populations to the affected area, in particular to the sites of atherosclerotic\\u000a plaque formation. Development of drugs inhibiting MCP-1 is now a topical task. We earlier designed and synthesized a dodecapeptide\\u000a from C-terminal domain of MCP-1 (65–76,

T. L. Krasnikova; P. I. Nikitin; T. I. Ksenevich; S. G. Gorshkov; T. L. Bushueva; T. I. Arefieva; N. Yu. Ruleva; M. V. Sidorova; A. A. Azmuko; Zh. D. Bespalova



Magnetic resonance imaging of monocyte infiltration in an animal model of multiple sclerosis  

Microsoft Academic Search

The aim of this research was to study neuroinflammatory processes and more specifically the infiltration of monocytes in the central nervous system (CNS) in an animal model of multiple sclerosis (MS) using magnetic resonance imaging (MRI). Monocytes play a key role in MS pathology, and the study of their infiltration kinetics in vivo would benefit from a non-invasive imaging strategy.

R. D. Oude Engberink



Attractin, a dipeptidyl peptidase IV/CD26-like enzyme, is expressed on human peripheral blood monocytes and potentially influences monocyte function.  


The ectoenzyme dipeptidyl peptidase IV (DP IV; CD26) was shown to play a crucial role in T cell activation. Several compounds inhibiting DP IV-like activity are currently under investigation for the treatment of Type 2 diabetes, rheumatoid arthritis, colitis ulcerosa, psoriasis, multiple sclerosis, and other diseases. In the present study, we show that human peripheral blood monocytes express a DP IV-like enzyme activity, which could be inhibited completely by the synthetic DP IV inhibitor Lys[Z(NO(2))]-thiazolidide. DP IV immunoreactivity was not detectable on monocytes, and DP IV transcript levels of monocytes were near the detection limit of quantitative polymerase chain reaction. However, monocytes exhibit a strong mRNA expression of the multifunctional DP IV-like ectoenzyme attractin and were highly positive for attractin in flow cytometric analysis. Fluorescence microscopy clearly demonstrated that attractin is located on the cell surface of monocytes. Attractin immunoprecipitates hydrolyzed Gly-Pro-pNA, indicating that monocyte-expressed attractin possesses DP IV-like activity. Inhibitor kinetic studies with purified human plasma attractin revealed that Lys[Z(NO(2))]-thiazolidide not only inhibits DP IV but also attractin (50% inhibition concentration=8.45 x 10(-9) M). Studying the influence of this inhibitor on monocyte functions, we observed a clear reduction of cell adhesion to fibronectin-coated culture plates in the presence of Lys[Z(NO(2))]-thiazolidide. Moreover, this inhibitor significantly modulates the production of interleukin-1 (IL-1) receptor antagonist, IL-6, and transforming growth factor-beta1 in lipopolysaccharide-stimulated monocyte cultures. In summary, here, we demonstrate for the first time expression of attractin on monocytes and provide first data suggesting that drugs directed to DP IV-like enzyme activity could affect monocyte function via attractin inhibition. PMID:16835316

Wrenger, Sabine; Faust, Jürgen; Friedrich, Daniel; Hoffmann, Torsten; Hartig, Roland; Lendeckel, Uwe; Kähne, Thilo; Thielitz, Anja; Neubert, Klaus; Reinhold, Dirk



Hydrodynamic Regulation of Monocyte Inflammatory Response to an Intracellular Pathogen  

PubMed Central

Systemic bacterial infections elicit inflammatory response that promotes acute or chronic complications such as sepsis, arthritis or atherosclerosis. Of interest, cells in circulation experience hydrodynamic shear forces, which have been shown to be a potent regulator of cellular function in the vasculature and play an important role in maintaining tissue homeostasis. In this study, we have examined the effect of shear forces due to blood flow in modulating the inflammatory response of cells to infection. Using an in vitro model, we analyzed the effects of physiological levels of shear stress on the inflammatory response of monocytes infected with chlamydia, an intracellular pathogen which causes bronchitis and is implicated in the development of atherosclerosis. We found that chlamydial infection alters the morphology of monocytes and trigger the release of pro-inflammatory cytokines TNF-?, IL-8, IL-1? and IL-6. We also found that the exposure of chlamydia-infected monocytes to short durations of arterial shear stress significantly enhances the secretion of cytokines in a time-dependent manner and the expression of surface adhesion molecule ICAM-1. As a functional consequence, infection and shear stress increased monocyte adhesion to endothelial cells under flow and in the activation and aggregation of platelets. Overall, our study demonstrates that shear stress enhances the inflammatory response of monocytes to infection, suggesting that mechanical forces may contribute to disease pathophysiology. These results provide a novel perspective on our understanding of systemic infection and inflammation.

Evani, Shankar J.; Murthy, Ashlesh K.; Mareedu, Naresh; Montgomery, Robbie K.; Arulanandam, Bernard P.; Ramasubramanian, Anand K.



Predictors of Change in CD4 Lymphocyte Count and Weight among HIV Infected Patients on Anti-Retroviral Treatment in Ethiopia: A Retrospective Longitudinal Study  

PubMed Central

Background Antiretroviral treatment (ART) has been introduced in Ethiopia a decade ago and continues to be scaled up. However, there is dearth of literature on the impact of ART on changes in CD4 lymphocyte count and weight among patients on treatment. Objective To determine the predictors of change in CD4 lymphocyte count and weight among HIV/AIDS infected patients taking antiretroviral treatment in eastern Ethiopia. Methods A retrospective cohort study was conducted among HIV/AIDS patients taking ART from 2005 to 2010. A sample of 1540 HIV infected adult patients who started antiretroviral therapy in hospitals located in eastern Ethiopia were included in the study. The primary outcomes of interest were changes in CD4 count and weight. Descriptive statistics and multivariable regression analyses were performed to examine the outcomes among the cohort. Results Both the median CD4 lymphocyte counts and weight showed improvements in the follow up periods. The multivariate analysis shows that the duration of ART was an important predictor of improvements in CD4 lymphocyte count (beta 7.91; 95% CI 7.48–8.34; p 0.000) and weight (beta 0.15; 95% CI 0.13–0.18; p 0.000). Advanced WHO clinical stage, lower baseline CD4 cell count, and baseline hemoglobin levels were factors associated with decline in weight. Actively working patients had higher CD4 lymphocyte count and weight compared to those that were ambulatory (p<0.05). Conclusion We detected a substantial increment in weight and CD4 lymphocyte count among the patients who were taking ART in eastern Ethiopia. Patients who are of older age, with low initial CD4 lymphocyte count, late stage of the WHO clinical stages and lower hemoglobin level may need special attention. The reasons for the improved findings on CD4 count and weight throughout the five years of follow up merit further investigation.

Reda, Ayalu A.; Biadgilign, Sibhatu; Deribew, Amare; Gebre, Betemariam; Deribe, Kebede



LILRA2 selectively modulates LPS-mediated cytokine production and inhibits phagocytosis by monocytes.  


The activating immunoglobulin-like receptor, subfamily A, member 2 (LILRA2) is primarily expressed on the surface of cells of the innate immunity including monocytes, macrophages, neutrophils, basophils and eosinophils but not on lymphocytes and NK cells. LILRA2 cross-linking on monocytes induces pro-inflammatory cytokines while inhibiting dendritic cell differentiation and antigen presentation. A similar activating receptor, LILRA4, has been shown to modulate functions of TLR7/9 in dendritic cells. These suggest a selective immune regulatory role for LILRAs during innate immune responses. However, whether LILRA2 has functions distinct from other receptors of the innate immunity including Toll-like receptor (TLR) 4 and Fc?RI remains unknown. Moreover, the effects of LILRA2 on TLR4 and Fc?RI-mediated monocyte functions are not elucidated. Here, we show activation of monocytes via LILRA2 cross-linking selectively increased GM-CSF production but failed to induce IL-12 and MCP-1 production that were strongly up-regulated by LPS, suggesting functions distinct from TLR4. Interestingly, LILRA2 cross-linking on monocytes induced similar amounts of IL-6, IL-8, G-CSF and MIP-1? but lower levels of TNF?, IL-1?, IL-10 and IFN? compared to those stimulated with LPS. Furthermore, cross-linking of LILRA2 on monocytes significantly decreased phagocytosis of IgG-coated micro-beads and serum opsonized Escherichia coli but had limited effect on phagocytosis of non-opsonized bacteria. Simultaneous co-stimulation of monocytes through LILRA2 and LPS or sequential activation of monocytes through LILRA2 followed by LPS led lower levels of TNF?, IL-1? and IL-12 production compared to LPS alone, but had additive effect on levels of IL-10 and IFN? but not on IL-6. Interestingly, LILRA2 cross-linking on monocytes caused significant inhibition of TLR4 mRNA and protein, suggesting LILRA2-mediated suppression of LPS responses might be partly via regulation of this receptor. Taken together, we provide evidence that LILRA2-mediated activation of monocytes is significantly different to LPS and that LILRA2 selectively modulates LPS-mediated monocyte activation and Fc?RI-dependent phagocytosis. PMID:22479404

Lu, Hao K; Mitchell, Ainslie; Endoh, Yasumi; Hampartzoumian, Taline; Huynh, Owen; Borges, Luis; Geczy, Carolyn; Bryant, Katherine; Tedla, Nicodemus



Lack of utility of chimerism studies obtained two to three months after myeloablative haematopoietic cell transplantation for acute lymphocytic leukaemia  

PubMed Central

Lineage specific chimerism studies are commonly obtained at several time points after nonmyeloablative haematopoietic cell transplantation to assess the tempo and degree of engraftment, and to monitor graft rejection. For patients who receive myeloablative transplants, the value of frequent chimerism analyses using sensitive molecular techniques is less certain. In this study, a retrospective analysis was performed to assess the transplant outcome of 89 adult patients with acute lymphocytic leukaemia who had chimerism studies of unfractionated bone marrow cells or peripheral blood subsets performed approximately 80 days after transplantation. These patients received unmanipulated, myeloablative transplants using either HLA-identical or HLA-mismatched, related or unrelated donor stem cells. Incomplete donor engraftment was present only in the CD3+ peripheral blood T-cells in a small percentage of patients. There was no correlation of mixed chimerism with transplant outcome. Routine “Day 80” chimerism studies in this group of patients who receive intensive, myeloablative conditioning regimens are not recommended.

Doney, Kristine C.; Loken, Michael R.; Bryant, Eileen M.; Smith, Anajane G.; Appelbaum, Frederick R.



Human CD14dim Monocytes Patrol and Sense Nucleic Acids and Viruses via TLR7 and TLR8 Receptors  

PubMed Central

Summary Monocytes are effectors of the inflammatory response to microbes. Human CD14+ monocytes specialize in phagocytosis and production of reactive oxygen species and secrete inflammatory cytokines in response to a broad range of microbial cues. Here, we have characterized the functions of human monocytes that lack CD14 (CD14dim) and express CD16. CD14dim monocytes were genetically distinct from natural killer cells. Gene expression analyses indicated similarities with murine patrolling Gr1dim monocytes, and they patrolled the endothelium of blood vessels after adoptive transfer, in a lymphocyte function-associated antigen-1-dependent manner. CD14dim monocytes were weak phagocytes and did not produce ROS or cytokines in response to cell-surface Toll-like receptors. Instead, they selectively produced TNF-?, IL-1?, and CCL3 in response to viruses and immune complexes containing nucleic acids, via a proinflammatory TLR7-TLR 8-MyD88-MEK pathway. Thus, CD14dim cells are bona fide monocytes involved in the innate local surveillance of tissues and the pathogenesis of autoimmune diseases.

Cros, Jerome; Cagnard, Nicolas; Woollard, Kevin; Patey, Natacha; Zhang, Shen-Ying; Senechal, Brigitte; Puel, Anne; Biswas, Subhra K.; Moshous, Despina; Picard, Capucine; Jais, Jean-Philippe; D'Cruz, David; Casanova, Jean-Laurent; Trouillet, Celine; Geissmann, Frederic



Electrophoretic study of immunoglobulins and immunoglobulin sub-units on the surface of human peripheral blood lymphocytes  

PubMed Central

Treatment of human peripheral blood lymphocytes with a polyvalent antiserum to human immunoglobulins causes a reduction in the electrophoretic mobility of the cells. Treatment of the lymphocytes with antisera to ? and ? chains has a similar effect, but antisera to ?, ? and ? chains are only effective in this way when all antisera are mixed together. These findings point to the presence of immunoglobulin molecules on the surface of peripheral blood lymphocytes and the results are discussed in terms of the possible representation of immunoglobulin sub-units on the lymphocyte surface.

Bert, G.; Massaro, A. L.; Di Cossano, D. Lajolo; Maja, M.



Reduction of indoxyl sulfate by AST-120 attenuates monocyte inflammation related to chronic kidney disease.  


Accelerated cardiovascular disease is a frequent complication of CKD. Monocyte-mediated inflammation and adhesion of monocytes to vascular endothelium are key events in atherogenesis. An oral adsorbent, AST-120, retards renal function deterioration by lowering IS, which is known to accumulate in CKD patients. However, the effect of AST-120 on CKD-related monocyte activation is unknown. We aimed to determine whether AST-120 improves monocyte-mediated inflammation through IS reduction. Flow cytometric analysis showed that Mac-1 expression and ROS production were significantly higher in peripheral blood monocytes of subtotal Nx CKD mice than in sham-operated mice. AST-120 treatment significantly decreased Mac-1 expression and ROS production in CKD model mice. Furthermore, administration of IS induced monocyte-mediated inflammation and ROS generation. In vitro studies indicated that IS dose-dependently increased THP-1 monocytic cell adhesion to IL-1?-activated HUVECs under physiological flow conditions. IS also induced monocyte-mediated inflammation and ROS production in THP-1 cells. Phosphorylation of p38 MAPK and membrane translocation of NAD(P)H oxidase subunit p47phox in THP-1 cells were induced by IS. Both SB203580 (p38 MAPK inhibitor) and apocynin [NAD(P)H oxidase inhibitor] reduced THP-1 cell adhesion to HUVECs. Apocynin also inhibited IS-induced ROS production in THP-1 cells. IS induced monocyte-driven inflammation through NAD(P)H oxidase- and p38 MAPK-dependent pathways in monocytes. The main finding of this study was that AST-120 inhibited monocyte activation by reducing IS in vivo. This provides new insights on how AST-120 attenuates the progression of atherosclerosis in CKD. PMID:23362306

Ito, Shunsuke; Higuchi, Yusuke; Yagi, Yoko; Nishijima, Fuyuhiko; Yamato, Hideyuki; Ishii, Hideto; Osaka, Mizuko; Yoshida, Masayuki



Phenotypic correlations between monocytes and CD4+ T cells in allergic patients.  


Despite widely acknowledged contributions of innate and adaptive immune systems to the pathogenesis of allergic diseases, mutual interactions occurring in vivo between components of those two systems have not been studied in sufficient detail. Here, we wished to investigate whether phenotypic features of monocytes and CD4+ T cells in allergic patients are reciprocally related. Therefore, we recruited 50 untreated house dust mite-sensitive allergic rhinitis patients and 29 non-atopic healthy individuals and performed comprehensive simultaneous flow cytometric analysis of mutual correlations between levels of CD14, CD16, CD163, CD206, CD124 (IL-4R), CD210 (IL-10R) and CD25, CD124, CD127 (IL-7R), CD210, ICOS expression on monocytes and CD4+ T cells, respectively. We found that CD163 monocyte expression in allergic but not healthy subjects is positively correlated with monocyte IL-10R, and, to a lesser extent, CD206, but not IL-4R expression. Levels of CD163 expression were not related to frequencies of CD14++CD16-, CD14++CD16+, and CD14+CD16++ monocyte subsets. In contrast to healthy controls, intensities of monocyte IL-10R in allergic individuals were significantly correlated with monocyte CD206 and IL-4R expression. In addition, levels of monocyte IL-4R and IL-10R monocyte expression were positively correlated to expression of IL-4R and IL-10R on CD4+ T cells in both groups of studied subjects. Interestingly, we demonstrated a significant positive correlation between levels of monocyte CD206 expression and levels of IL-10R and IL-4R expression on CD4+ T cells in allergic but not healthy individuals. In summary, we conclude that allergic rhinitis is associated with a number of phenotypic alterations of circulating monocytes and CD4+ T cells. PMID:23343753

Moniuszko, Marcin; Kowal, Krzysztof; Jeznach, Marta; Rusak, Malgorzata; Dabrowska, Milena; Bodzenta-Lukaszyk, Anna



Bacille Calmette-Gu?rin induces NOD2-dependent nonspecific protection from reinfection via epigenetic reprogramming of monocytes  

PubMed Central

Adaptive features of innate immunity, recently described as “trained immunity,” have been documented in plants, invertebrate animals, and mice, but not yet in humans. Here we show that bacille Calmette-Guérin (BCG) vaccination in healthy volunteers led not only to a four- to sevenfold increase in the production of IFN-?, but also to a twofold enhanced release of monocyte-derived cytokines, such as TNF and IL-1?, in response to unrelated bacterial and fungal pathogens. The enhanced function of circulating monocytes persisted for at least 3 mo after vaccination and was accompanied by increased expression of activation markers such as CD11b and Toll-like receptor 4. These training effects were induced through the NOD2 receptor and mediated by increased histone 3 lysine 4 trimethylation. In experimental studies, BCG vaccination induced T- and B-lymphocyte–independent protection of severe combined immunodeficiency SCID mice from disseminated candidiasis (100% survival in BCG-vaccinated mice vs. 30% in control mice). In conclusion, BCG induces trained immunity and nonspecific protection from infections through epigenetic reprogramming of innate immune cells.

Kleinnijenhuis, Johanneke; Quintin, Jessica; Preijers, Frank; Joosten, Leo A. B.; Ifrim, Daniela C.; Saeed, Sadia; Jacobs, Cor; van Loenhout, Joke; de Jong, Dirk; Stunnenberg, Hendrik G.; Xavier, Ramnik J.; van der Meer, Jos W. M.; van Crevel, Reinout; Netea, Mihai G.



Genome-wide promoter analysis of histone modifications in human monocyte-derived antigen presenting cells  

Microsoft Academic Search

BACKGROUND: Monocyte-derived macrophages and dendritic cells (DCs) are important in inflammatory processes and are often used for immunotherapeutic approaches. Blood monocytes can be differentiated into macrophages and DCs, which is accompanied with transcriptional changes in many genes, including chemokines and cell surface markers. RESULTS: To study the chromatin modifications associated with this differentiation, we performed a genome wide analysis of

Liina Tserel; Raivo Kolde; Ana Rebane; Kai Kisand; Hedi Peterson; Jaak Vilo; Pärt Peterson



Effect of interferon- ? and cell differentiation on Puumala virus infection in human monocyte\\/macrophages  

Microsoft Academic Search

Pathogenesis of hantavirus infections is poorly understood. Puumala virus (PUU) is the etiologic agent of nephropathiaepidemica, a form of hemorrhagic fever with renal syndrome common in Europe. We have studied PUU infection in primary human monocyte\\/macrophages and specifically the role of interferon ? (IFN-?) and cell differentiation in it. PUU infection proceeded at a low level in monocyte\\/macrophages, and nucleocapsid

Mari Temonen; Hilkka Lankinen; Olli Vapalahti; Tapani Ronni; Ilkka Julkunen; Antti Vaheri



Monocytes Are Highly Sensitive to Clostridium difficile Toxin A-Induced Apoptotic and Nonapoptotic Cell Death  

Microsoft Academic Search

In this study we investigated the in vitro responses of peripheral blood mononuclear preparations and purified monocytes to Clostridium difficile toxin A. In contrast to the responses of T and B cells, exposure to toxin A led to a rapid loss of monocytes in a time- and dose-dependent fashion (the majority of cells were lost within 24 h of exposure

K. Solomon; J. Webb; N. Ali; R. A. Robins; Y. R. Mahida



Why do cytotoxic T lymphocytes fail to eliminate hepatitis C virus? Lessons from studies using major histocompatibility complex class I peptide tetramers.  

PubMed Central

Hepatitis C virus (HCV) infection is a major public health problem, affecting an estimated 3% of the world's population, and over 10% in some countries. Infection in most cases becomes persistent, and can lead to hepatic inflammation, fibrosis and liver failure. The T lymphocyte reponse, in particular that mediated by cytotoxic T lymphocytes (CTLs), is likely to be involved in determining the outcome of infection, although its overall role is not clear. The use of major histocompatibility complex (MHC) class I peptide tetrameric complexes (tetramers) to study antiviral CTL responses has revolutionized our approach to the study of human infection. We have used a panel of MHC class I tetramers to analyse immune responses in HCV-infected individuals at various stages of disease. We find that the CTL response against HCV is vigorous in its early phases but dwindles over time both in terms of lymphocyte number and function. A number of potential explanations for this 'CTL failure' are discussed.

Lechner, F; Sullivan, J; Spiegel, H; Nixon, D F; Ferrari, B; Davis, A; Borkowsky, B; Pollack, H; Barnes, E; Dusheiko, G; Klenerman, P



Studies on the homing of Mycobacterium-sensitized T lymphocytes to the synovium during passive adjuvant arthritis.  


The migration of intravenously administered adjuvant sensitized T lymphocytes to the knee synovium of recipient rats undergoing passive adjuvant arthritis has been followed. Using fluorescein isothiocyanate (FITC)-labeled adjuvant-sensitized T cells and anticollagen IgG, the present studies demonstrate the presence of fluorescent cells in the inflamed knee synovium of recipient rats undergoing passive arthritis. Proliferation studies indicate that synovial cells from these rats respond to Mycobacterium tuberculosis (MT). Since cross-reactivity between Mycobacterial antigens and cartilage proteoglycans has been previously demonstrated, it is suggested that adjuvant-sensitized T cells that are injected into naive rats migrate to the synovium, proliferate in response to cartilage proteoglycan, and initiate passive arthritis. PMID:2118831

DeJoy, S Q; Ferguson-Chanowitz, K; Oronsky, A L; Zabriskie, J B; Kerwar, S S



Thymosin-?1 modulates dendritic cell differentiation and functional maturation from human peripheral blood CD14+ monocytes  

PubMed Central

Summary Although thymosins have been demonstrated to have immunomodulatory effects, it is still not clear whether they could affect dendritic cells (DCs), the most professional antigen-presenting cells. The objective of this study was to determine the effect and potential mechanisms of thymosin-?1 (T?1) on DC differentiation and functional maturation. Human peripheral blood CD14+ monocytes were purified by using a magnetic separation column and cultured with GM-CSF and IL-4 to differentiate into immature DCs (iDCs). In the presence of T?1, iDC surface markers CD40, CD80, MHC class I and class II molecules were significantly upregulated as measured by flow cytemotry analysis. However, T?4 or T?10 did not show these effects on iDCs. There was approximately a 30% reduction in antigen (FITC-conjugated dextran)-uptake by T?1-treated iDCs as compared with non-T?1-treated iDCs. In addition, T?1-treated matured DCs (mDCs) showed an increased stimulation of allogeneic CD3+ T-cell proliferation as measured by a mixed-lymphocyte reaction assay. T?1-treated mDCs also increased the production of several Th1- and Th2-type cytokines as measured by a Bio-Plex cytokine assay. Furthermore, rapid activation of p38 MAPK and NF?B was seen in T?1-treated iDCs as measured by a Bio-Plex phosphoprotein assay. T?1 significantly enhances on DC differentiation, activation and functions from human peripheral blood CD14+ monocytes possiblly through a mechanism of the activation of p38 MAPK and NF?B pathways. This study provides a basis to further evaluate T?1 as a possible adjuvant for a DC-directed vaccine or therapy.

Yao, Qizhi; Doan, Linh X.; Zhang, Rongxin; Bharadwaj, Uddalak; Li, Min; Chen, Changyi



Lactoferrin inhibits or promotes Legionella pneumophila intracellular multiplication in nonactivated and interferon gamma-activated human monocytes depending upon its degree of iron saturation. Iron-lactoferrin and nonphysiologic iron chelates reverse monocyte activation against Legionella pneumophila.  

PubMed Central

We have been exploring the role of iron in the pathogenesis of the intracellular bacterial pathogen Legionella pneumophila. In previous studies, we have demonstrated that L. pneumophila intracellular multiplication in human monocytes is iron dependent and that IFN gamma-activated monocytes inhibit L. pneumophila intracellular multiplication by limiting the availability of iron. In this study, we have investigated the effect on L. pneumophila intracellular multiplication of lactoferrin, an iron-binding protein which is internalized via specific receptors on monocytes, and of nonphysiologic iron chelates which enter monocytes by a receptor-independent route. Apolactoferrin completely inhibited L. pneumophila multiplication in nonactivated monocytes, and enhanced the capacity of IFN gamma-activated monocytes to inhibit L. pneumophila intracellular multiplication. In contrast, iron-saturated lactoferrin had no effect on the already rapid rate of L. pneumophila multiplication in nonactivated monocytes. Moreover, it reversed the capacity of activated monocytes to inhibit L. pneumophila intracellular multiplication, demonstrating that L. pneumophila can utilize iron from the lactoferrin-lactoferrin receptor pathway. The capacity of iron-lactoferrin to reverse monocyte activation was dependent upon its percent iron saturation and not just its total iron content. Similarly, the nonphysiologic iron chelates ferric nitrilotriacetate and ferric ammonium citrate completely reverse and ferric pyrophosphate partially reversed the capacity of IFN gamma-activated monocytes to inhibit L. pneumophila intracellular multiplication, demonstrating that L. pneumophila can utilize iron derived from nonphysiologic iron chelates internalized by monocytes independently of the transferrin and lactoferrin endocytic pathways. This study suggests that at sites of inflammation, lactoferrin may inhibit or promote L. pneumophila intracellular multiplication in mononuclear phagocytes depending upon its degree of iron saturation. In addition, this study suggests a potential role for PMN in host defense against L. pneumophila--providing apolactoferrin to infected monocytes--and it supports the concept that PMN and monocytes may cooperate in host defense against intracellular parasites and other pathogens.

Byrd, T F; Horwitz, M A



Cytokine Response of Lymphocytes Persistently Infected with Chlamydia pneumoniae  

Microsoft Academic Search

Chlamydia pneumoniae infection of lymphocytes in blood has been documented, and it is apparent that control of this pathogen in lymphocytes as well as immune functions of the infected lymphocytes may be critical in the development of chronic inflammatory diseases associated with infection by this bacterium. Since immune function of lymphocytes infected with C. pneumoniae has not been well studied,

Riho Takano; Hiroyuki Yamaguchi; Shigehiro Sugimoto; Shinichi Nakamura; Herman Friedman; Yoshimasa Yamamoto



Killing of Brucella antigen-sensitized macrophages by T lymphocytes in bovine brucellosis.  


The present study was an investigation into the role of T lymphocytes in the killing of antigen-sensitized macrophages (M Phi) in bovine brucellosis. Following confirmation of bovine T lymphocyte cell lines derived from Brucella abortus Strain 19 vaccinated steers as antigen-specific in proliferation studies using various antigens, we adapted an apoptosis assay for evaluation of cytotoxicity by these bovine T cells against autologous monocyte-derived macrophages (MDM Phi) as target cells. Various B. abortus antigen preparations were tested including whole gamma-irradiated B. abortus bacteria (gamma BA), a soluble cytosolic protein fraction and a membrane-associated protein fraction. Both polyclonal and cloned T lymphocyte cell lines exhibited cytotoxicity against MDM Phi targets in an antigen-specific fashion. Polyclonal and cloned T lymphocyte cell lines demonstrated cytotoxic responses to varying degrees against B. abortus antigens regardless of whether the antigen used was whole nonviable bacteria, a soluble protein extract or a membrane-associated fraction of extracted bacteria. To further develop correlation of these responses to an in vivo host defense mechanism, cytotoxicity was evaluated using target cells that had been infected with live B. abortus S19 or B. abortus Strain 2308. Cytotoxic responses were also demonstrated consistently against infected targets with either strain of B. abortus although in most cases, cytotoxicity was higher against target cells sensitized with gamma BA compared to those infected with live bacteria. Cloned T lymphocyte cell lines were all CD4+, CD8(-) cells indicating that the observed cytotoxic responses were most likely due to an inflammatory Th1 response and may represent an important host defense mechanism induced by vaccination with live attenuated strains of B. abortus in cattle. PMID:17904229

Wyckoff, John H; Potts, Richard D



Use of Single-Cycle Infectious Lymphocytic Choriomeningitis Virus To Study Hemorrhagic Fever Arenaviruses ?  

PubMed Central

Several arenaviruses, chiefly Lassa virus (LASV) and Junin virus in West Africa and Argentina, respectively, cause hemorrhagic fever (HF) disease in humans that is associated with high morbidity and significant mortality. The investigation of antiviral strategies to combat HF arenaviruses is hampered by the requirement of biosafety level 4 (BSL-4) facilities to work with these viruses. These biosafety hurdles could be overcome by the use of recombinant single-cycle infectious arenaviruses. To explore this concept, we have developed a recombinant lymphocytic choriomeningitis virus (LCMV) (rLCMV?GP/GFP) where we replaced the viral glycoprotein (GP) with the green fluorescent protein (GFP). We generated high titers of GP-pseudotyped rLCMV?GP/GFP via genetic trans complementation using stable cell lines that constitutively express LCMV or LASV GPs. Replication of these GP-pseudotyped rLCMV?GP/GFP viruses was restricted to GP-expressing cell lines. This system allowed us to rapidly and reliably characterize and quantify the neutralization activities of serum antibodies against LCMV and LASV within a BSL-2 facility. The sensitivity of the GFP-based microneutralization assay we developed was similar to that obtained with a conventionally used focus reduction neutralization (FRNT) assay. Using GP-pseudotyped rLCMV?GP/GFP, we have also obtained evidence supporting the feasibility of this approach to identify and evaluate candidate antiviral drugs against HF arenaviruses without the need of BSL-4 laboratories.

Rodrigo, W. W. Shanaka I.; de la Torre, Juan C.; Martinez-Sobrido, Luis



Association study between B- and T-lymphocyte attenuator gene and type 1 diabetes mellitus or systemic lupus erythematosus in the Japanese population.  


This study is to elucidate whether the B- and T-lymphocyte attenuator (BTLA) gene is a new susceptibility gene for the development of type 1 diabetes (T1D) and systemic lupus erythematosus (SLE). As a result, this study did not find any genetic contribution of the BTLA gene to the development of T1D and SLE in Japanese population. PMID:19207938

Inuo, M; Ihara, K; Matsuo, T; Kohno, H; Hara, T



A study of 82 extended HLA haplotypes in HFE-C282Y homozygous hemochromatosis subjects: relationship to the genetic control of CD8+ T-lymphocyte numbers and severity of iron overload  

Microsoft Academic Search

BACKGROUND: It has been recently demonstrated that CD8+ T-lymphocyte numbers are genetically transmitted in association with the MHC class I region. The present study was designed with the objective of narrowing the region associated with the setting of CD8+ T-lymphocyte numbers in a population of C282Y homozygous hemochromatosis subjects, in whom a high prevalence of abnormally low CD8+ T-lymphocyte counts

Eugénia Cruz; Jorge Vieira; Susana Almeida; Rosa Lacerda; Andrea Gartner; Carla S Cardoso; Helena Alves; Graça Porto



Conjugated Linoleic Acid Targets ?2 Integrin Expression To Suppress Monocyte Adhesion.  


Chronic recruitment of monocytes and their subsequent migration through the activated endothelium contribute to atherosclerotic plaque development. Integrin-mediated leukocyte adhesion is central to this process. Conjugated linoleic acid (CLA) has the unique property of inducing regression of pre-established murine atherosclerosis via modulation of monocyte/macrophage function. Understanding the mechanisms through which CLA mediates its atheroprotective effect may help to identify novel pathways that limit or reverse atherosclerosis. In this study, we identified a novel mechanism through which CLA alters monocyte function. We show that CLA inhibits human peripheral blood monocyte cell adhesion to activated endothelial cells via loss of CD18 expression, the ?2 chain of LFA-1 and Mac-1 integrins. In addition, using a static-adhesion assay, we provide evidence that CLA prevents monocytes from binding to ICAM-1 and subsequently reduces the capacity of these cells to polarize. CXCL12-CXCR4 interactions induce a conformational change in ?2 integrins, facilitating leukocyte adhesion. In this study, we demonstrate that CLA inhibits CXCR4 expression, resulting in a failure of monocytes to directionally migrate toward CXCL12. Finally, using intravital microscopy, we show that, during CLA-induced regression of pre-established atherosclerosis in ApoE(-/-) mice, there is reduced leukocyte adhesion and decreased CD18 expression on Gr1(+)/CD115(+) proinflammatory monocytes. In summary, the data presented describe a novel functional role for CLA in the regulation of monocyte adhesion, polarization, and migration. PMID:24048900

de Gaetano, Monica; Dempsey, Eugene; Marcone, Simone; James, William G; Belton, Orina



Human lymphocyte repertoires in ageing.  


Deterioration of adaptive immunity with ageing may reflect changes in the repertoire of T cells and B cells available to respond to antigenic challenges, due to altered proportions and absolute numbers of lymphocyte subpopulations as well as changes in the repertoire of antigen receptor genes expressed by these cells. High-throughput DNA sequencing (HTS) now facilitates examination of immunoglobulin and T cell receptor gene rearrangements, and initial studies using these methods to study immune system ageing in humans have demonstrated age-related alterations in the receptor populations within lymphocyte subsets, as well as in repertoires responding to vaccination. Accurate measurement of repertoire diversity remains an experimental challenge. Studies of larger numbers of human subjects, analysis of defined lymphocyte subpopulations including antigen-specific populations, and controlling for factors such as chronic viral infections, will be important for gaining additional understanding of the impact of ageing on human lymphocyte populations. PMID:23992996

Boyd, Scott D; Liu, Yi; Wang, Chen; Martin, Victoria; Dunn-Walters, Deborah K



Caffeine sensitization of cultured mammalian cells and human lymphocytes irradiated with gamma rays and fast neutrons: a study of relative biological effectiveness in relation to cellular repair  

SciTech Connect

The sensitizing effects of caffeine were studied in baby hamster kidney (BHK-21) cells and human lymphocytes following irradiation with gamma rays and fast neutrons. Caffeine sensitization occurred only when log-phase BHK cells and mitogen-stimulated lymphocytes were exposed to the two radiations. Noncycling (confluent) cells of BHK resulted in a shouldered survival curve following gamma irradiation while a biphasic curve was obtained with the log-phase cells. Survival in the case of lymphocytes was estimated by measurement of (TH)thymidine uptake. The relative biological effectiveness (RBE) of fast neutrons was found to be greater at survival levels corresponding to the resistant portions of the survival curves (shoulder or resistant tail). In both cell types, no reduction in RBE was observed when caffeine was present, because caffeine affected both gamma and neutron survival by the same proportion.

Hannan, M.A.; Gibson, D.P.



Monocyte chemoattractant protein-1 gene expression in prostatic hyperplasia and prostate adenocarcinoma.  

PubMed Central

Human monocyte chemoattractant protein-1 (MCP-1) has been shown to act as a chemokine in the recruitment of monocyte/macrophages during inflammation states. Furthermore, there is increasing evidence that MCP-1 is involved in the recruitment of tumor-associated macrophages. In vivo, one of the major cellular sources of MCP-1 are the smooth muscle cells. As MCP-1 gene expression and/or protein production in these cells is not necessarily correlated with the accumulation of inflammatory cells, there might possibly be additional functions of this cytokine. In the present study, we investigated by use of 35S-labeled antisense RNA probes whether the MCP-1 gene is expressed in tissue specimens of benign prostatic hyperplasia (n = 13) and specimens of prostate carcinoma (n = 8), both of which are characterized by a prominent fibromuscular stroma and inconspicuous inflammatory infiltrates. MCP-1 transcripts were located in stromal smooth muscle cells and, additionally, in basal cells of benign prostatic glands. In prostate carcinoma, the number of MCP-1 mRNA-expressing cells was significantly less than in benign prostatic hyperplasia. MCP-1 transcripts were located in preserved fibromuscular stroma and in basal cells of entrapped non-neoplastic glands but not in carcinomatous cells. Immunohistochemical staining with polyclonal antibodies raised against MCP-1 revealed strong reactivity in the fibromuscular stroma surrounding both benign and malignant glands. MCP-1 gene expression or immunoreactivity for anti-MCP-1 antibodies was not related to the rare, lymphocytic interstitial infiltrates. The results show that 1) in the absence of significant leukocyte accumulation, it is unlikely that MCP-1 exerts chemotactic functions in the prostate and 2) that MCP-1, in contrast to previous findings in a wide variety of other human neoplasms, is not expressed in carcinomatous cells of the prostate. Images Figure 2 Figure 3 Figure 4

Mazzucchelli, L.; Loetscher, P.; Kappeler, A.; Uguccioni, M.; Baggiolini, M.; Laissue, J. A.; Mueller, C.



HIV-1 Induces Telomerase Activity in Monocyte-Derived Macrophages, Possibly Safeguarding One of Its Reservoirs  

PubMed Central

Monocyte-derived macrophages (MDM) are widely distributed in all tissues and organs, including the central nervous system, where they represent the main part of HIV-infected cells. In contrast to activated CD4+ T lymphocytes, MDM are resistant to cytopathic effects and survive HIV infection for a long period of time. The molecular mechanisms of how HIV is able to persist in macrophages are not fully elucidated yet. In this context, we have studied the effect of in vitro HIV-1 infection on telomerase activity (TA), telomere length, and DNA damage. Infection resulted in a significant induction of TA. This increase was directly proportional to the efficacy of HIV infection and was found in both nuclear and cytoplasmic extracts, while neither UV light-inactivated HIV nor exogenous addition of the viral protein Tat or gp120 affected TA. Furthermore, TA was not modified during monocyte-macrophage differentiation, MDM activation, or infection with vaccinia virus. HIV infection did not affect telomere length. However, HIV-infected MDM showed less DNA damage after oxidative stress than noninfected MDM, and this resistance was also increased by overexpressing telomerase alone. Taken together, our results suggest that HIV induces TA in MDM and that this induction might contribute to cellular protection against oxidative stress, which could be considered a viral strategy to make macrophages better suited as longer-lived, more resistant viral reservoirs. In the light of the clinical development of telomerase inhibitors as anticancer therapeutics, inhibition of TA in HIV-infected macrophages might also represent a novel therapeutic target against viral reservoirs.

Reynoso, Rita; Wieser, Matthias; Ojeda, Diego; Bonisch, Maximilian; Kuhnel, Harald; Bolcic, Federico; Quendler, Heribert; Grillari, Johannes



C-Reactive Protein in the Arterial Intima Role of C-Reactive Protein Receptor-Dependent Monocyte Recruitment in Atherogenesis  

Microsoft Academic Search

Infiltration of monocytes into the arterial wall is an early cellular event in atherogenesis. Recent evidence shows that C-reactive protein (CRP) is deposited in the arterial intima at sites of atherogenesis. In this study, we demonstrate that CRP deposition precedes the appearance of monocytes in early atherosclerotic lesions. CRP is chemotactic for freshly isolated human blood monocytes. A specific CRP

Michael Torzewski; Carsten Rist; Richard F. Mortensen; Thomas P. Zwaka; Magda Bienek; Johannes Waltenberger; Wolfgang Koenig; Gerd Schmitz; Vinzenz Hombach; Jan Torzewski



Regulation of human monocyte/macrophage function by extracellular matrix. Adherence of monocytes to collagen matrices enhances phagocytosis of opsonized bacteria by activation of complement receptors and enhancement of Fc receptor function.  

PubMed Central

In inflammation monocytes emigrate from the peripheral circulation into an extravascular area rich in extracellular matrix proteins. In this milieu, phagocytes ingest and kill invading pathogens. In the present studies, we found that monocytes adhered to type I collagen gels phagocytized 2.5-12-fold more opsonized Escherichia coli, Staphylococcus aureus, Streptococcus pyogenes, and Streptococcus pneumoniae than plastic-adherent monocytes. The rate of phagocytosis and the number of bacteria ingested by collagen-adherent monocytes was equal to, or greater than, the number of bacteria ingested by 7-d cultured macrophages (M phi). Although both collagen- and plastic-adherent monocytes were bactericidal for E. coli and S. aureus, more bacteria were killed by collagen-adherent monocytes by virtue of their enhanced phagocytic capacity. Cultured M phi only were bacteriostatic. Adherence of monocytes to collagen gels activated C receptors (CR) types 1 and 3 for phagocytosis, and enhanced Fc receptor (FcR)-mediated phagocytosis. Collagen- and plastic-adherent monocytes produced equivalent amounts of superoxide anion in response to phorbol myristate acetate and opsonized zymosan. Thus, the enhanced phagocytosis and killing of opsonized bacteria by collagen-adherent monocytes appear to be by regulation of the function of membrane CR and FcR, without apparent enhancement of the respiratory burst. These data suggest that adherence of monocytes to the extracellular matrix during inflammation may rapidly activate these cells for enhanced phagocytic bactericidal activity.

Newman, S L; Tucci, M A



Cytokine Pattern of T Lymphocytes in Acute Schistosomiasis mansoni Patients following Treated Praziquantel Therapy.  


Acute schistosomiasis is associated with a primary exposure and is more commonly seen in nonimmune individuals traveling through endemic regions. In this study, we have focused on the cytokine profile of T lymphocytes evaluated in circulating leukocytes of acute Schistosomiasis mansoni-infected patients (ACT group) before and after praziquantel treatment (ACT-TR group). Our data demonstrated increased values of total leukocytes, eosinophils, and monocytes in both groups. Interestingly, we have observed that patients treated with praziquantel showed increased values of lymphocytes as compared with noninfected group (NI) or ACT groups. Furthermore, a decrease of neutrophils in ACT-TR was observed when compared to ACT group. Analyses of short-term in vitro whole blood stimulation demonstrated that, regardless of the presence of soluble Schistosoma mansoni eggs antigen (SEA), increased synthesis of IFN-? and IL-4 by T-cells was observed in the ACT group. Analyses of cytokine profile in CD8 T cells demonstrated higher percentage of IFN-? and IL-4 cells in both ACT and ACT-TR groups apart from increased percentage of IL-10 cells only in the ACT group. This study is the first one to point out the relevance of CD8 T lymphocytes in the immune response induced during the acute phase of schistosomiasis. PMID:23401741

Silveira-Lemos, Denise; Fernandes Costa-Silva, Matheus; Cardoso de Oliveira Silveira, Amanda; Azevedo Batista, Mauricio; Alves Oliveira-Fraga, Lúcia; Soares Silveira, Alda Maria; Barbosa Alvarez, Maria Carolina; Martins-Filho, Olindo Assis; Gazzinelli, Giovanni; Corrêa-Oliveira, Rodrigo; Teixeira-Carvalho, Andréa



Cytokine Pattern of T Lymphocytes in Acute Schistosomiasis mansoni Patients following Treated Praziquantel Therapy  

PubMed Central

Acute schistosomiasis is associated with a primary exposure and is more commonly seen in nonimmune individuals traveling through endemic regions. In this study, we have focused on the cytokine profile of T lymphocytes evaluated in circulating leukocytes of acute Schistosomiasis mansoni-infected patients (ACT group) before and after praziquantel treatment (ACT-TR group). Our data demonstrated increased values of total leukocytes, eosinophils, and monocytes in both groups. Interestingly, we have observed that patients treated with praziquantel showed increased values of lymphocytes as compared with noninfected group (NI) or ACT groups. Furthermore, a decrease of neutrophils in ACT-TR was observed when compared to ACT group. Analyses of short-term in vitro whole blood stimulation demonstrated that, regardless of the presence of soluble Schistosoma mansoni eggs antigen (SEA), increased synthesis of IFN-? and IL-4 by T-cells was observed in the ACT group. Analyses of cytokine profile in CD8 T cells demonstrated higher percentage of IFN-? and IL-4 cells in both ACT and ACT-TR groups apart from increased percentage of IL-10 cells only in the ACT group. This study is the first one to point out the relevance of CD8 T lymphocytes in the immune response induced during the acute phase of schistosomiasis.

Silveira-Lemos, Denise; Fernandes Costa-Silva, Matheus; Cardoso de Oliveira Silveira, Amanda; Azevedo Batista, Mauricio; Alves Oliveira-Fraga, Lucia; Soares Silveira, Alda Maria; Barbosa Alvarez, Maria Carolina; Martins-Filho, Olindo Assis; Gazzinelli, Giovanni; Correa-Oliveira, Rodrigo; Teixeira-Carvalho, Andrea



Distinct functions of chemokine receptor axes in the atherogenic mobilization and recruitment of classical monocytes.  


We used a novel approach of cytostatically induced leucocyte depletion and subsequent reconstitution with leucocytes deprived of classical (inflammatory/Gr1(hi) ) or non-classical (resident/Gr1(lo) ) monocytes to dissect their differential role in atheroprogression under high-fat diet (HFD). Apolipoprotein E-deficient (Apoe(-/-) ) mice lacking classical but not non-classical monocytes displayed reduced lesion size and macrophage and apoptotic cell content. Conversely, HFD induced a selective expansion of classical monocytes in blood and bone marrow. Increased CXCL1 levels accompanied by higher expression of its receptor CXCR2 on classical monocytes and inhibition of monocytosis by CXCL1-neutralization indicated a preferential role for the CXCL1/CXCR2 axis in mobilizing classical monocytes during hypercholesterolemia. Studies correlating circulating and lesional classical monocytes in gene-deficient Apoe(-/-) mice, adoptive transfer of gene-deficient cells and pharmacological modulation during intravital microscopy of the carotid artery revealed a crucial function of CCR1 and CCR5 but not CCR2 or CX3 CR1 in classical monocyte recruitment to atherosclerotic vessels. Collectively, these data establish the impact of classical monocytes on atheroprogression, identify a sequential role of CXCL1 in their mobilization and CCR1/CCR5 in their recruitment. PMID:23417922

Soehnlein, Oliver; Drechsler, Maik; Döring, Yvonne; Lievens, Dirk; Hartwig, Helene; Kemmerich, Klaus; Ortega-Gómez, Almudena; Mandl, Manuela; Vijayan, Santosh; Projahn, Delia; Garlichs, Christoph D; Koenen, Rory R; Hristov, Mihail; Lutgens, Esther; Zernecke, Alma; Weber, Christian



Monocytes from systemic lupus erythematous patients are severely altered in phenotype and lineage flexibility  

PubMed Central

OBJECTIVE—Cells of the myeloid lineage comprise a very heterogeneous population with many phenotypes and functional activities including macrophages and dendritic cells. To investigate the status, differentiative potential and lineage commitment of monocytic cells in systemic lupus erythematosus (SLE) patients, this study isolated and cultured peripheral blood monocytes from patients and healthy donors. ?METHODS—Monocytes were isolated by gradient centrifugation and adherence to plastic dishes. The cells were then cultured for three days, partially supplemented with GM-CSF and interleukin 4 (IL4) to obtain dendritic cells. The differentiation status was monitored by the expression of surface markers using flow cytometry and cytokine secretion.?RESULTS—Monocytes from SLE patients expressed significantly lower numbers of the monocytic marker CD14 and HLA-DR while secreting significantly more tumour necrosis factor ? (TNF?) than monocytes from healthy donors. The addition of GM-CSF and IL4 resulted in an inhibition of TNF? secretion, but was not sufficient to generate monocytederived dendritic cells.?CONCLUSION—Monocytes from SLE patients are severely altered in phenotype and function and have a limited differentiation flexibility towards the accessory type of monocytic cells.??

Steinbach, F.; Henke, F.; Krause, B.; Thiele, B.; Burmester, G.; Hiepe, F.



Fatty acids from VLDL lipolysis products induce lipid droplet accumulation in human monocytes  

PubMed Central

One mechanism by which monocytes become activated postprandially is by exposure to triglyceride (TG)-rich lipoproteins such as very low-density lipoproteins (VLDL). VLDL are hydrolyzed by lipoprotein lipase (LpL) at the blood-endothelial cell interface, releasing free fatty acids. In this study, we examined postprandial monocyte activation in more detail, and found that lipolysis products generated from postprandial VLDL induce the formation of lipid-filled droplets within cultured THP-1 monocytes, characterized by coherent anti-stokes Raman spectroscopy. Organelle-specific stains revealed an association of lipid droplets with the endoplasmic reticulum, confirmed by electron microscopy. Lipid droplet formation was reduced when LpL-released fatty acids were bound by bovine serum albumin, which also reduced cellular inflammation. Furthermore, saturated fatty acids induced more lipid droplet formation in monocytes compared to mono- and polyunsaturated fatty acids. Monocytes treated with postprandial VLDL lipolysis products contained lipid droplets with more intense saturated Raman spectroscopic signals than monocytes treated with fasting VLDL lipolysis products. In addition, we found that human monocytes isolated during the peak postprandial period contain more lipid droplets compared to those from the fasting state, signifying that their development is not limited to cultured cells but also occurs in vivo. In summary, circulating free fatty acids can mediate lipid droplet formation in monocytes and potentially be used as a biomarker to assess an individual’s risk of developing atherosclerotic cardiovascular disease.

den Hartigh, Laura J; Connolly-Rohrbach, Jaime E; Fore, Samantha; Huser, Thomas R; Rutledge, John C



Differential Responses Between Monocytes and Monocyte-Derived Macrophages for Lipopolysaccharide Stimulation of Calves  

Microsoft Academic Search

In this experiment Toll-like receptor expression pattern in monocytes and monocyte-derived macrophages by lipopolysaccharide (LPS) stimulation was examined. Jugular venous blood was collected from four Japanese calves, and the peripheral blood mononuclear cells (PBMCs) were isolated. The cells were directly used for collecting monocytes by magnetic cell sorting or cultured for 7 days to collect monocyte-derived macrophages in Repcell. Then

Yijie Guo; Guoqi Zhao; Sachi Tanaka; Takahiro Yamaguchi



The CD19/CD81 complex physically interacts with CD38 but is not required to induce proliferation in mouse B lymphocytes  

PubMed Central

In B lymphocytes, the cell surface receptor CD38 is involved in apoptosis of immature B cells, proliferation and differentiation of mature B cells. Although CD38 has been establish as a receptor, its signaling has been only partially characterized. As a result of the lack of signaling motifs in the cytoplasmic domain, CD38 must use a co-receptor to induce signaling within the cell. Accordingly, CD38 has been associated with different receptors such as the T-cell receptor/CD3 complex on T cells, CD16 on natural killer cells and MHC class II molecules on monocytes. The CD19/CD81 complex has been proposed as a co-receptor for CD38 in human B lymphocytes, but little or no characterization has been performed in mice. In this study the contribution of the CD19/CD81 complex in murine CD38 signaling was evaluated. Proliferation assays were performed using CD19?/? or CD81?/? deficient mice; CFSE-labeled B lymphocytes from wild-type mice and CD19?/?, CD81?/? and CD38?/? deficient mice were stimulated with agonistic antibodies against CD38. Immunoprecipitation and immunofluorescence were also performed to detect protein–protein interactions. Our results indicate that the CD19/CD81 complex interacts with CD38 but this interaction is not required to induce proliferation in mouse B lymphocytes, suggesting that other receptors may contribute to the proliferation induced by CD38 in B lymphocytes.

Vences-Catalan, Felipe; Rajapaksa, Ranjani; Levy, Shoshana; Santos-Argumedo, Leopoldo



Lymphocyte count was significantly associated with hyper-LDL cholesterolemia independently of high-sensitivity C-reactive protein in apparently healthy Japanese  

Microsoft Academic Search

The aim of this study was to investigate the association between leukocyte subtype counts and hyper-LDL cholesterolemia, hypertriglyceridemia,\\u000a and hypo-HDL cholesterolemia. Logistic regressions using hyper-LDL cholesterolemia, hypertriglyceridemia, and hypo-HDL cholesterolemia\\u000a as a dependent variable and total leukocyte, basophil, eosinophil, neutrophil, lymphocyte, and monocyte counts as an independent\\u000a variable were calculated adjusting for age, body mass index (BMI), high-sensitivity C-reactive protein

Eiji Oda; Ryu Kawai; Yoshifusa Aizawa


Lymphocyte Adhesion and Interactions with Biomaterial Adherent Macrophages and Foreign Body Giant Cells  

PubMed Central

To characterize the effects of adherent macrophages and biomaterial surface chemistries on lymphocyte adhesion and activation, lymphocytes were co-cultured with monocytes alone and together, directly and separated by a porous membrane transwell on hydrophobic, hydrophilic/neutral, hydrophilic/anionic, and hydrophilic/cationic biomaterial surfaces. Surface adherent cells were quantitatively analyzed after 3 days utilizing immunofluorescence and phase contrast imaging. After periods of 3, 7, and 10 days, secreted interferon-? (IFN-?) was quantified by ELISA. Limited direct biomaterial-adherent lymphocytes were identified regardless of the presence of macrophages or foreign body giant cells (FBGC). The majority of adherent lymphocytes, which were T cells (> 95%) rather than natural killer cells, predominantly interacted with adherent macrophages and FBGCs; greater than 90% were interacting on surfaces with higher levels of adherent macrophages and FBGCs and greater than 55% were interacting on surfaces with lower levels of macrophages and FBGCs. The hydrophilic/anionic surface promoted higher levels of macrophage- and FBGC-adherent lymphocytes but was nonselective for lymphocyte subtype interactions. The hydrophilic/neutral surface was selective for CD4+ T lymphocyte interactions while the hydrophobic surface was selective for CD8+ T lymphocyte interactions. IFN-? was produced in direct and indirect co-cultures but not in lymphocyte- and monocyte-only cultures suggesting that lymphocytes are activated via macrophage-derived cytokines rather than direct biomaterial contact. Direct lymphocyte interactions with adherent macrophages/FBGCs enhanced IFN-? production relative to indirect co-cultures. These results suggest that lymphocytes prefer interactions with adherent macrophages and FBGCs, resulting in lymphocyte activation, and these interactions can be influenced by biomaterial surface chemistries.

Chang, David T.; Colton, Erica; Matsuda, Takehisa; Anderson, James M.



p53 Expression in circulating lymphocytes of non-melanoma skin cancer patients from an arsenic contaminated region in Mexico. A pilot study  

Microsoft Academic Search

Arsenic is a common environmental toxicant and epidemiological studies associate arsenic exposure with various pathologic disorders and several types of cancer. Skin cancers are the most common arsenic-induced neoplasias and the prevalence of skin lesions has been reported to be significantly elevated in individuals exposed to arsenic via drinking water in México. Being lymphocytes the main cells used for human

Ana M. Salazar; Emma Calderón-Aranda; Mariano E. Cebrián; Monserrat Sordo; Andrés Bendesky; Arístides Gómez-Muñoz; Leonor Acosta-Saavedra; Patricia Ostrosky-Wegman



?? T Lymphocytes Coordinate Eosinophil Influx during Allergic Responses  

PubMed Central

Tissue eosinophil infiltration, which is a hallmark of allergic and helminthic diseases, is mainly coordinated by T lymphocytes, via the production of eosinophilotactic chemokines. Among T lymphocyte subsets, lymphocytes expressing ?? T cell receptor have been determined as a key factor for eosinophil accumulation via direct and indirect mechanisms. This knowledge is strongly supported by the fact that, in different experimental models of eosinophilic airway inflammation and helminth-induced Th2 lung inflammation, an evident tissue accumulation of ?? T lymphocytes is observed. In addition, the depletion of ?? T lymphocytes is correlated with the impairment of eosinophil accumulation in inflamed tissue. ?? T lymphocytes are non-conventional T lymphocytes, which comprise a minor T lymphocyte subset, mainly distributed in the tissue, and present crucial roles in innate and acquired immune responses. ?? T lymphocytes recognize several danger- and pathogen-associated molecular pattern molecules and stress antigens in a MHC-independent fashion and can provide rapid tissue-specific responses, via the production of a wide range of chemical mediators capable to modulate other cell populations. These mediators include chemoattractant cytokines and chemokines that attract eosinophils into the tissue by either direct recognition (such as IL-5, CCL11/eotaxin), or indirect mechanisms via the modulation of ?? T lymphocytes and macrophages (through the production of interferon-?, IL-4, and CCL2/Monocyte chemoattractant protein-1, MCP-1, for example). The present review presents an overview of how ?? T lymphocytes coordinate eosinophil accumulation in allergy, by focusing on their role in airway inflammation and by discussing the involvement of cytokines and chemokines in this phenomenon.

de Oliveira Henriques, Maria Das Gracas Muller; Penido, Carmen



Carboxylesterase 1 (Ces1): from monocyte marker to major player  

Microsoft Academic Search

There are few, if any, enzymes that have been studied by, and have importance in, so many and varied disciplines as has monocyte esterase\\/Ces 1. In this review its involvement in the fields of histochemistry, haematology, toxicology, pharmacology, therapeutics, and tumour cell killing, immune surveillance and malignant disease, is briefly described.

Geraldine M Markey




Technology Transfer Automated Retrieval System (TEKTRAN)

Recent studies have shown that mice fed soy-based diets exhibited reduced atherosclerotic lesions. However, the mechanism(s) of soy-mediated atheroprotection is not known. Since, interaction of endothelial cells and monocytes is an initial event in the development of atherosclerosis, we hypothesiz...


Monocyte-derived dendritic cells in innate and adaptive immunity  

Microsoft Academic Search

Monocytes have been classically considered essential elements in relation with innate immune responses against pathogens, and inflammatory processes caused by external aggressions, infection and autoimmune disease. However, although their potential to differentiate into dendritic cells (DCs) was discovered 14 years ago, their functional relevance with regard to adaptive immune responses has only been uncovered very recently. Studies performed over the

Beatriz León; Carlos Ardavín



Chemotactic activities of avian lymphocytes.  


Chicken lymphocytes, enriched chicken T and B lymphocytes, and a turkey B-lymphoblastoid cell line, the RP-9 cells, are used in the studies of chemotaxis in a Boyden-type chamber assay. The chemoattractants used are lipopolysaccharide, fMLP, interleukin-8, MIP1-beta, rabbit anti-chicken IgG and IgM. The results indicate that all these cells can migrate into the polycarbonate membranes in the absence of chemoattractants. When the chemoattractants are present, the numbers of migrating cells are greatly increased. It is, therefore, concluded that avian lymphocytes have the ability to migrate, and can respond to chemical signals which result in chemotaxis and accumulation of lymphocytes at the sites where the signals originate. PMID:10579392

Lam, K M


Monocyte-to-Macrophage Differentiation  

PubMed Central

Although monocyte- and macrophage-derived molecules are known to promote extracellular matrix (ECM) disruption and destabilization, it is less appreciated that they also synthesize molecules contributing to ECM formation, stabilization, and function. We have identified and characterized the synthesis of proteoglycans and related proteins, some not previously known to be associated with macrophages. Proteoglycan extracts of [35S]sulfate- and 35S-trans amino acid-radiolabeled culture media from THP-1 monocytes induced to differentiate by treatment with phorbol myristate acetate revealed three major proteins of ?25, 90, and 100 kDa following chondroitin ABC lyase digestion. The 25-kDa protein was predominant for monocytes, whereas the 90- and 100-kDa proteins were predominant for macrophages. Tandem mass spectrometry identified (i) the 25-kDa core protein as serglycin, (ii) the 90-kDa core protein as inter-?-inhibitor heavy chain 2 (I?IHC2), and (iii) the 100-kDa core as amyloid precursor-like protein 2 (APLP2). Differentiation was also associated with (i) a >500-fold increase in mRNA for TNF-stimulated gene-6, an essential cofactor for heavy chain-mediated matrix stabilization; (ii) a >800-fold increase in mRNA for HAS2, which is responsible for hyaluronan synthesis; and (iii) a 3-fold increase in mRNA for versican, which interacts with hyaluronan. Biochemical evidence is also presented for an I?IHC2-APLP2 complex, and immunohistochemical staining of human atherosclerotic lesions demonstrates similar staining patterns for APLP2 and I?IHC2 with macrophages, whereas serglycin localizes to the underlying glycosaminoglycan-rich region. These findings indicate that macrophages synthesize many of the molecules participating in ECM formation and function, suggesting a novel role for these molecules in the differentiation of macrophages in the development of atherosclerosis.

Chang, Mary Y.; Chan, Christina K.; Braun, Kathleen R.; Green, Pattie S.; O'Brien, Kevin D.; Chait, Alan; Day, Anthony J.; Wight, Thomas N.



HSV-2 regulates monocyte inflammatory response via the Fas/FasL pathway.  


Monocytic cells represent important cellular elements of the innate and adaptive immune responses in viral infections. We assessed the role of Fas/FasL in promoting monocyte apoptosis during HSV-2 infection by using an in vitro model based on the murine RAW 264.7 monocytic cell line and an in vivo murine model of HSV-2 infection applied to C57BL6, MRL-Fas(lpr)/J (Fas-/-) and C3-Fasl(gld)/J (FasL-/-) mice. HSV-2 infection of the monocytic cell line led to early induction of apoptosis, with no protective expression of anti-apoptotic Bcl-2. HSV-2 infected monocytes up-regulated Fas and FasL expression early during in vitro infection but were susceptible to Fas induced apoptosis. The vaginal monocytes in the HSV-2 murine model of infection up-regulated FasL expression and were susceptible to Fas induced apoptosis. HSV-2 infection of Fas and FasL- deficient mice led to decreased apoptosis of monocytes and impaired recruitment of NK, CD4+ and CD8+ T cells within the infection sites. The vaginal lavages of HSV-2 infected Fas and FasL- deficient showed decreased production of CXCL9, CXCL10 and TNF-? in comparison to HSV-2 infected wild-type mice strain. The decreased recruitment of immune competent cells was accompanied by delayed virus clearance from the infected tissue. Triggering of the Fas receptor on HSV-2 infected monocytes in vitro up-regulated the expression of CXCL9 chemokines and the cytokine TNF-?. Our study provides novel insights on the role of Fas/FasL pathway not only in apoptosis of monocytes but also in regulating local immune response by monocytes during HSV-2 infection. PMID:23922974

Krzyzowska, Malgorzata; Baska, Piotr; Orlowski, Piotr; Zdanowski, Robert; Winnicka, Anna; Eriksson, Kristina; Stankiewicz, Wanda



HSV-2 Regulates Monocyte Inflammatory Response via the Fas/FasL Pathway  

PubMed Central

Monocytic cells represent important cellular elements of the innate and adaptive immune responses in viral infections. We assessed the role of Fas/FasL in promoting monocyte apoptosis during HSV-2 infection by using an in vitro model based on the murine RAW 264.7 monocytic cell line and an in vivo murine model of HSV-2 infection applied to C57BL6, MRL-Faslpr/J (Fas?/?) and C3-Faslgld/J (FasL?/?) mice. HSV-2 infection of the monocytic cell line led to early induction of apoptosis, with no protective expression of anti-apoptotic Bcl-2. HSV-2 infected monocytes up-regulated Fas and FasL expression early during in vitro infection but were susceptible to Fas induced apoptosis. The vaginal monocytes in the HSV-2 murine model of infection up-regulated FasL expression and were susceptible to Fas induced apoptosis. HSV-2 infection of Fas and FasL- deficient mice led to decreased apoptosis of monocytes and impaired recruitment of NK, CD4+ and CD8+ T cells within the infection sites. The vaginal lavages of HSV-2 infected Fas and FasL- deficient showed decreased production of CXCL9, CXCL10 and TNF-? in comparison to HSV-2 infected wild-type mice strain. The decreased recruitment of immune competent cells was accompanied by delayed virus clearance from the infected tissue. Triggering of the Fas receptor on HSV-2 infected monocytes in vitro up-regulated the expression of CXCL9 chemokines and the cytokine TNF-?. Our study provides novel insights on the role of Fas/FasL pathway not only in apoptosis of monocytes but also in regulating local immune response by monocytes during HSV-2 infection.

Krzyzowska, Malgorzata; Baska, Piotr; Orlowski, Piotr; Zdanowski, Robert; Winnicka, Anna; Eriksson, Kristina; Stankiewicz, Wanda



Comparative Cytokine Release from Human Monocytes, Monocyte-Derived Immature Mast Cells, and a Human Mast Cell Line (HMC1)  

Microsoft Academic Search

To obtain further information regarding the role of cytokines during mast cell differentiation, we have investigated changes of cytokine secretion in mast cells developing from the human peripheral blood monocytic cell fraction during culture with fibroblast- derived conditioned media. The influence of stem cell factor and an antibody to the respective receptor in our culture system was studied as well.Interleukin

Jürgen Grabbe; Pia Welker; Annelie Möller; Edgar Dippel; Leonie K. Ashman; Beate M. Czarnetzki



Adrenaline Inhibits Lipopolysaccharide-Induced Macrophage Inflammatory Protein1?? in Human Monocytes: The Role of ??Adrenergic Receptors  

Microsoft Academic Search

Macrophage inflammatory protein-1 (MIP-1) has an important role in the development of inflammatory re- sponses during infection by regulating leukocyte traf- ficking and function. Our study was conducted to in- vestigate the effect of adrenaline on lipopolysaccharide (LPS)-induced MIP-1 production by human periph- eral blood monocytes and human monocytic THP-1 cells. Monocytes were incubated in vitro with LPS for 4

Chi-Yuan Li; Tz-Chong Chou; Chian-Her Lee; Chien-Sung Tsai; Shih-Hurng Loh; Chih-Shung Wong



Bone Morphogenic Protein7 Inhibits Monocyte-Stimulated TGF-1 Generation in Renal Proximal Tubular Epithelial Cells  

Microsoft Academic Search

It has been demonstrated that bone morphogenic protein-7 (BMP-7) stimulates formation of hyaluronan (HA)-based cables on the cell surface of renal proximal tubular cells and that these cables mediate monocyte binding. Furthermore, interaction of monocytes with proximal tubule cell (PTC) surface intracellular adhesion molecule (ICAM) stimulates the synthesis of TGF-1. This study examined the effect of BMP-7 on monocyte-stimulated TGF-1

Xiao Liang Zhang; Wisam Selbi; Carol de la Motte; Vincent Hascall; Aled O. Phillips; Cardiff Wales



Induction of Monocyte Chemoattractant Protein1 in HIV1 Tat-Stimulated Astrocytes and Elevation in AIDS Dementia  

Microsoft Academic Search

Activated monocytes release a number of substances, including inflammatory cytokines and eicosanoids, that are highly toxic to cells of the central nervous system. Because monocytic infiltration of the central nervous system closely correlates with HIV-1-associated dementia, it has been suggested that monocyte-derived toxins mediate nervous system damage. In the present study, we show that the HIV-1 transactivator protein Tat significantly

Katherine Conant; Alfredo Garzino-Demo; Avindra Nath; Justin C. McArthur; William Halliday; Christopher Power; Robert C. Gallo



P48-Triggered transmembrane signaling transduction of human monocytes: mobilization of calcium ion and activation of protein kinase C (PKC)  

Microsoft Academic Search

P48 is a cytokine which induces monocyte differentiation and the induction of cytotoxic activity. In this study, the signal transduction events involved in the stimulation of monocytes with the membrane form of P48 (mP48) were investigated. Monocyte stimulation with mP48 was found to involve the mobilization of intracellular calcium (Ca2+) and the activation and translocation of PKC from the cytosol

ZL Chang; Re Hall



3D Porous Chitosan-Alginate Scaffolds: New Matrix for Studying Prostate Cancer Cell-Lymphocyte Interactions In Vitro  

PubMed Central

The treatment of castration-resistant prostate cancer (CRPC) remains palliative. Immunotherapy offers a potentially effective therapy for CRPC; however, its advancement into the clinic has been slow, in part because of the lack of representative in vitro tumor models that resemble the in vivo tumor microenvironment for studying interactions of CRPC cells with immune cells and other potential therapeutics. This study evaluates the use of 3D porous chitosan-alginate (CA) scaffolds for culturing human prostate cancer (PCa) cells and studying tumor cell interaction with human peripheral blood lymphocytes (PBLs) ex vivo. CA scaffolds and Matrigel matrix samples supported in vitro tumor spheroid formation over 15 days of culture, and CA scaffolds supported live cell fluorescence imaging with confocal microscopy using stably transfected PCa cells for 55 days. PCa cells grown in Matrigel matrix and CA scaffolds for 15 days were co-cultured with PBLs for 2 and 6 days in vitro and evaluated with scanning electron microscopy (SEM), immunohistochemistry (IHC), and flow cytometry. Both the Matrigel matrix and CA scaffolds supported interaction of PBLs with PCa tumors, with CA scaffolds providing a more robust platform for subsequent analyses. This study demonstrates the use of 3D natural polymer scaffolds as a tissue culture model for supporting long-term analysis of interaction of prostate cancer tumor cells with immune cells, providing an in vitro platform for rapid immunotherapy development.

Florczyk, Stephen J.; Liu, Gang; Kievit, Forrest M.; Lewis, Allison M.; Wu, Jennifer D.



Effect of He-Ne laser irradiation on erythrocyte and lymphocyte membranes of children in vitro as studied by the intrinsic and extrinsic fluorescence techniques  

NASA Astrophysics Data System (ADS)

In recent years the treatment of blood with low intensity laser irradiation has become popular in a variety of clinical applications due to its anti-inflammatory, biostimulative and immune-stimulatory effects etc. Despite of wide using of laser blood irradiation in the pediatric practice there is lack of information concerning the sensitivity of children blood cells to laser irradiation. At present study the influence of the He-Ne laser irradiation on the lipid physico-chemical state in lymphocytes and isolated erythrocyte membranes of 8-16 years old children using lipophilic fluorescence probe pyrene was investigated in vitro. It was shown that fluorescence parameters of pyrene incorporated into erythrocyte and lymphocyte membranes after laser irradiation ((lambda) equals630nm) at dose of 24 J/cm2 at tequals18+/- 2 degree(s)C were unchanged. The intensity of intrinsic protein UV-fluorescence ((lambda) ex equals 297 nm, $lamdaem equals 332 nm) of lymphocytes exposed to the same irradiation was decreased insignificantly. The obtained data indicate that He-Ne laser irradiation at the above dose does not affect the lipid microviscosity of erythrocyte and lymphocyte membranes.

Volotovskaya, Anna V.; Kozlova, Nataly M.; Slobozhanina, Ekaterina I.; Ulaschik, Vladimir S.; Mostovnikov, Vasili A.



Interaction between human peripheral blood monocytes and tumor promoters: Effect on growth differentiation and function in vitro  

SciTech Connect

Studies on the differentiation and activation of human monocytes in tissue cultures have usually been limited by the deterioration of human monocytes and macrophages in long-term cultures. In this study, we attempted to establish long-term human monocyte/macrophage cultures using the phorbol ester 12-0 tetradecanoyl-phorbol-13-acetate (TPA), and we studied the morphology, function, and biochemical properties of such treated human blood monocytes. Enriched suspensions of monocytes were obtained using Ficoll-Hypaque gradient and cultured in the absence or presence of various concentrations of TPA. Samples were removed at different times and processed for scanning electron microscopy. Parallel samples were examined for numbers of adherent cells, phagocytosis, oxidative burst, beta-galactosidase assays, and lectin-mediated erythrolysis. TPA-treated monocytes survived in larger numbers in culture for up to 7 weeks and were more pleomorphic and exhibited higher beta-galactosidase activities after 14 days in culture than untreated monocytes. TPA-treated cells and untreated cells in long-term cultures showed a decrease in their oxidative burst activity while their phagocytic activity was not affected, and the TPA treatment augmented the lysis of wheat germ agglutinin-opsonized erythrocytes by the cultured monocytes. TPA treatment of adherent human monocytes resulted in cell cultures with increased numbers of viable and functionally adherent cells for extended periods of time and does not seem to interfere with the differentiation and maturation of the cells in culture.

Keisari, Y.; Bucana, C.; Markovich, S.; Campbell, D.E. (Tel Aviv Univ. (Israel))



Characterization of a New Chronic Lymphocytic Leukemia Cell Line for Mechanistic In Vitro and In Vivo Studies Relevant to Disease  

PubMed Central

Studies of chronic lymphocytic leukemia (CLL) have yielded substantial progress, however a lack of immortalized cell lines representative of the primary disease has hampered a full understanding of disease pathogenesis and development of new treatments. Here we describe a novel CLL cell line (OSU-CLL) generated by EBV transformation, which displays a similar cytogenetic and immunophenotype observed in the patient’s CLL (CD5 positive with trisomy 12 and 19). A companion cell line was also generated from the same patient (OSU-NB). This cell line lacked typical CLL characteristics, and is likely derived from the patient’s normal B cells. In vitro migration assays demonstrated that OSU-CLL exhibits migratory properties similar to primary CLL cells whereas OSU-NB has significantly reduced ability to migrate spontaneously or towards chemokine. Microarray analysis demonstrated distinct gene expression patterns in the two cell lines, including genes on chromosomes 12 and 19, which is consistent with the cytogenetic profile in this cell line. Finally, OSU-CLL was readily transplantable into NOG mice, producing uniform engraftment by three weeks with leukemic cells detectable in the peripheral blood spleen and bone marrow. These studies describe a new CLL cell line that extends currently available models to study gene function in this disease.

Hertlein, Erin; Beckwith, Kyle A.; Lozanski, Gerard; Chen, Timothy L.; Towns, William H.; Johnson, Amy J.; Lehman, Amy; Ruppert, Amy S.; Bolon, Brad; Andritsos, Leslie; Lozanski, Arletta; Rassenti, Laura; Zhao, Weiqiang; Jarvinen, Tiina M.; Senter, Leigha; Croce, Carlo M.; Symer, David E.; de la Chapelle, Albert; Heerema, Nyla A.; Byrd, John C.



Bovine lymphocytic leukemia: studies of etiology, pathogenesis and mode of transmission. Progress report, November 1, 1979-October 31, 1980  

SciTech Connect

The primary project objectives are to elucidate the cause(s) and early pathogenesis of the adult form of lymphosarcoma in cattle. This goal is to be accomplished through experimental transmission of the disease. For these studies large quantities of bovine leukemia virus (BLV) were propagated in short-term, mitogen stimulated, lymphocyte cultures. Cultures containing abundant BLV particles were pooled (33 liters total) and further processed by continuous flow, density gradient, ultracentrifugation. This higly concentrated, cell free, BLV preparation was then used as inoculum for 12 late stage bovine fetuses (inoculated in utero) and two newborn calves. Extensive monitoring studies have been carrid out on these inoculated animals to detect precancerous changes and to obtain a detailed description of the events preceding the development of lymphosarcoma. These extensive records on lymphosarcoma associated blood parameters have established that all of the inoculated animals became persistently BLV infected. However, after more than five years of incubation, no cases of lymphosarcoma developed. Consequently, during the past seven months, five of these well characterized animals have been subjected to frequent BLV re-exposure in order to study BLV-host interactions in previously infected adults and to potentially accelerate tumor formation in these animals.

Sorensen, D.K.



Ionizing radiation and risk of chronic lymphocytic leukemia in the 15-country study of nuclear industry workers.  


In contrast to other types of leukemia, chronic lymphocytic leukemia (CLL) has long been regarded as non-radiogenic, i.e. not caused by ionizing radiation. However, the justification for this view has been challenged. We therefore report on the relationship between CLL mortality and external ionizing radiation dose within the 15-country nuclear workers cohort study. The analyses included, in seven countries with CLL deaths, a total of 295,963 workers with more than 4.5 million person-years of follow-up and an average cumulative bone marrow dose of 15 mSv; there were 65 CLL deaths in this cohort. The relative risk (RR) at an occupational dose of 100 mSv compared to 0 mSv was 0.84 (95% CI 0.39, 1.48) under the assumption of a 10-year exposure lag. Analyses of longer lag periods showed little variation in the RR, but they included very small numbers of cases with relatively high doses. In conclusion, the largest nuclear workers cohort study to date finds little evidence for an association between low doses of external ionizing radiation and CLL mortality. This study had little power due to low doses, short follow-up periods, and uncertainties in CLL ascertainment from death certificates; an extended follow-up of the cohorts is merited and would ideally include incident cancer cases. PMID:18959468

Vrijheid, Martine; Cardis, Elisabeth; Ashmore, Patrick; Auvinen, Anssi; Gilbert, Ethel; Habib, Rima R; Malker, Hans; Muirhead, Colin R; Richardson, David B; Rogel, Agnes; Schubauer-Berigan, Mary; Tardy, Hélène; Telle-Lamberton, Maylis



Expression of the monocyte chemoattractant protein-1 receptor CCR2 is increased in hypercholesterolemia: differential effects of plasma lipoproteins on monocyte function  

Microsoft Academic Search

Monocytes are recruited from the circulation into the subendothelial space where they differentiate into mature macrophages and internalize modified lipoproteins to become lipid-laden foam cells. The accumulation of monocytes is mediated by the interaction of locally pro- duced chemoattractant protein-1 (MCP-1) with its receptor CCR2. The objective of the present study is to demonstrate the differential effects of plasma lipoproteins

Ki Hoon Han; Ki Ok Han; Simone R. Green; Oswald Quehenberger


The interaction of human monocytes, monocyte-derived macrophages, and polymorphonuclear neutrophils with caspofungin (MK-0991), an echinocandin, for antifungal activity against Aspergillus fumigatus  

Microsoft Academic Search

The collaboration between human effector cells and caspofungin (MK-0991), a 1,3-?-D glucan synthase inhibitor, was studied for antifungal activity against Aspergillus fumigatus. Caspofungin was co-cultured for 24h with either human monocytes (Monos), monocyte-derived macrophages (MDM), or polymorphonuclear neutrophils (PMN) against germlings of A. fumigatus and antifungal activity assessed using the XTT metabolic assay. Caspofungin at 0.1 ?g\\/ml and 0.05 ?g\\/ml

Tom Chiller; Kouros Farrokhshad; Elmer Brummer; David A Stevens



Pleiotropic effects of atorvastatin on monocytes in atherosclerotic patients.  


The objective of this study was to investigate the gene expression signature of monocyte/macrophages and the pleiotropic effects of atorvastatin on monocytes in atherosclerotic patients. Forty patients with coronary heart diseases were randomly assigned to double-blind therapy with either 20 or 80 mg per day of atorvastatin. Follow-up visits occurred at weeks 6 and 12, including complete chemistry and lipid analyses and quantification of 14 target genes in monocytes. After 12 weeks of therapy, both groups gained beneficial alterations in lipid profiles. Both groups experienced significant reductions in gene expression of lipoprotein-associated phospholipase A2, CD13, leptin receptor, matrix metalloproteases-1, legumain, and prolyl oligopeptidase after 12 weeks of therapy. Only tumor protein 53 was increased in the atorvastatin 80-mg group. Moreover, nonsignificant interactions between dosage and duration of therapy were found. The pleiotropic effects of statins in atherosclerotic patients include increased expression of genes involved in apoptosis of monocyte/macrophage, inhibition of inflammatory responses, antioxidant properties, prevention of foam cell formation, and stabilization of atherosclerotic plaques. This property fuels potential clinical significance. PMID:19808950

Wang, Zhi-hao; Liu, Xiao-lin; Zhong, Ming; Zhang, Li-ping; Shang, Yuan-yuan; Hu, Xiao-yan; Li, Li; Zhang, Yun; Deng, Jing-ti; Zhang, Wei



Differential expression of TNFR1 (CD120a) and TNFR2 (CD120b) on subpopulations of human monocytes  

PubMed Central

Background Three subpopulations of monocytes can be distinguished in human blood: classical (CD14++CD16?), intermediate (CD14++CD16+), and nonclassical (CD14+CD16++). CD16 expressing monocytes are expanded in patients with sarcoidosis and in various other inflammatory diseases. In sarcoidosis, it is unclear whether either intermediate, nonclassical or both CD16 expressing monocytes are responsible for this increase. Data relating to the monocyte subpopulations is receiving increasing attention, but the expression of TNF receptors on these subpopulations has not been studied thus far. The aim of this study was to determine frequencies of monocyte subpopulations and their expression of TNFR1 and TNFR2 in both sarcoidosis patients and healthy controls. Methods Peripheral blood cells of sarcoidosis patients and healthy controls were stained for the markers HLA-DR, CD14, CD16, CD120a and CD120b. Cells were measured on a FACSCalibur and analyzed with FlowJo. We used Student’s t-test and a parametric One-way ANOVA for statistical analysis. Results Sarcoidosis patients had a significant higher frequency of intermediate monocytes than healthy controls. Significant differences in TNF receptor expression were found between the monocyte subpopulations, both in sarcoidosis patients as well as in healthy controls: intermediates expressed more TNFR1 than classicals and nonclassicals and nonclassicals expressed more TNFR2 than intermediates, whereas intermediates showed higher expression than classicals. Conclusions In both sarcoidosis patients and healthy controls intermediate monocytes show the highest expression level of TNFR1 among monocyte subpopulations and nonclassical monocytes show the highest expression level of TNFR2. These findings, as wells as the higher frequency of intermediate monocytes in sarcoidosis patients, provide evidence for the existence of two functionally-distinct CD16 expressing monocyte subpopulations.



Functional and morphologic characteristics of the leukemic cells of a patient with acute monocytic leukemia: correlation with clinical features.  


The clinical course of a patient with acute monocytic leukemia and prominent infiltration of the skin and testes is described. In vitro studies demonstrated that the circulating monocyte precursors were capable of adherence to nylon fibers, and phagocytosis of bacteria and latex particles. In vivo, migration of leukemic cells to skin windows was observed. Extreme nuclear folding, marked surface activity, and morphologic features suggesting nuclear and cytoplasmic maturation were seen by light and electron microscopy. The presence of morphologically and functionally more differentiated monocytic cells may account for the marked tiuuse invasion in this patient and, possibly, in other patients with monocytic leukemia. PMID:1055611

Schiffer, C A; Sanel, F T; Stechmiller, B K; Wiernik, P H



Generation of Mature Monocyte-Derived Dendritic Cells in the Presence of Heparin and Monocyte Conditioned Medium: Phenotypic and Functional Comparison  

PubMed Central

Background: Dendritic cells (DC) induce tumor or pathogen-specific T cell responses in humans. Several laboratories have developed culture systems, including maturation factors for human DC from peripheral blood monocytes. We comprehensively compared standard maturation stimulus, an autologous monocyte-conditioned medium (MCM), with heparin for their ability to promote uniformly mature DC that elicit T cell responses. Methods: A short (4-day) priming of plastic adherent monocytes with granulocyte-macrophage colony stimulating factor (GM-CSF) and IL-4 with or without heparin was followed by 48-hour incubation in MCM to generate fully mature and stable DC. Phenotypic and functional analyses were carried out using anti-CD14 and anti-CD83 monoclonal antibodies, and mixed lymphocyte reaction, respectively. Results: We found that fully matured DC with a large amount of cytoplasm and copious dendritic projections were visible at the end of culturing period in the presence of MCM, heparin and MCM plus heparin. Thus, DC generated with these maturation factors are non-adherent and have typical satellite morphology. Flow cytometric analysis using anti-CD14 (monocyte marker) and anti-CD83 (mature DC marker) revealed that expression of CD14 decreased in MCM plus heparin-treated DC, and the expression of CD83 was increased when heparin and MCM used as a maturation factor. Functionally, MCM and MCM plus heparin-treated DC showed stronger mixed leukocyte reaction than heparin alone. Conclusion: These results support the use of the MCM with heparin as maturation factor that could result in functionally mature monocyte-derived DC in comparison to either MCM or heparin alone.

Delirezh, Nowruz; Majedi, Leila; Asri Rezaei, Siamak; Ranjkeshzadeh, Hadi



Myxoma viral serpin, Serp-1, inhibits human monocyte adhesion through regulation of actin-binding protein filamin B.  


Serp-1 is a secreted myxoma viral serine protease inhibitor (serpin) with proven, highly effective, anti-inflammatory defensive activity during host cell infection, as well as potent immunomodulatory activity in a wide range of animal disease models. Serp-1 binds urokinase-type plasminogen activator (uPA) and the tissue-type PA, plasmin, and factor Xa, requiring uPA receptor (uPAR) for anti-inflammatory activity. To define Serp-1-mediated effects on inflammatory cell activation, we examined the association of Serp-1 with monocytes and T cells, effects on cellular migration, and the role of uPAR-linked integrins and actin-binding proteins in Serp-1 cellular responses. Our results show that Serp-1 associates directly with activated monocytes and T lymphocytes, in part through interaction with uPAR (P<0.001). Serp-1, but not mammalian serpin PA inhibitor-1 (PAI-1), attenuated cellular adhesion to the extracellular matrix. Serp-1 and PAI-1 reduced human monocyte and T cell adhesion (P<0.001) and migration across endothelial monolayers in vitro (P<0.001) and into mouse ascites in vivo (P<0.001). Serp-1 and an inactive Serp-1 mutant Serp-1(SAA) bound equally to human monocytes and T cells, but a highly proinflammatory mutant, Serp-1(Ala(6)), bound less well to monocytes. Serp-1 treatment of monocytes increased expression of filamin B actin-binding protein and reduced CD18 (beta-integrin) expression (P<0.001) in a uPAR-dependent response. Filamin colocalized and co-immunoprecipitated with uPAR, and short interference RNA knock-down of filamin blocked Serp-1 inhibition of monocyte adhesion. We report here that the highly potent, anti-inflammatory activity of Serp-1 is mediated through modification of uPAR-linked beta-integrin and filamin in monocytes, identifying this interaction as a central regulatory axis for inflammation. PMID:19052145

Viswanathan, Kasinath; Richardson, Jakob; Togonu-Bickersteth, Babajide; Dai, Erbin; Liu, Liying; Vatsya, Pracha; Sun, Yun-ming; Yu, Jeff; Munuswamy-Ramanujam, Ganesh; Baker, Henry; Lucas, Alexandra R



Lymphocyte infiltration and activation in iris-ciliary body and anterior chamber of mice in corneal allograft rejection  

PubMed Central

AIM To investigate the infiltration and activation of lymphocyte in iris-ciliary body and anterior chamber after allogenic penetrating keratoplasty (PK), for further revealing the role of iris-ciliary body in corneal allograft immune rejection. METHODS In the mice models of PK, BALB/C mice received orthotopic isografts (n =35) or C57BL/6 donor allografts (n =25). Grafts were examined daily for 3 weeks by slit-lamp microscopy and scored for opacity. The infiltration of CD4+ T lymphocyte in iris-ciliary body and anterior chamber was examined by immunohistology and the mRNA of CD80 and CD86 in both cornea graft and iris-ciliary body by RT-PCR was analyzed in allograft recipient at days 3, 6, 10 and the day when graft rejection occurred. Isograft recipients were examined as control at the corresponding time points. Transmission electron microscope was used to study the ultrastructure, especially cell infiltration, of iris-cilary body and corneal graft at day 3, 7 and the day when rejection occurred after allogenic PK. RESULTS Rejection was observed in all the allograft recipients followed more than 10 days, at a median time of 15 days (range 12-18 days), but not in any of isografts. CD4+ T cells were first detected at day 6 after transplantation in limbus and Ciliary body, and then in the stroma of recipient, iris, anterior chamber and corneal allograft with an increased number until graft rejection occurred. CD80 and CD86 mRNA were detected under RT-PCR examination in both graft and iris-ciliary body of allograft recipient, but not in any of isograft recipient. Three days after operation, lymphocytes and monocytes macrophages were visible in iris blood vessels and the anterior chamber, and vascular endothelial cell proliferation and activation were significant under transmission electron microscopy examination. At day 7, corneal endothelial cells became thinner. Lymphocytes and mononuclear macrophages were found with great number in the anterior chamber and adhered to the corneal endothelium. Blood vessels in iris increased and were filled with lymphocytes. And lymphocytes were detected to migrate through endothelial cell gap out of vessels. When allograft rejection occurred, macrophages attached to endothelial cells with large number of lymphocytes and macrophages infiltrating in iris. CONCLUSION Lymphocyte infiltration and activation occurred in iris-ciliary body after allogenic PK, and the lymphocytes could migrate from iris blood vessel to the anterior chamber, which might play an important role in corneal allograft immune rejection.

Wang, Fu-Hua; Chen, Min; Liu, Ting; Zang, Xin-Jie; Gong, Hua-Qing; Shi, Wei-Yun



Chemokine receptor Ccr2 is critical for monocyte accumulation and survival in West Nile virus encephalitis.  


West Nile virus (WNV) is a re-emerging pathogen responsible for outbreaks of fatal meningoencephalitis in humans. Previous studies have suggested a protective role for monocytes in a mouse model of WNV infection, but the molecular mechanisms have remained unclear. In this study, we show that genetic deficiency in Ccr2, a chemokine receptor on Ly6c(hi) inflammatory monocytes and other leukocyte subtypes, markedly increases mortality due to WNV encephalitis in C57BL/6 mice; this was associated with a large and selective reduction of Ly6c(hi) monocyte accumulation in the brain. WNV infection in Ccr2(+/+) mice induced a strong and highly selective monocytosis in peripheral blood that was absent in Ccr2(-/-) mice, which in contrast showed sustained monocytopenia. When a 1:1 mixture of Ccr2(+/+) and Ccr2(-/-) donor monocytes was transferred by vein into WNV-infected Ccr2(-/-) recipient mice, monocyte accumulation in the CNS was not skewed toward either component of the mixture, indicating that Ccr2 is not required for trafficking of monocytes from blood to brain. We conclude that Ccr2 mediates highly selective peripheral blood monocytosis during WNV infection of mice and that this is critical for accumulation of monocytes in the brain. PMID:21131425

Lim, Jean K; Obara, Christopher J; Rivollier, Aymeric; Pletnev, Alexander G; Kelsall, Brian L; Murphy, Philip M



Role of exercise intensities in oxidized low-density lipoprotein-mediated redox status of monocyte in men.  


Exercise significantly influences the progression of atherosclerosis. Oxidized LDL (ox-LDL), as a stimulator of oxidative stress, facilitates monocyte-related atherogenesis. This study investigates how exercise intensity impacts ox-LDL-mediated redox status of monocytes. Twenty-five sedentary healthy men exercised mildly, moderately, and heavily (i.e., 40, 60, and 80% maximal oxygen consumption, respectively) on a bicycle ergometer. Reactive oxygen species (ROS) production, cytosolic and mitochondrial superoxide dismutase (c-SOD and m-SOD, respectively) activities, and total and reduced-form gamma-glutamylcysteinyl glycine (t-GSH and r-GSH, respectively) contents in monocytes mediated by ox-LDL were measured. This experiment obtained the following findings: 1) ox-LDL increased monocyte ROS production and was accompanied by decreased c-SOD and m-SOD activities, as well as t-GSH and r-GSH contents, whereas treating monocytes with diphenyleneiodonium (DPI) (a NADPH oxidase inhibitor) or rotenone/2-thenoyltrifluoroacetone (TTFA) (mitochondrial complex I/II inhibitors) hindered ox-LDL-induced monocyte ROS production; 2) production of ROS and reduction of m-SOD activity and r-GSH content in monocyte by ox-LDL were enhanced by heavy exercise and depressed by mild and moderate exercise; and 3) heavy exercise augmented the inhibition of ox-LDL-induced monocyte ROS production by DPI and rotenone/TTFA, whereas these DPI- and rotenone/TTFA-mediated monocyte ROS productions were unchanged in response to mild and moderate exercise. We conclude that heavy exercise increases ox-LDL-induced monocyte ROS production, possibly by decreasing m-SOD activity and r-GSH content in monocytes. However, mild and moderate exercise likely protects individuals against suppression of anti-oxidative capacity of monocyte by ox-LDL. PMID:16728523

Wang, Jong-Shyan; Lee, Tan; Chow, Shu-Er



The generation of NGF-secreting primary rat monocytes: A comparison of different transfer methods  

PubMed Central

Nerve growth factor (NGF), a member of the neurotrophin family, is responsible for the maintenance and survival of cholinergic neurons in the basal forebrain. The degeneration of cholinergic neurons and reduced acetycholine levels are hallmarks of Alzheimer's disease (AD) as well as associated with learning and memory deficits. Thus far, NGF has proven the most potent neuroprotective molecule against cholinergic neurodegeneration. However, delivery of this factor into the brain remains difficult. Recent studies have begun to elucidate the potential use of monocytes as vehicles for therapeutic delivery into the brain. In this study, we employed different transfection and transduction methods to generate NGF-secreting primary rat monocytes. Specifically, we compared five methods for generating NGF-secreting monocytes: (1) cationic lipid-mediated transfection (Effectene and FuGene), (2) classical electroporation, (3) nucleofection, (4) protein delivery (Bioporter) and (5) lentiviral vectors. Here, we report that classical transfection methods (lipid-mediated transfection, electroporation, nucleofection) are inefficient tools for proper gene transfer into primary rat monocytes. We demonstrate that lentiviral infection and Bioporter can successfully transduce/load primary rat monocytes and produce effective NGF secretion. Furthermore, our results indicate that NGF is bioactive and that Bioporter-loaded monocytes do not appear to exhibit any functional disruptions (i.e. in their ability to differentiate and phagocytose beta-amyloid). Taken together, our results show that primary monocytes can be effectively loaded or transduced with NGF and provides information on the most effective method for generating NGF-secreting primary rat monocytes. This study also provides a basis for further development of primary monocytes as therapeutic delivery vehicles to the diseased AD brain.

Hohsfield, Lindsay A.; Geley, Stephan; Reindl, Markus; Humpel, Christian



Acute Lymphocytic Leukemia  


... may be reprinted for personal, noncommercial use only. Acute lymphocytic leukemia By Mayo Clinic staff Original Article: Definition Symptoms Causes Risk factors Preparing for ...


Acute Lymphocytic Leukemia  


... a third party. HPF: SEER Stat Fact Sheets: Acute Lymphocytic Leukemia Cancer: It is estimated that 6,070 men ... 1,430 men and women will die of acute lymphocytic leukemia in 2013 1 X Close Table I-1 ( ...


Acute Lymphocytic Leukemia  


... hard for blood to do its work. In acute lymphocytic leukemia (ALL), also called acute lymphoblastic leukemia, there are too ... of white blood cells called lymphocytes or lymphoblasts. ALL is the most common type of cancer in ...


Longitudinal study of patients with idiopathic isolated TSH deficiency: possible progression of pituitary dysfunction in lymphocytic adenohypophysitis.  


The relationship between isolated TSH deficiency and hypophysitis was studied. Six patients (five women and one man) with idiopathic isolated TSH deficiency were longitudinally investigated with an interval of 31 to 60 months. Clinical symptoms, laboratory results and endocrine function were investigated as well as pituitary magnetic resonance imaging (MRI) at the start and the end of the study. Clinically, initial symptoms due to hypothyroidism were ameliorated by the thyroid hormone replacement in all patients. Oligomenorrhea newly appeared during the study in three patients, although no other symptoms appeared. Serum fT3 and fT4 levels were within the reference ranges, and serum TSH level and its response to TRH stimulation remained low in all patients. Peak plasma GH level during GRH stimulation was significantly (p<0.03) decreased, at the end of the study as compared with the start. Peak plasma FSH level to LHRH stimulation was significantly (p<0.03) decreased as well as basal FSH level. In contrast, peak of prolactin during TRH stimulation was significantly (p<0.03) increased at the end of the study as compared with the start as well as basal prolactin level. Endocrine features at the end of the study were compatible with those of lymphocytic adenohypophysitis (LAH). MRI of the pituitary gland showed empty sella in one patient and slight swelling in two patients. These findings remained unchanged during the study period. One patient underwent pituitary biopsy, with histological examination showing atypical form of LAH. LAH can cause idiopathic isolated TSH deficiency and can functionally progress to combine dysfunction of the pituitary gland. PMID:16896268

Hashimoto, Ken-Ichi; Yamakita, Noriyoshi; Ikeda, Tsuneko; Matsuhisa, Takashi; Kuwayama, Akio; Sano, Toshiaki; Hashimoto, Kozo; Yasuda, Keigo



Volume regulation by human lymphocytes. Identification of differences between the two major lymphocyte subpopulations.  

PubMed Central

Following exposure to hypotonic media, human peripheral blood lymphocytes swell initially but restore their isotonic volume within minutes. In contrast, tonsillar lymphocytes demonstrate a similar initial phase of swelling but fail to restore their isotonic volume. We have studied the ionic basis for this second or regulatory volume decrease (RVD) phase using lymphocytes from peripheral blood, tonsil, and thymus. RVD was characterized by 86Rb efflux and a decrease in K+ content. The increase in K+ permeability in response to hypotonic challenge was characteristic for T lymphocytes obtained from peripheral blood, tonsil, or thymus. B lymphocytes showed only a modest increase in K+ permeability and consequently little RVD. The data confirm that the response of peripheral blood and tonsillar lymphocytes to a hypotonic environment can be accounted for by differences in the proportions of T and B cells, and the differential behaviour of B and T lymphocytes is based on differences in membrane permeability to K+ upon swelling.

Cheung, R K; Grinstein, S; Gelfand, E W



Sphingosine 1-phosphate interferes on the differentiation of human monocytes into competent dendritic cells.  


Sphingosine 1-phosphate (S1P) is a lipidic messenger known to exert several physiological functions within the cell. We tested here whether the stimulation of human monocytes with different doses of S1P might interfere with their differentiation into competent dendritic cells (DC). Monocytes cultured with granulocyte macrophage colony stimulating factor, interleukin-4 (IL-4) and S1P differentiated into a DC population lacking CD1a molecules on the surface and acquired some aspects of mature DC (mDC), though in the absence of maturation stimuli. When stimulated with lipopolisaccharide (LPS), CD1a(-) DC produce high amounts of tumour necrosis factor-alpha and IL-10, but not IL-12. Accordingly, these CD1a(-) DC were not capable of stimulating allogenic T lymphocytes so well as CD1a(+) DC generated from untreated monocytes and maturated with LPS. S1P monocyte-derived DC lost their polarizing capacity abrogating the production of gamma-interferon/IL-4 by co-cultured naïve CD4(+)CD45RA(+) T cells. Our findings suggest a mechanism through which S1P can favour the development of immune-related pathological states. PMID:17212771

Martino, A; Volpe, E; Auricchio, G; Izzi, V; Poccia, F; Mariani, F; Colizzi, V; Baldini, P M



Functional capabilities of marmoset T and B lymphocytes in primary in vitro antibody formation  

SciTech Connect

In vitro tests of T- and B-lymphocyte function of two marmoset species, Saguinus fuscicollis and Saguinus oedipus, were examined to explore the lower immune response profile previously reported for S. o. oedipus. Experiments with trinitrophenyl-lipopolysaccharide (TNP-LPS) revealed peripheral blood leukocytes (PBL) from both species capable of antibody formation. This response was both T cell and monocyte independent; indeed, removal of T cells led to an enhanced response, indicating a regulatory role for this cell in each species. Studies with the nonmitogenic form of TNP-LPS, trinitrophenyl-base-hydrolyzed-lipopolysaccharide, revealed that plaque-forming cells could be obtained from S. fuscicollis PBL while S. o. oedipus PBL were unresponsive. This report also demonstrates that hemopoietic chimerism, a feature common to all marmosets, has a negative influence on antibody-forming capabilities.

Nickerson, D.A.; Gengozian, N.



Benefit from B-Lymphocyte Depletion Using the Anti-CD20 Antibody Rituximab in Chronic Fatigue Syndrome. A Double-Blind and Placebo-Controlled Study  

Microsoft Academic Search

BackgroundChronic fatigue syndrome (CFS) is a disease of unknown aetiology. Major CFS symptom relief during cancer chemotherapy in a patient with synchronous CFS and lymphoma spurred a pilot study of B-lymphocyte depletion using the anti-CD20 antibody Rituximab, which demonstrated significant clinical response in three CFS patients.Methods and FindingsIn this double-blind, placebo-controlled phase II study (NCT00848692), 30 CFS patients were randomised

Øystein Fluge; Ove Bruland; Kristin Risa; Anette Storstein; Einar K. Kristoffersen; Dipak Sapkota; Halvor Næss; Olav Dahl; Harald Nyland; Olav Mella; Markus Reindl



The Blood Neutrophil to Lymphocyte Ratio Correlates with Clinical Status in Children with Cystic Fibrosis: A Retrospective Study  

PubMed Central

Purpose The blood neutrophil to lymphocyte ratio (NLR) has been identified as a potentially useful marker of clinical outcome in disease states with an inflammatory component. The objective of this study was to evaluate the relationship between NLR and clinical status in children with cystic fibrosis. Methods This was a retrospective chart review. Data collected included NLR, body mass index, and forced expiratory volume in 1 second (FEV1) while asymptomatic, and during hospitalizations for pulmonary exacerbation. An NLR breakpoint of 3 was used for comparisons of body mass index and FEV1. Results A total of 159 charts were reviewed. An NLR ? 3 was significantly associated with lower body mass index and lower FEV1. NLR during hospitalization was significantly higher than NLR while asymptomatic. NLR measured during the first 3 months of life was negatively correlated with FEV1 at age 12. Conclusion NLR correlates with clinical status in children with cystic fibrosis and may be a useful biomarker in this population.

O'Brien, Catherine E.; Price, Elvin T.



Monocyte cell surface glycosaminoglycans positively modulate IL-4-induced differentiation toward dendritic cells.  


IL-4 induces the differentiation of monocytes toward dendritic cells (DCs). The activity of many cytokines is modulated by glycosaminoglycans (GAGs). In this study, we explored the effect of GAGs on the IL-4-induced differentiation of monocytes toward DCs. IL-4 dose-dependently up-regulated the expression of DC-specific ICAM-3-grabbing nonintegrin (DC-SIGN), CD80, CD206, and CD1a. Monocytes stained positive with Abs against heparan sulfate (HS) and chondroitin sulfate (CS) B (CSB; dermatan sulfate), but not with Abs that recognize CSA, CSC, and CSE. Inhibition of sulfation of monocyte/DC cell surface GAGs by sodium chlorate reduced the reactivity of sulfate-recognizing single-chain Abs. This correlated with hampered IL-4-induced DC differentiation as evidenced by lower expression of DC-SIGN and CD1a and a decreased DC-induced PBL proliferation, suggesting that sulfated monocyte cell surface GAGs support IL-4 activity. Furthermore, removal of cell surface chondroitin sulfates by chondroitinase ABC strongly impaired IL-4-induced STAT6 phosphorylation, whereas removal of HS by heparinase III had only a weak inhibitory effect. IL-4 bound to heparin and CSB, but not to HS, CSA, CSC, CSD, and CSE. Binding of IL-4 required iduronic acid, an N-sulfate group (heparin) and specific O sulfates (CSB and heparin). Together, these data demonstrate that monocyte cell surface chondroitin sulfates play an important role in the IL-4-driven differentiation of monocytes into DCs. PMID:18322173

den Dekker, Els; Grefte, Sander; Huijs, Tonnie; ten Dam, Gerdy B; Versteeg, Elly M M; van den Berk, Lieke C J; Bladergroen, Bellinda A; van Kuppevelt, Toin H; Figdor, Carl G; Torensma, Ruurd



RhoA is required for monocyte tail retraction during transendothelial migration  

PubMed Central

Transendothelial migration of monocytes is the process by which monocytes leave the circulatory system and extravasate through the endothelial lining of the blood vessel wall and enter the underlying tissue. Transmigration requires coordination of alterations in cell shape and adhesive properties that are mediated by cytoskeletal dynamics. We have analyzed the function of RhoA in the cytoskeletal reorganizations that occur during transmigration. By loading monocytes with C3, an inhibitor of RhoA, we found that RhoA was required for transendothelial migration. We then examined individual steps of transmigration to explore the requirement for RhoA in extravasation. Our studies showed that RhoA was not required for monocyte attachment to the endothelium nor subsequent spreading of the monocyte on the endothelial surface. Time-lapse video microscopy analysis revealed that C3-loaded monocytes also had significant forward crawling movement on the endothelial monolayer and were able to invade between neighboring endothelial cells. However, RhoA was required to retract the tail of the migrating monocyte and complete diapedesis. We also demonstrate that p160ROCK, a serine/threonine kinase effector of RhoA, is both necessary and sufficient for RhoA-mediated tail retraction. Finally, we find that p160ROCK signaling negatively regulates integrin adhesions and that inhibition of RhoA results in an accumulation of ?2 integrin in the unretracted tails.

Worthylake, Rebecca A.; Lemoine, Sean; Watson, Joanna M.; Burridge, Keith



Chemokine Receptor CCR2 is Critical for Monocyte Accumulation and Survival in West Nile Virus Encephalitis  

PubMed Central

West Nile virus (WNV) is a re-emerging pathogen responsible for outbreaks of fatal meningoencephalitis in humans. Previous studies have suggested a protective role for monocytes in a mouse model of WNV infection, but the molecular mechanisms have remained unclear. Here we show that genetic deficiency in Ccr2, a chemokine receptor on Ly6chi inflammatory monocytes and other leukocyte subtypes, markedly increases mortality due to WNV encephalitis in C57Bl/6 mice; this was associated with a large and selective reduction of Ly6chi monocyte accumulation in the brain. WNV infection in Ccr2+/+ mice induced a strong and highly selective monocytosis in peripheral blood that was absent in Ccr2?/? mice, which in contrast showed sustained monocytopenia. When a 1:1 mixture of Ccr2+/+ and Ccr2?/? donor monocytes was transferred by vein into WNV-infected Ccr2?/? recipient mice, monocyte accumulation in the CNS was not skewed toward either component of the mixture, indicating that Ccr2 is not required for trafficking of monocytes from blood to brain. We conclude that Ccr2 mediates highly selective peripheral blood monocytosis during WNV infection of mice and that this is critical for accumulation of monocytes in the brain.

Lim, Jean K.; Obara, Christopher J.; Rivollier, Aymeric; Pletnev, Alexander G.; Kelsall, Brian L.; Murphy, Philip M.



Effect of Zinc and Nitric Oxide on Monocyte Adhesion to Endothelial Cells under Shear Stress  

PubMed Central

This study describes the effect of zinc on monocyte adhesion to endothelial cells under different shear stress regimens, which may trigger atherogenesis. Human umbilical vein endothelial cells were exposed to steady shear stress (15 dynes/cm2 or 1 dyne/cm2) or reversing shear stress (time average 1 dyne/cm2) for 24 hours. In all shear stress regimes, zinc deficiency enhanced THP-1 cell adhesion, while heparinase III reduced monocyte adhesion following reversing shear stress exposure. Unlike other shear stress regimes, reversing shear stress alone enhanced monocyte adhesion, which may be associated with increased H2O2 and superoxide together with relatively low levels of nitric oxide (NO) production. L-NG-Nitroarginine methyl ester (L-NAME) treatment increased monocyte adhesion under 15 dynes/cm2 and under reversing shear stress. After reversing shear stress monocyte adhesion dramatically increased with heparinase III treatment followed by a zinc scavenger. Static culture experiments supported the reduction of monocyte adhesion by zinc following endothelial cell cytokine activation. These results suggest that endothelial cell zinc levels are important for the inhibition of monocyte adhesion to endothelial cells, and may be one of the key factors in the early stages of atherogenesis.

Lee, Sungmun; Eskin, Suzanne G.; Shah, Ankit K.; Schildmeyer, Lisa A.; McIntire, Larry V.



On the prediction of monocyte deposition in abdominal aortic aneurysms using computational fluid dynamics.  


In abdominal aortic aneurysm disease, the aortic wall is exposed to intense biological activity involving inflammation and matrix metalloproteinase-mediated degradation of the extracellular matrix. These processes are orchestrated by monocytes and rather than affecting the aorta uniformly, damage and weaken focal areas of the wall leaving it vulnerable to rupture. This study attempts to model numerically the deposition of monocytes using large eddy simulation, discrete phase modelling and near-wall particle residence time. The model was first applied to idealised aneurysms and then to three patient-specific lumen geometries using three-component inlet velocities derived from phase-contrast magnetic resonance imaging. The use of a novel, variable wall shear stress-limiter based on previous experimental data significantly improved the results. Simulations identified a critical diameter (1.8 times the inlet diameter) beyond which significant monocyte deposition is expected to occur. Monocyte adhesion occurred proximally in smaller abdominal aortic aneurysms and distally as the sac expands. The near-wall particle residence time observed in each of the patient-specific models was markedly different. Discrete hotspots of monocyte residence time were detected, suggesting that the monocyte infiltration responsible for the breakdown of the abdominal aortic aneurysm wall occurs heterogeneously. Peak monocyte residence time was found to increase with aneurysm sac size. Further work addressing certain limitations is needed in a larger cohort to determine clinical significance. PMID:23886969

Hardman, David; Doyle, Barry J; Semple, Scott Ik; Richards, Jennifer Mj; Newby, David E; Easson, William J; Hoskins, Peter R



Ursolic Acid Protects Diabetic Mice Against Monocyte Dysfunction and Accelerated Atherosclerosis  

PubMed Central

Aims Accelerated atherosclerosis is a major diabetic complication initiated by the enhanced recruitment of monocytes into the vasculature. In this study, we examined the therapeutic potential of the phytonutrients ursolic acid (UA) and resveratrol (RES) in preventing monocyte recruitment and accelerated atherosclerosis. Methods and Results Dietary supplementation with either RES or UA (0.2%) protected against accelerated atherosclerosis induced by streptozotocin in high-fat diet-fed LDL receptor-deficient mice. However, mice that received dietary UA for 11 weeks were significantly better protected and showed a 53% reduction in lesion formation while mice fed a RES-supplemented diet showed only a 31% reduction in lesion size. Importantly, UA was also significantly more effective in preventing the appearance of proinflammatory GR-1high monocytes induced by these diabetic conditions and reducing monocyte recruitment into MCP-1-loaded Matrigel plugs implanted into these diabetic mice. Oxidatively-stressed THP-1 monocytes mimicked the behavior of blood monocytes in diabetic mice and showed enhanced responsiveness to monocyte chemoattractant protein-1 (MCP-1) without changing MCP-1 receptor (CCR2) surface expression. Pretreatment of THP-1 monocytes with RES or UA (0.3 – 10 ?M) for 15 h resulted in the dose-dependent inhibition of H2O2-accelerated chemotaxis in response to MCP-1, but with an IC50 of 0.4 ?M, UA was 2.7-fold more potent than RES. Conclusion Dietary UA is a potent inhibitor of monocyte dysfunction and accelerated atherosclerosis induced by diabetes. These studies identify ursolic acid as a potential therapeutic agent for the treatment of diabetic complications, including accelerated atherosclerosis, and provide a novel mechanism for the anti-atherogenic properties of ursolic acid.

Ullevig, Sarah L.; Zhao, Qingwei; Zamora, Debora; Asmis, Reto



CD2 activation of human lamina propria lymphocytes reduces CD3 responsiveness  

PubMed Central

Lamina propria lymphocytes (LPLs) are thought to be antigen-activated memory T cells. Yet, they respond better to ligation of the CD2 receptor than the CD3 receptor by mitogenic antibodies. This study defines their constitutive state of activation and relates it to their CD3 hyporesponsiveness. The activated state of LPLs was demonstrated by their heightened display of the activated CD2 epitope, T113. Constitutive CD2 activation was shown by the reduction in spontaneous proliferation when the CD2–CD58 interaction was blocked. LPLs preferentially recognized CD58 rather than the major histocompatibility complex molecules on monocytes, triggering proliferation and interferon-? (IFN-?) secretion that was inhibited by blocking the CD2–CD58 interaction. To determine whether CD2 activation of LPLs contributes to their CD3 hyporesponsiveness, they were first stimulated with mitogenic CD2 antibodies and then tested for CD3-induced proliferation. The responses were greatly reduced by prior CD2 stimulation compared with LPLs cultured in medium alone. This effect was not caused by apoptosis or by changes in CD3 expression induced by CD2 triggering. This study shows that LPLs are constitutively activated through CD2, that they preferentially recognize CD58 on monocytes and that CD2 stimulation leads to CD3 hyporesponsiveness.

Ebert, Ellen C



Interaction of alphaviruses with human peripheral leukocytes: in vitro replication of Venezuelan equine encephalomyelitis virus in monocyte cultures.  

PubMed Central

Human peripheral blood leukocytes (PBL) were examined for their ability to support growth of several group A arboviruses in vitro. Cells were refractory to infection with eastern (EEE) and western (WEE) equine encephalitis viruses, whereas Venezuelan equine encephalomyelitis (VEE) virus was shown to infect and replicate to a substantially high titer. When PBL were fractionated into purified subpopulations, only the monocytes were susceptible to predictive VEE virus infection. Lymphocytes treated 24 h before virus inoculation with phytohemagglutinin (10 microgram/ml) were capable of propagating significant amounts of VEE virus. A monocytic cell line, J-111, was also susceptible to infection with VEE, EEE, and WEE viruses, whereas a lymphocytic cell line, Raji, was refractory. Additional information on the participation of PBL during human infection with these viruses may add considerably to our understanding of their differing pathogenicities and clinical pictures.

Levitt, N H; Miller, H V; Edelman, R



Lymphocyte-Platelet Crosstalk in Graves' Disease.  


OBJECTIVE:: Platelets can modulate lymphocytes' role in the pathophysiology of thyroid autoimmune diseases. The present study was performed to clarify the status of platelet-lymphocyte subpopulations aggregation in circulating blood in patients with Graves' disease (GD). METHODS:: One hundred and fifty patients with GD (GD group) and 45 hyperthyroid patients with toxic multinodular goiter (TMG group) were recruited in the study. Control group consisted 150 healthy subjects. Immunophenotyping of lymphocytes was performed by flow cytometry. Detection of lymphocyte-platelet aggregates (LPAs) was done using light microscope after Ficoll-gradient centrifugation. RESULTS:: The group of GD patients exhibited reduced CD8 lymphocyte and higher CD19 cell counts compared with TMG group and healthy controls. A greater number of activated CD3, HLA-DR+ lymphocytes were observed in GD than in TMG group and control group. GD group was characterized by lower blood platelet count (232 ± 89 × 10 cells/µL) than TMG group (251 ± 97 × 10 cells/µL; P < 0.05) and control group (262 ± 95 × 10 cells/µL; P < 0.05). In GD group, more platelet-bound lymphocytes (332 ± 91 /µL) were found than that in TMG group (116 ± 67/µL, P < 0.005) and control group (104 ± 58 /µL; P < 0.001). CONCLUSIONS:: GD is associated with higher levels of activated lymphocytes and lymphocyte-platelet aggregates. PMID:23571468

Kuznik, Boris I; Vitkovsky, Yuri A; Gvozdeva, Olga V; Solpov, Alexey V; Magen, Eli




Microsoft Academic Search

Loa C. C., T. L. Lin, C. C. W u: Factors Affecting Mitogenic Response of Turkey Lymphocytes. Acta Vet. Brno 2001, 70: 433-442. The objective of the present study was to determine the parameters and conditions for measuring mitogenic response of turkey whole blood and spleen lymphocytes that may be useful in studying cellular immunity of turkeys. Heparinized whole blood

C. C. LOA; T. L. LIN; C. C. WU



Sympathetic Nervous System and Lymphocyte Proliferation in the Fischer 344 Rat Spleen: A Longitudinal Study  

Microsoft Academic Search

Aging is associated with reduced cellular immunity, which leads to increased rates of infectious disease, cancer and autoimmunity in the elderly. Previous findings from our laboratory revealed an age-related decline in sympathetic innervation of immune organs that affects immunity. These studies suggested potential sympathetic nervous system involvement in age-induced immune dysregulation. Objectives: The purpose of this study was to longitudinally

Denise L. Bellinger; Dorian Silva; Ashley Brooke Millar; Christine Molinaro; Mark Ghamsary; Jeff Carter; Sam Perez; Dianne Lorton; Cheri Lubahn; Gerson Araujoa; Srinivasan Thyagarajan



Fractalkine Preferentially Mediates Arrest and Migration of CD16 ? Monocytes  

Microsoft Academic Search

CD16 ? monocytes represent 5-10% of peripheral blood monocytes in normal individuals and are dramatically expanded in several pathological conditions including sepsis, human immuno- deficiency virus 1 infection, and cancer. CD16 ? monocytes produce high levels of proin- flammatory cytokines and may represent dendritic cell precursors in vivo. The mechanisms that mediate the recruitment of CD16 ? monocytes into tissues

Petronela Ancuta; Ravi Rao; Ashlee Moses; Andrew Mehle; Sunil K. Shaw; F. William Luscinskas; Dana Gabuzda



Modulation of the expression of chondroitin sulfate proteoglycan in stimulated human monocytes.  


Proteoglycan biosynthesis was studied in human monocytes and monocyte-derived macrophages (MDM) after exposure to typical activators of the monocyte/macrophage system: interferon-gamma (IFN-gamma), lipopolysaccharide (LPS), and phorbol 12-myristate 13-acetate (PMA). By morphological examination, both monocytes and MDM were stimulated by these activators. Treatment with IFN-gamma resulted in a slight decrease in the expression of [35S]chondroitin sulfate proteoglycan (CSPG) in both monocytes and MDM, whereas LPS treatment increased the [35S]CSPG expression 1.8 and 2.2 times, respectively. PMA, in contrast, decreased the CSPG expression 0.4 times in monocytes, whereas MDM were stimulated to increase the biosynthesis 1.9 times. An increase in the sulfate density of the chondroitin sulfate chains was evident following differentiation of monocytes into MDM due to the expression of disulfated disaccharide units of the chondroitin sulfate E type (CS-E). However, monocytes exposed to PMA did also express disaccharides of the chondroitin sulfate E type. Furthermore, the expression of CS-E in MDM was increased 2 times following PMA treatment. An inactive phorbol ester, phorbol 12,13-diacetate, did not affect the expression of CS-E in either monocytes or MDM when compared with control cultures, suggesting that protein kinase C-dependent signal pathways may be involved in the regulation of sulfation of CSPG. Exposure to LPS or IFN-gamma did not lead to any changes in the sulfation of the chondroitin sulfate chains. PMID:2768247

Uhlin-Hansen, L; Eskeland, T; Kolset, S O



Analysis using canine peripheral blood for establishing in vitro conditions for monocyte differentiation into macrophages for Leishmania chagasi infection and T-cell subset purification.  


Canine visceral leishmaniasis (CVL) is a parasitic disease endemic in many countries, and dogs present as the major natural reservoir of the parasite, Leishmania chagasi (syn. L. infantum). Biomarkers in the canine immune system is an important technique in the course of developing vaccines and treatment strategies against CVL. New methodologies for studying the immune response of dogs during Leishmania infection and after receiving vaccines and treatments against CVL would be useful. In this context, we used peripheral blood mononuclear cells (PBMCs) from healthy dogs to evaluate procedures related to (i) establishment of in vitro conditions of monocytes differentiated into macrophages infected with L. chagasi and (ii) purification procedures of T-cell subsets (CD4(+) and CD8(+)) using microbeads. Our data demonstrated that after 5 days of differentiation, macrophages were able to induce significant phagocytic and microbicidal activity after L. chagasi infection and also showed increased frequency of parasitism and a higher parasite load. Although N-acetyl-?-d-glucosaminidase (NAG) levels presented similar levels of macrophage culture and L. chagasi infection, a progressive decrease in myeloperoxidase (MPO) levels was a hallmark over 5 days of culture. High purity levels (>90%) of CD4 and CD8 T cells were obtained on a magnetic separation column. We concluded that monocytes differentiated into macrophages at 5 days and displayed an intermediate frequency of parasitism and parasite load 72h after L. chagasi infection. Furthermore, the purification system using canine T-lymphocyte subsets obtained after 5 days of monocyte differentiation proved efficient for CD4 or CD8 T-cell purification (?90%). The in vitro analysis using L. chagasi-infected macrophages and purified T cells presented a prospective methodology that could be incorporated in CVL vaccine and treatment studies that aim to analyze the microbicidal potential induced by specific CD4(+) and/or CD8(+) T cells. PMID:24018185

Viana, Kelvinson Fernandes; Aguiar-Soares, Rodrigo Dian Oliveira; Roatt, Bruno Mendes; Resende, Lucilene Aparecida; Silveira-Lemos, Denise da; Corrêa-Oliveira, Rodrigo; Martins-Filho, Olindo Assis; Moura, Sandra Lima; Zanini, Marcos Santos; Araújo, Márcio Sobreira Silva; Reis, Alexandre Barbosa; Giunchetti, Rodolfo Cordeiro



Leishmania mexicana Induces Limited Recruitment and Activation of Monocytes and Monocyte-Derived Dendritic Cells Early during Infection  

PubMed Central

While C57BL/6 mice infected in the ear with L. major mount a vigorous Th1 response and resolve their lesions, the Th1 response in C57BL/6 mice infected with L. mexicana is more limited, resulting in chronic, non-healing lesions. The aim of this study was to determine if the limited immune response following infection with L. mexicana is related to a deficiency in the ability of monocyte-derived dendritic cells (mo-DCs) to prime a sufficient Th1 response. To address this issue we compared the early immune response following L. mexicana infection with that seen in L. major infected mice. Our data show that fewer monocytes are recruited to the lesions of L. mexicana infected mice as compared to mice infected with L. major. Moreover, monocytes that differentiate into mo-DCs in L. mexicana lesions produced less iNOS and migrated less efficiently to the draining lymph node as compared to those from L. major infected mice. Treatment of L. mexicana infected mice with ?-IL-10R antibody resulted in increased recruitment of monocytes to the lesion along with greater production of IFN-? and iNOS. Additionally, injection of DCs into the ear at the time of infection with L. mexicana also led to a more robust Th1 response. Taken together, these data suggest that during L. mexicana infection reduced recruitment, activation and subsequent migration of monocytes and mo-DCs to the draining lymph nodes may result in the insufficient priming of a Th1 response.

Petritus, Patricia M.; Manzoni-de-Almeida, Daniel; Gimblet, Ciara; Gonzalez Lombana, Claudia; Scott, Phillip



Tumors induce a subset of inflammatory monocytes with immunosuppressive activity on CD8+ T cells  

PubMed Central

Active suppression of tumor-specific T lymphocytes can limit the efficacy of immune surveillance and immunotherapy. While tumor-recruited CD11b+ myeloid cells are known mediators of tumor-associated immune dysfunction, the true nature of these suppressive cells and the fine biochemical pathways governing their immunosuppressive activity remain elusive. Here we describe a population of circulating CD11b+IL-4 receptor ?+ (CD11b+IL-4R?+), inflammatory-type monocytes that is elicited by growing tumors and activated by IFN-? released from T lymphocytes. CD11b+IL-4R?+ cells produced IL-13 and IFN-? and integrated the downstream signals of these cytokines to trigger the molecular pathways suppressing antigen-activated CD8+ T lymphocytes. Analogous immunosuppressive circuits were active in CD11b+ cells present within the tumor microenvironment. These suppressor cells challenge the current idea that tumor-conditioned immunosuppressive monocytes/macrophages are alternatively activated. Moreover, our data show how the inflammatory response elicited by tumors had detrimental effects on the adaptive immune system and suggest novel approaches for the treatment of tumor-induced immune dysfunctions.

Gallina, Giovanna; Dolcetti, Luigi; Serafini, Paolo; Santo, Carmela De; Marigo, Ilaria; Colombo, Mario P.; Basso, Giuseppe; Brombacher, Frank; Borrello, Ivan; Zanovello, Paola; Bicciato, Silvio; Bronte, Vincenzo



Nucleic acid distribution pattern as a possible biomarker for metabolic activities of neoplastic cells: a digitally-aided fluorescence microscopy study on normal and neoplastic lymphocytes of acute and chronic canine lymphocytic leukemia  

PubMed Central

Background Metabolic states of neoplastic cells are increasingly being relied upon for diagnostic and prognostic assessment of neoplastic conditions. The nucleic acid distribution pattern of cells in general, in terms of degree of condensation of the nuclear chromatin and overall spread of the nucleic acid within the nuclear and cytoplasmic compartments, can reflect the metabolic state of the cell. This simple but logical concept appears not be put into consideration to date as numerous attempts are being made towards formulating reliable biomarkers for rapid diagnosis, prognosis and subsequent therapeutic interventions for neoplastic conditions. We comparatively evaluated nucleic acid distribution patterns of normal lymphocytes and neoplastic cells of lymphocytic lineage, employing light and fluorescence microscopy procedures, as well as digital imaging analytical methods. Results The results demonstrate distinctiveness in the pattern of nucleic acid distribution for the normal lymphocytes and three lymphocytic neoplastic cell-types of canine lymphocytic leukemia that are categorized as small, intermediate and large neoplastic lymphocytes. Variably-shaped cytoplasmic processes laden with single-stranded nucleic acids (SSNA) were observed for the small and intermediate-sized neoplastic lymphocytes, compared with large neoplastic lymphocytes and the normal lymphocytes; the latter two categories of cells being virtually devoid of similar processes. Prominent cytoplasmic and nuclear clumps of SSNA, indicative of a higher rate of metabolic activity, were also observed within the neoplastic cells compared with fewer and narrower SSNA of the normal cells. Conclusion The comparative relative increases of SSNA in cytoplasmic processes and other cellular areas of small and intermediate-sized neoplastic lymphocytes is reflective of greater metabolic activity in neoplastic cells in general compared with their normal cellular counterparts.

Isitor, Godwin N; Campbell, Mervyn; Nayak, Shivananda B




PubMed Central

Peritoneal exudate lymphocytes from guinea pigs immunized with horse radish peroxidase, dinitrophenyl guinea pig albumin, or ferritin in complete Freund's adjuvant have been shown to be significantly more reactive than other lymphocytes in two in vitro assays of cellular immune function: production of macrophage migration inhibitory factor and antigen-induced lymphocyte proliferation. The enhanced reactivity of peritoneal exudate lymphocytes cannot be accounted for by artifacts introduced by column purification or by the presence of nonlymphoid accessory cells. These observations suggest that the peritoneal exudate lymphocyte pool is a highly enriched source of cellular immune effector cells with specificity directed towards those antigenic determinants to which an animal has been recently exposed.

Rosenstreich, David L.; Blake, J. Thomas; Rosenthal, Alan S.



Comparison of lymphocyte apoptotic index and qualitative DNA damage in yoga practitioners and breast cancer patients: A pilot study  

PubMed Central

Background: Yoga is found to be effective in reducing stress levels and radiation-induced DNA damage, and improving the quality of life, in breast cancer patients. The present study was aimed at comparing the apoptotic index (AI) and DNA damage of advanced yoga practitioners with those of breast cancer patients. Materials and Methods: This cross-sectional pilot study compared three groups (n = 9 each) of age-matched subjects viz. (1) Carcinoma breast patients in stage II or III undergoing radiation therapy after completing three cycles of chemotherapy; (2) Senior yoga practitioners who were practicing asanas, pranayama and meditation daily for more than 10 years; and (3) Normal healthy volunteers. Peripheral blood lymphocytes were isolated, and qualitative DNA damage (QDD) and AI were evaluated by single-cell gel electrophoresis assay. Approximately 500 cells were counted in each case. Number of cells that were normal, undergoing apoptosis, and with DNA damage were categorized and percentages were calculated. Results: Data being normally distributed, one-way analysis of variance (ANOVA) showed significant interaction between groups in AI (P = 0.016) and QDD (P = 0.045). On post-hoc analysis using Scheffe test, AI was significantly lower in non-yoga volunteers as compared with the breast cancer group (P = 0.019) and QDD was significantly lower in yoga practitioners when compared with non-yoga volunteers (P = 0.047). Conclusion: Cellular dysfunction (QDD) requires restorative mechanisms (AI) to restore the system to a balance. The results of this pilot study show trends, which indicate that in ill-health, there is inadequate restorative mechanisms (AI) although dysfunction (QDD) is high. Through regular practice of yoga, cellular dysfunction can be lowered, thus necessitating reduced restorative mechanisms. AI and QDD could also be useful indicators for predicting the three zones of health viz. disease, health, and positive health.

Ram, Amritanshu; Banerjee, Birendranath; Hosakote, Vadiraja S; Rao, Raghavendra M; Nagarathna, Raghuram



Monocyte adhesion molecule expression in interstitial inflammation in patients with renal failure  

Microsoft Academic Search

Background. Patients with renal failure have an increased susceptibility to infections. We therefore studied the recruitment of monocytes and their expres- sion of adhesion molecules CD11b and CD62L at the site of interstitial inflammation in patients with renal failure. Furthermore, we studied if the capacity of monocytes to up-regulate CD11b in interstitial inflam- mation was determined by the interstitial concentration

Elham Dadfar; Joachim Lundahl; Stefan H. Jacobson


Estradiol suppresses vascular monocyte chemotactic protein-1 expression during early atherogenesis  

Microsoft Academic Search

Objective: This study was undertaken to determine whether estrogen down-regulates vascular monocyte chemotactic protein-1 expression during the development of atherosclerosis in vivo and to identify the cellular localization of monocyte chemotactic protein-1 expression under baseline conditions and in response to atherogenic stimuli. Study Design: Female, homozygous low-density lipoprotein-receptor-deficient mice (n = 68) in a C57BL\\/6 background underwent ovariectomy, were implanted

Emre Seli; Umit Ali Kayisli; Belgin Selam; Meltem Seli; Aydin Arici



Impaired monocyte cytokine production in critically ill patients with acute renal failure  

Microsoft Academic Search

Impaired monocyte cytokine production in critically ill patients with acute renal failure.BackgroundPlasma levels of pro- and anti-inflammatory cytokines are predictive of mortality in patients with acute renal failure (ARF). Anti-inflammatory strategies are postulated to be beneficial in treatment. However, there are few studies simultaneously examining monocyte cytokine production and plasma cytokine levels in patients with ARF.MethodsStudy populations consisted of 20





PubMed Central

Immunofluorescent studies using live cells from antibody-forming organs and anti-immunoglobulin antibodies demonstrate two populations of small lymphocytes which are differentiated by the presence or absence of Ig on their surface membranes. Most of the lymphocytes with detectable surface Ig appear to derive from cells of the bone marrow, while most of the Ig-negative lymphocytes derive from the thymus. Thus, adult mice thymectomized, lethally irradiated, and transplanted with bone marrow cells showed a normal number of lymphocytes with surface Ig but were depleted of the Ig-negative lymphocytes. Injection of thymocytes into these mice did not result in an increase in the number of lymphocytes with surface Ig in spleen and lymph nodes. Most of the injected thymocytes could be identified by means of histocompatibility markers. Also, the spleen and lymph nodes of neonatally thymectomized mice contained lymphocytes with surface Ig but were depleted of the Ig-negative lymphocytes. Attempts were made to identify light chains on thymocytes by a sensitive radioimmunoassay. In some experiments no light chains were detected and in others a small amount, i.e. no more than 2.6% of the amount present on spleen lymphocytes, could be detected. Whether these low figures are significant or represent a small amount of serum contamination is not clear as yet.

Unanue, Emil R.; Grey, Howard M.; Rabellino, Enrique; Campbell, Priscilla; Schmidtke, Jon



Apolizumab in Treating Patients With Relapsed or Refractory Chronic Lymphocytic Leukemia or Small Lymphocytic Lymphoma

Noncontiguous Stage II Small Lymphocytic Lymphoma; Recurrent Small Lymphocytic Lymphoma; Refractory Chronic Lymphocytic Leukemia; Stage III Small Lymphocytic Lymphoma; Stage IV Small Lymphocytic Lymphoma



Chronic lymphocytic leukaemias and non-Hodgkin's lymphomas by histological type in farming-animal breeding workers: a population case-control study based on job titles  

Microsoft Academic Search

OBJECTIVES--A population based case-control study was conducted in a highly agricultural area in the north east of Italy to evaluate the association between farming and animal breeding and the risk of developing non-Hodgkin's lymphoma (NHL) and chronic lymphocytic leukaemia (CLL). METHODS--Occupational histories and other data were collected by personal interview on 164 NHLs, 23 CLLs, diagnosed in 1988-90, and on

D Amadori; O Nanni; F Falcini; A Saragoni; V Tison; A Callea; E Scarpi; M Ricci; N Riva; E Buiatti



Radioprotective Effects of Honeybee Venom (Apis mellifera) Against 915MHz Microwave Radiation—Induced DNA Damage in Wistar Rat Lymphocytes: In Vitro Study  

Microsoft Academic Search

The aim of this study is to investigate the radioprotective effect of bee venom against DNA damage induced by 915-MHz microwave radiation (specific absorption rate of 0.6 W\\/kg) in Wistar rats. Whole blood lymphocytes of Wistar rats are treated with 1 ?g\\/mL bee venom 4 hours prior to and immediately before irradiation. Standard and formamidopyrimidine-DNA glycosylase (Fpg)—modified comet assays are

Goran Gajski; Vera Garaj-Vrhovac



What's New in Adult Acute Lymphocytic Leukemia Research?  


... Topic More information about acute lymphocytic leukemia What`s new in acute lymphocytic leukemia research? Researchers at many ... that use newer targeted treatments against ALL. A new lab technique is being studied to help find ...


Swelling-Activated Pathways in Human T-Lymphocytes Studied by Cell Volumetry and Electrorotation  

Microsoft Academic Search

Small organic solutes, including sugar derivatives, amino acids, etc., contribute significantly to the osmoregulation of mammalian cells. The present study explores the mechanisms of swelling-activated membrane permeability for electrolytes and neutral carbohydrates in Jurkat cells. Electrorotation was used to analyze the relationship between the hypotonically induced changes in the electrically accessible surface area of the plasma membrane (probed by the

M. Kiesel; R. Reuss; J. Endter; D. Zimmermann; H. Zimmermann; R. Shirakashi; E. Bamberg; U. Zimmermann; V. L. Sukhorukov



Phase II Study of Flavopiridol in Relapsed Chronic Lymphocytic Leukemia Demonstrating High Response Rates in Genetically High-Risk Disease  

PubMed Central

Purpose Patients with chronic lymphocytic leukemia (CLL) with high-risk genomic features achieve poor outcomes with traditional therapies. A phase I study of a pharmacokinetically derived schedule of flavopiridol suggested promising activity in CLL, irrespective of high-risk features. Given the relevance of these findings to treating genetically high-risk CLL, a prospective confirmatory study was initiated. Patients and Methods Patients with relapsed CLL were treated with single-agent flavopiridol, with subsequent addition of dexamethasone to suppress cytokine release syndrome (CRS). High-risk genomic features were prospectively assessed for response to therapy. Results Sixty-four patients were enrolled. Median age was 60 years, median number of prior therapies was four, and all patients had received prior purine analog therapy. If patients tolerated treatment during week 1, dose escalation occurred during week 2. Dose escalation did not occur in four patients, as a result of severe tumor lysis syndrome; three of these patients required hemodialysis. Thirty-four patients (53%) achieved response, including 30 partial responses (PRs; 47%), three nodular PRs (5%), and one complete response (1.6%). A majority of high-risk patients responded; 12 (57%) of 21 patients with del(17p13.1) and 14 (50%) of 28 patients with del(11q22.3) responded irrespective of lymph node size. Median progression-free survival among responders was 10 to 12 months across all cytogenetic risk groups. Reducing the number of weekly treatments per cycle from four to three and adding prophylactic dexamethasone, which abrogated interleukin-6 release and CRS (P ? .01), resulted in improved tolerability and treatment delivery. Conclusion Flavopiridol achieves significant clinical activity in patients with relapsed CLL, including those with high-risk genomic features and bulky lymphadenopathy. Subsequent clinical trials should use the amended treatment schedule developed herein and prophylactic corticosteroids.

Lin, Thomas S.; Ruppert, Amy S.; Johnson, Amy J.; Fischer, Beth; Heerema, Nyla A.; Andritsos, Leslie A.; Blum, Kristie A.; Flynn, Joseph M.; Jones, Jeffrey A.; Hu, Weihong; Moran, Mollie E.; Mitchell, Sarah M.; Smith, Lisa L.; Wagner, Amy J.; Raymond, Chelsey A.; Schaaf, Larry J.; Phelps, Mitch A.; Villalona-Calero, Miguel A.; Grever, Michael R.; Byrd, John C.



HNK-1 monoclonal antibody (Leu-7) in the identification of abnormal expansions of large granular lymphocytes.  

PubMed Central

Among 12 cases of chronic T lymphoproliferative disorders we observed, six patients showed an expansion of mononuclear cells with azurophilic granules usually referred to as large granular lymphocytes or LGL. Cells obtained from five patients with these abnormal LGL proliferations were studied with several surface markers including their reactivity with the HNK-1 monoclonal antibody reported to be specific for LGL. Cells in four out of five cases were HNK-1 positive. Whereas normal LGL have been reported to be unreactive with several T cell markers, three cases showed the co-existence of HNK-1 and surface markers expressed by T cells. Two cases were characterized by the proliferation of OKT8 cells. Cells from one patient were HNK-1 positive but did not express T or monocytic antigens. These cells were apparently not completely mature since alpha-na