[Significance of evaluation of CD69 expression by peripheral blood lymphocytes for predicting pregnancy outcome in women with recurrent pregnancy loss]. / Znachimost' otsenki ékspressii CD69 limfotsitami perifericheskoi krovi dlia prognoza iskhodov beremennosti u zhenshchin s privychnym vykidyshem.

The aim of this work was to characterize phenotypically peripheral blood T- and NK lymphocytes expressing an early marker of activation, CD69, and assess the significance of CD69 expression for predicting pregnancy outcome in women with idiopathic reccurent pregnancy loss (IRP) before and after immunocytotherapy (ICT). The study group consisted of 36 patients with IRP who became pregnant after pre-gestational allimmunization, in 30 patients the pregnancy was prolonged to the full term and ended with the birth of a viable baby, in 6 - it was terminated before 12 weeks of gestation. In the control group, 15 fertile women outside pregnancy and 11 women at 12 weeks of physiological pregnancy were examined. Assessment of the CD69 expression in women with prolonged pregnancy revealed the absence of significant differences with the control group in the content and proportion of activated lymphocytes (CD69+). In women with aborted pregnancy after pre-gestational ICT, an increase in the number of almost all analyzed lymphocyte subpopulations responding to the activation stimulus, with a clear tendency to increase the proportion of activated T- but not NK-lymphocytes was found. At 5-6 weeks, the proportion of activated lymphocytes among a subpopulation of cytotoxic T-lymphocytes (CD3+CD8+/CD3+CD8+CD69+) in these women was significantly higher than in women with prolonged pregnancy, which confirms the leading role of effector cytotoxic T-lymphocytes in rejection reactions. Thus, the studies showed the promise of evaluating the expression of the early activation marker CD69 as an additional laboratory criterion for the personable appointment of immunocytotherapy to women with a common reccurent pregnancy loss.

Lymphocyte Activation in the Development of Immune Tolerance in Women with Recurrent Pregnancy Loss.

Association between lymphocyte activation and formation of immune tolerance, as well as pregnancy outcome, in the case of immunocytotherapy (ICT) was studied in women with idiopathic recurrent pregnancy loss (IRPL). The content and phenotypic characteristics of activated T lymphocytes and NK cells were investigated in the peripheral blood of IRPL patients with different pregnancy outcomes (pregnancy prolongation to the full term and habitual miscarriage). The fraction of activated cells in the subpopulation of cytotoxic T lymphocytes (CD3+CD8+/CD3+CD8+CD69+) before ICT was significantly lower in women who lost the pregnancy. After ICT, the fraction of these cells during weeks 5-6 of pregnancy in woman with miscarriage was higher than in women with pregnancy prolonged to the full-term. Excessive content of activated cytotoxic lymphocytes can be a mechanism underlying impaired maternal immunotolerance to fetal alloantigens, which is a leading factor of early pregnancy loss. The obtained data confirm the involvement of activated Th17 cells and FOXP3+ Treg cells in the formation of tolerance to paternal antigens of the fetus. Comparison of the decrease in the fraction of CD4+CD25highRORγt+ lymphocytes at the early gestation period (5-6 weeks) and significant upregulation of the IL-17 production by in vitro stimulated whole blood cells in women with miscarriage with the same parameters in women with prolonged pregnancy suggested an imbalance between pro-inflammatory Th17 cells and Treg cells. No such imbalance in the content effector T lymphocytes was observed in women with the full-term pregnancy. Taken together, our data indicate an important role of gestational activation of lymphocytes in the formation of maternal immune response to fetal alloantigens necessary for the prolongation of pregnancy.

[Autoimmune markers for non-invasive diagnosis of endometriosis in women]. / Autoimmunnye markery dlia neinvazivnoi diagnostiki éndometrioza u zhenshchin.

Endometriosis is a common estrogen-dependent chronic disease in women of reproductive age; it is associated with dysregulation of the immune response, local inflammation, and increased formation of autoantibodies. The aim of the study was to investigate the profile of autoantibodies in women with endometriosis and to evaluate their diagnostic value using new modifications of enzyme immunoassay. In women with endometriosis of stage III-IV (n=39), a wide spectrum of autoantibodies was detected, mainly of class G, including antibodies to endometrial antigens (tropomyosin 3, tropomodulin 3), the enzyme α-enolase, steroid (estradiol, progesterone) and gonadotropic hormones. At the same time, the frequency of detection of IgG antibodies to tropomyosin 3, α-enolase, estradiol and human chorionic gonadotropin and their levels in patients with endometriosis were higher than in healthy women (n=26) (p<0.05). IgG-antibodies to tropomyosin 3, α-enolase and estradiol were characterized by higher diagnostic value for endometriosis. The diagnostic value was significantly increased when these antibodies were combined: the AUC reached 0.875 [0.772-0.978] (p<0.0001), the sensitivity and specificity were 83.3% each. Thus, autoantibodies to tropomyosin 3, α-enolase, and estradiol are promising for inclusion in the panel of biomarkers for non-invasive diagnosis of endometriosis.

Production of active oxygen species by blood phagocytes of pregnant women and their newborns with intrauterine infection.

We studied the relationship between changes in the maternal and newborn granulocyte functions under conditions of infection risk and realization. Women with normal gestation and their healthy newborns, pregnant women with a high risk of infection and their newborns, healthy or with intrauterine infection, were examined. Changes in the active oxygen species-dependent phagocytosis system were found in the blood of risk group patients. An inverse relationship between the parameters venous and umbilical cord blood was detected indicating a relationship between changes in functional activities of maternal and newborn granulocytes. The percentage of CD11b(+)cells in venous and umbilical cord blood strictly correlated with the percent of cells that phagocytosed FITC-labeled E. coli. Deviations in the generation of active oxygen species in phagocytosis seemed to be related to the expression of surface receptors in the risk groups.

Time course of the cytokine profiles during the early period of normal pregnancy and in patients with a history of habitual miscarriage.

The time course of the peripheral blood cytokine profiles was studied in patients with a history of habitual miscarriages and in normal gestation. Low levels of anti-inflammatory cytokines during the early periods of gestation were characteristic of patients with a history of habitual miscarriages; however, an anti-inflammatory shift of the cytokine spectrum developed by the end of the first trimester.

Regulation of reactive oxygen species production by peripheral blood neutrophils from women with postpartum endometritis.

Production of reactive oxygen species in unfractionated peripheral blood increased in parturient women without postpartum infectious complications and patients with postpartum endometritis. The control group included nonpregnant women with normal reproductive function. Intergroup differences were revealed in the degree of respiratory burst activation with opsonized zymosan and response of isolated granulocytes to chemotactic peptide N-formyl-Met-Leu-Phen (1 muM). Production of reactive oxygen species tended to normal after therapy. We studied the effects of a specific mitogen-activated protein kinase p38MAPK inhibitor and inhibitors of tyrosine protein phosphatases and phosphatidylinositol-3-kinase. The role of p38MAPK in reactive oxygen species generation by cells changes significantly in parturient women.

[Immune system in women with genital inflammatory diseases caused by Chlamydia and virus infection]. / Osobennosti sostoianiia immunnoi sistemy u zhenshchin s vospalitelnymi zabolevaniiami genitalii Khlamidiinoi i virusnoi étiologii.

Immunologic studies were carried out in 29 women with clinical manifestations of chronic inflammations of the internal genitals. Dot-hybridization with biotin-labeled DNA probes revealed chlamydial DNA in the cervical cells of 14 women, Herpes simplex type 2 and/or cytomegaloviral DNA in 15 women. Cell-mediated immunity was assessed using monoclonal antibodies CD3, CD4, CD8, and CD19 and by Facscan flow cytofluorometer (Becton Dickinson). Immunoglobulins were measured by nephelometry with Abbott kits and TDx device. No appreciable changes in the mean values of the parameters characterizing T-cellular immunity were revealed in women with genital inflammations and chlamydial infection (p < 0.05). The counts of CD19+ lymphocytes varied to a certain degree, and IgM levels were increased in patients with chlamydial infection, this pointing to activation of B-lymphocytes with Chlamydia. A viral infection of the genitals was associated with a reduction of T-cellular immunity parameters and an imbalance in the ratio of immunoregulatory cells at the expense of reduction of the share of lymphocytes expressing CD4 antigen.

Studies of the immunologically specific retention of antigen-activated lymphoid cells in antigen-containing lymph nodes.

Mouse spleen lymphocytes proliferating under the influence of SRBC, or educated thymocytes were labelled in vivo or in vitro with 3H-thymidine and transferred intravenously to syngeneic recipients, which had received a subcutaneous injection of the same, or a non-cross-reacting, antigen (SRBC, rat or chicken RBC, DNP-proteins) into the right front footpads. The left footpads had been injected with a control (non-cross-reacting) antigen. The right (ipsilateral) and left (contralateral) regional lymph nodes were compared on the basis of their radioactivity and radioautography. SRBC-containing lymph nodes retained specifically both viable primed spleen cells and educated thymocytes. Treatment of donor spleen cells with anti-T serum and complement prevented specific retention. Administration of anti-SRBC serum to the recipients had a similar effect. Retention occurred during 6 days after administration of the antigen to the recipients and was mediated by radioresistant host lymph node cells. It declined profoundly when host macrophages were blocked with charcoal particles. The data obtained suggest a major role of the direct contact between T cell receptors and macrophage-bound antigen in the specific lymphocyte retention.

Bone marrow suppressor B cells in vitro.

Marrow cells from intact CBA and (CBA x C57B1)F1 mice added to a culture of syngeneic splenocytes at its initiation suppressed the generation of anti-SRBC antibody-forming cells. Removal of cells bearing surface immunoglobulins and/or MBLA led to a substantial reduction in the marrow suppressive activity. Marrow cells from mice pretreated with five doses of hydroxyurea did not suppress the immune response of syngeneic splenocytes to SRBC. While the total number of cells in the marrow of such animals declined 5-fold, the relative content of Ig-positive cells was somewhat increased. The proportion of blast cells dropped from 10.3 to 2.4%. No blast cells bearing surface immunoglobulins were observed. The proportion of cells incorporating [3H]-thymidine fell 40-fold. We conclude that suppression of the immune response to SRBC in vitro is mediated by immature precursors of the B lymphocyte series which are present in the bone marrow of intact mice.