Jacek Tabarkiewicz

The predominance of Th17 lymphocytes and decreased number and function of Treg cells in preeclampsia.

The aim of this study was to estimate the prevalence of CD3(+)CD4(+) T lymphocytes producing IL-17, IL-2, IFN-γ, and IL-4, plus CD4(+)CD25(+)FoxP3(+) T regulatory (Treg) cells, in peripheral blood of patients with preeclampsia and healthy women in the third trimester of normal pregnancy. Another purpose was to assess the immunosuppressive activity of Treg cells from patients with preeclampsia compared with controls. Thirty-four preeclampsia patients and 27 healthy pregnant women were included. The percentages of CD4(+)CD25(+)FoxP3(+) Treg cells and CD3(+)CD4(+) T lymphocytes with intracellular expressions of cytokines were estimated using monoclonal antibodies and flow cytometry. In vitro functional assays were performed using a Treg Cell Isolation Kit and (3)H-thymidine incorporation assays. The percentage of CD3(+)CD4(+) T lymphocytes producing IL-17A was significantly higher in preeclampsia than in healthy, normotensive pregnant women in the third trimester (p<0.001). The population of CD4(+)CD25(+)FoxP3(+) Treg cells was significantly lower in the study group compared with controls (p<0.05). There was no change in the stimulation index of CD3(+)CD4(+)CD25(-) T lymphocytes from preeclampsia patients without Treg cells and after addition of autologous Treg cells. In normal pregnancy, the stimulation index of CD3(+)CD4(+)CD25(-) T lymphocytes was significantly higher without Treg cells compared with the response after addition of autologous Treg cells (p<0.05). The results suggest up-regulation of the Th17 immune response in preeclampsia. The decreased number and function of Treg cells may be responsible for activating the inflammatory response characteristic of this disorder. In preeclampsia, the predominance of Th17 immunity could act through modulating the Th1/Th2 immune balance.

Activated T lymphocytes in pre-eclampsia.

The aim of our study was to investigate the activation markets of T CD3+, T helper CD4+ and T cytotoxic CD8+ cells, as well as, the populations of T naïve CD4+ CD45RA+, T memory CD4+ CD45RO+ and T regulatory lymphocytes in PE and healthy pregnant women.

Blood myeloid and lymphoid dendritic cells are stable during the menstrual cycle but deficient during mid-gestation

The aim of this study was to estimate the populations of peripheral blood myeloid and lymphoid dendritic cells (CD1c(+), BDCA-2(+), BDCA-4(+)) and the CD1c(+):BDCA-2(+) ratio in phases of the ovarian cycle and in normal pregnant patients. 18 non-pregnant women and 17 normal pregnant women were included. Dendritic cells were isolated from peripheral blood, stained with monoclonal antibodies (mAbs) against blood dendritic cell antigens (anti-BDCA-1, BDCA-2, BDCA-4) and estimated using flow cytometry. CD1c(+), BDCA-2(+) and BDCA-4(+) dendritic cells were present in the follicular and luteal phases of the ovarian cycle and in all trimesters of normal pregnancy. The percentages of CD1c(+) dendritic cells did not differ between the follicular and luteal phases of the ovarian cycle. The percentage of BDCA-2(+) dendritic cells was lower in the luteal phase of the ovarian cycle compared with the follicular phase, but the differences were not statistically significant. The CD1c(+):BDCA-2(+) cell ratio was significantly lower in the luteal phase compared with the follicular phase of the ovarian cycle. The numbers of dendritic cells were significantly lower in the second trimester when compared with the first and third trimesters of normal pregnancy. Furthermore, in the second trimester, the CD1c(+):BDCA-2(+) ratio was higher than in the other trimesters of normal pregnancy. All populations of dendritic cells and the CD1c(+):BDCA-2(+) ratio did not differ in the first and third trimesters of physiological pregnancy. Our results suggest that myeloid and lymphoid dendritic cells are not affected by steroid hormones during the menstrual cycle. The deficiency of peripheral blood dendritic cells observed during the second trimester of normal pregnancy can be associated with their migration to the uterus during the second physiological invasion by cytotrophoblast.

Immunomodulatory Effects of IFN-β and Lovastatin on Immunophenotype of Monocyte-Derived Dendritic Cells in Multiple Sclerosis

Multiple sclerosis (MS) is an autoimmune disease of the central nervous system and current MS treatment is only partially effective. Recent data suggest that statins may be potent immunomodulatory agents. In order to evaluate their role in MS, we analyzed the in vitro effects of interferon (IFN)-β and lovastatin on the differentiation and maturation of monocyte-derived dendritic cells (DCs) of MS patients. Twenty-seven patients with relapsing–remitting MS were recruited for the study. DC differentiation and maturation were evaluated based on surface phenotypic changes and the expressions of CD14, CD83, CD1a, CD80, CD86, CD206, and C209 were analyzed by flow cytometry. The results showed that IFN-β and lovastatin affect DC phenotype. Both agents decrease the expression of CD1a, which indicates a weakened presentation of glycolipid antigens. IFN-β causes up-regulated and lovastatin down-regulated expression of CD86, which results in a biased Th-cell responses in MS. Furthermore, high doses of lovastatin cause a decrease in CD209 expression on the surface of DCs and can limit their migration to various tissues. One of the mechanisms of the beneficial action of IFN-β and statins may be associated with their influence on DCs.

The expression of B7-H1 and B7-H4 co-stimulatory molecules on myeloid and plasmacytoid dendritic cells in pre-eclampsia and normal pregnancy

The aim of our study was to estimate the expression of B7-H1 and B7-H4 molecules on myeloid and plasmacytoid dendritic cells (DCs) in the peripheral blood of patients with pre-eclampsia, normal pregnant women and healthy non-pregnant women. Thirty-three patients with pre-eclampsia, 26 normal pregnant women, and 12 healthy non-pregnant women were included in the study. Dendritic cells were isolated from peripheral blood, stained with monoclonal antibodies against blood dendritic cell antigens and B7-H1 and B7-H4 molecules and estimated using flow cytometry. The expression of B7-H1 and B7-H4 molecules was significantly higher on CD1c(+) myeloid and CD303(+) plasmacytoid DCs in the first trimester of pregnancy than in the luteal phase of the ovarian cycle (CD1c(+)B7-H1(+): 19.19±10.55% vs. 11.99±6.79%; p<0.05; CD1c(+)B7-H4(+): 12.01±9.15% vs. 3.98±1.97%, p<0.001; CD303(+)B7-H1(+): 4.15±2.38% vs. 1.70±0.87%, p<0.05; CD303(+)B7-H4(+): 5.44±2.93% vs. 2.33±1.54%, p<0.01). Moreover, the expression of the B7-H1 molecule on CD1c(+) DCs in the second trimester of normal pregnancy was significantly higher than in the first trimester, but in the third trimester they decreased compared with the second trimester (II vs. I trimester: 32.23±11.30% vs. 19.19±10.55%, p<0.01; III vs. II trimester: 32.23±11.30% vs. 22.39±8.19%, p<0.01). The expression of B7-H1 molecule on CD1c(+) myeloid and CD303(+) plasmacytoid DCs was significantly lower in pre-eclampsia than in healthy third-trimester pregnant women (CD1c(+)B7-H1(+): 13.78±6.26% vs. 22.39±8.19%, p<0.05; CD303(+)B7-H1(+): 3.66±2.46% vs. 8.65±3.15%, p<0.01). Higher expressions of B7-H1 and B7-H4 molecules on CD1c(+) myeloid and CD303(+) plasmacytoid DCs in the first trimester of pregnancy suggest the role they play in the immunomodulation during early pregnancy.

The expressions of co-stimulatory molecules are altered on putative antigen-presenting cells in cord blood.

The aim of our study was to estimate the expressions of both B7‐H1 and B7‐H4 as well as CD200 and CD200R co‐stimulatory molecules on immature myeloid and lymphoid dendritic cells, B CD19+ lymphocytes and monocytes in umbilical cord blood of healthy neonates and in peripheral blood of healthy adults.

Dendritic cell subsets in neoplastic tissue and peripheral blood of laryngeal cancer patients: relation with grade and stage of the disease

Despite improvements in immunotherapy, little is known about dendritic cell (DC) subpopulations naturally occurring in the laryngeal cancer tissue (LCT) and peripheral blood (PB) of untreated laryngeal cancer (LC) patients. The purpose of the present study was to evaluate mature, immature myeloid and plasmacytoid DCs in the LCT and PB of patients with various stages and grades of squamous cell carcinoma of the larynx (n=66) and PB of healthy donors (n=20), and to explore the correlation of the percentages of the DCs to clinical parameters. The percentage of DCs in LCT and PB was determined using monoclonal antibodies and flow cytometry. It has been revealed that DCs accumulate in LCT in comparison with their content in PB. The myeloid to lymphoid/plasmacytoid DC ratio was higher in LCT compared to PB. It was found that in cases of poorly-differentiated LC, there were higher percentages of lymphoid/plasmacytoid DCs in LCT in comparison to their content in the PB. Moreover, in the blood of patients with T4 cancers we found significantly lower percentages of myeloid DCs in comparison to individuals with T1 neoplasms. The percentage of myeloid DCs infiltrating cancer tissue positively correlated with the T stage. In patients with no metastases in the lymph nodes, PB contained less mature DCs but higher amount of myeloid DCs compared to LCT. Alteration of the DC proportion in LC patients may result in the development of immunotolerance.

OS007. The apoptosis markers are altered in CD4(+)CD25(+)FOXP3(+) T regulatorylymphocytes in pre-eclampsia.

INTRODUCTIONnPre-eclampsia (PE) is a common obstetric syndrome affecting about 5-10% of pregnant women. The etiology and pathogenesis of this syndrome are not fully understood. There are many studies describing alterations in the innate and adaptive immune system which may have an influence on the onset of this disorder. It was suggested that the activation of cell-mediated immunity may play the key role in the etiology of pre-eclampsia. It was proposed that inappropriate activation of the immune system can lead to pre-eclampsia.nnnOBJECTIVESnThe aim of our study was to estimate the surface expressions of CD95(APO-1/Fas) antigen and the intracellular expressions of anti-apoptotic proteinBcl-2 and pro-apoptotic proteinBax in CD4(+)CD25(+)FoxP3(+) T regulatory lymphocytes (Tregs) as well as the percentage of CD8(+)CD28(+) T cytotoxic cells in peripheral blood of patients with pre-eclampsia in comparison with healthy pregnant women in the third trimester of physiological pregnancy.nnnMETHODSnTwenty-four women with pre-eclampsia and twenty normal third trimester pregnant women were included in the study. The lymphocytes were isolated from peripheral blood samples and labelled with monoclonal antibodies. The expressions of surface antigens and intracellular proteins were estimated using flow cytometry.nnnRESULTSnThe population of CD4(+)CD25(+)FoxP3(+) Treg cells was significantly lower in peripheral blood of patients with pre-eclampsia when compared to normal third trimester pregnant women. The percentages of CD4(+)CD25(+)FoxP3(+) Treg cells that express Bcl-2 protein were significantly lower in peripheral blood of patients with pre-eclampsia when compared to healthy pregnant women, whereas the percentages of CD4(+)CD25(+)FoxP3(+) Treg cells with the expressions of Bax protein did not differ in both groups. Moreover, the mean fluorescence intensity (MFI) of Bcl-2 protein in CD4(+)CD25(+)FoxP3(+) Treg cells was significantly lower and MFI of Bax protein significantly higher in pre-eclampsia when compared to the control group. The percentage of CD8(+)CD28(+) T cells did not differ in both studied groups but MFI of CD28 antigen on T CD8(+) cells was significantly higher in pre-eclampsia when compared to the control group.nnnCONCLUSIONnThe obtained results suggest that the deficit of CD4(+)CD25(+)FoxP3(+) Treg lymphocytes which is observed in pre-eclampsia maybe associated with alterations in apoptosis markers.

O3. The predominante of Th17 lymphocytes and decreased number and function of Treg cells are present in preeclampsia.

dictors of severity of preeclampsia, in Colombian population M.C. Páez , N.C. Serrano , L.A. Díaz , M.A. Beltrán , R. Ortíz , A. Monterrosa , W. Saldarriaga , C. Mesa , J.E. Sanin , G. Monsalve , E. Guío , P.K. Bautista , C. Colmenares , C. Vargas , M.E. Hincapie , E. Staines , A. Hingorani , J.P. Casas h,i ( Biomedical Research Center, Faculty of Health, UNAB, Colombia, b Universidad de Cartagena, Colombia, c Universidad del Valle, Colombia, d Universidad CES, Medellín, Colombia, e Universidad Pontificie Bolivariana, Medellín, Colombia, f Clínica del Prado, Medellˇ́n, Colombia, g Universidad Industrial de Santander – UIS, Bucaramanga, Colombia, h Faculty of Epidemiology and Public Health, London School of Hygiene & Tropical Medicine, UK, i Department of Epidemiology and Public Health, University College London, UK)