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The pathogenesis of rheumatoid arthritis (RA) is driven by a combined action of genetic and environmental factors, which can upset the balance between the effector and regulatory components of the immune system. An important actor in maintaining such balance is T cells, especially regulatory T lymphocytes (Treg), but the mechanisms behind the functioning of T cell subpopulations and the roles of individual etiological factors in RA have not been fully elucidated. This study aimed to investigate the indices of cell-mediated immunity, especially T- and Treg cells, in RA patients depending on the disease activity and presence of cytomegalovirus (CMV) infection. The expression of membrane and intracellular molecular markers of lymphocytes was estimated by multicolor flow cytometry. The content of antibodies to CMV in blood plasma was measured by enzyme immunoassays. Patients with RA had reliably reduced numbers of cells with the phenotypes CD4+FOXP3+, CD4+CD25+FOXP3+ correlating with the stage of RA activity. RA patients with CMV infection showed a reduction in the number of regulatory T cells (Treg), CD3+ T lymphocytes, CD3+CD8+ cells in peripheral blood. At the same time, RA involved a rise in the level of B cells and CD4+CD25+ Т cells. The level of antibodies to CMV was observed to grow in line with RA activity. Thus, the data obtained suggest that the presence of CMV infection can significantly influence the state of individual lymphocyte subpopulations during RA development.

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BACKGROUND: Rheumatoid arthritis (RA) is an inflammatory rheumatic disease associated with a dysfunction of the T cell-mediated tolerance and leading to the disability of working population. The regulatory CD4' T cells play an important role in the regulation of autoimmunity and can suppress immune responses. With that, there is no consensus on the content of these lymphocytes and their role in the pathogenesis of RA. Objective: The aim of the study was to assess the content ofperipheral blood regulatory Tcells (Treg) according to the expression of membrane markers CD4, CD25, CD127 and intracellular FOXP3 marker, as well as the expression of two functional molecules (CTLA-4 and CCR4) in Treg cells of patients with RA. METHODS: Peripheral blood samples of RA patients (median age 65,5 years [54;68,3) and healthy controls (median age 58 years [44; 66]) were analyzed. Cell count and the expression level of molecules were assessed by flow cytometry. RESULTS: Peripheral blood samples of 36 RA patients and 20 healthy donors were analyzed. The number of the cells with Treg-associated phenotypes CD4⁺CD25hi and CD4⁺CD25hiCD127low/⁻ was higher in RA patients in comparison with healthy donors. Increased levels of RA CD4⁺ T cells expressing FOXP3 were also observed. This may be due to increasing in the number of CD4⁺FOXP3⁺CD25⁻ lymphocytes, whereas the content of RA CD4⁺FOXP3⁺CD25⁺ Treg cells was at the level of the control. The expression of the functional molecule CTLA-4 in Treg cells of patients with RA was not different from the control, while the expression level of the chemokine receptor CCR4 which provides migration of lymphocytes at sites of inflammation and barrier tissues was significantly increased in RA patients. CONCLUSION: Increase in the levels of certain Treg-associated lymphocyte populations were detected in peripheral blood of RA patients. During the natural course of RA, alterations in the level of the chemokine receptor CCR4 might indicate the enhanced lymphocyte migration.

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Data on the frequency of peripheral Treg cells, their functional activity, as well as their relationship to changes in characteristics of cellular immunity in patients with colorectal cancer (CRC, n=42) and healthy donors (n=34) are presented in the article. Assessment of the expression of molecular markers of lymphocytes was performed by multicolor flow cytometry. It was shown that the number of these cells is significantly increased in CRC patients (p<0,05). Probably CD4+CD25+FOXP3+ Treg cells may play a more substantial role in the periphery during the initial period of tumor formation, whereas CD8+FOXP3+ Treg cells are important in the later stages of the disease. In CRC patients, the increase of the functional activity of Treg cells (CD4+CD25+CD127lo/') on the expression of inhibitory molecules CTLA-4 and the marker of cell proliferation Ki-67 was observed. In patients with CRC, the frequency of peripheral Treg cells and the expression level of CTLA-4 were inversely correlated with the number of CD3+CD4+ T-helper cells.

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THE AIM OF STUDY: Assessment of changes in lymphocyte number and some indicators of the immune suppression (frequency of regulatory T cells (Treg), the level of expression of CTLA-4 (cytolytic T lymphocyte-associated antigen 4) and interleukin (IL)-10) in patients with acute pancreatitis (AP). MATERIALS AND METHODS: Peripheral blood samples of 21 patients with AP (including 11 patients with pancreatic necrosis (PN)) were studied for analysis of lymphocyte subsets and 11 blood samples of healthy donors were estimated as control. Evaluation of the expression of molecules of lymphocytes was carried out by flow cytometry. RESULTS: It was found that percentage of CD3+ T cells and their subpopulation of CD4+ T-helper cells in patients with AP were decreased as compared to the control group. Substantial changes were observed in patients with PN: together with decrease in the number of T cells, CD19+ B cells and CD4+ CD25+ activated T-helper cells were also reduced, and the percentage of CD8+ T cells was higher. The immunoregulatory index (CD4+/CD8+) was lower in the group of patients, than in control group. The frequency of Treg cells (CD4+ CD25hi and CD4+ CD25+ CD127lo) was higher in patients with AP than in the control. The expression level of the inhibitory molecule CTLA-4 in the subset of CD4+ CD25hi Treg cells in patients with AP was also increased. Moreover, it was found that the expression level of anti-inflammatory cytokine IL-10 was higher in T-helper cells from patients with AP. CONCLUSION: Changes in cellular immunity (especially patients with PN) werefound in patients wish AP. These changes may indicate the possible development of secondary immunodeficiency. This is probably related to the generation of immune suppression, since the values of the investigated parameters (frequency of Treg cells, the expression of CTLA-4 and IL-10) in patients with AP were higher than in controls.

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Regulatory T cells are crucial for maintaining T cell tolerance to-self-antigens and autoimmune process prevention. They are known to be able to cut down in vitro autologous lymphocyte proliferation in mice. It is suggested that their increased levels are responsible for immunosuppression in human tumor growth. Our investigation was concerned with identifying the extent of mitogen-stimulated T- and B-cell concentration changes as well as phenotypic variation in peripheral blood lymphocyte level in lung, stomach, breast and colorectal cancer. It was found that malignancy involves reduced mitogen (PHA, ConA, PWM)-stimulated lymphocyte proliferation and enhanced peripheral blood CD25+-cell concentration which should be accounted for by regulatory T cell increase. However, both the extent of proliferation changes and CD25+-lymphoid cell concentration considerably depended on tumor localization, which might have an indicator of a varying degree of immunosuppression involved in different neoplasms.

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Patients with gastrointestinal tumors revealed relatively high levels of activated lymphocytes. CD25 expression at later stages was much higher than at earlier ones (I-II). A relationship between CD25, CD71 and CD95 expression was established for metastatic spread of gastrointestinal tumors as well as a correlation between CD71 and CD98 expression for favorable course. A distinct activation phenotype can be identified for peripheral blood lymphocytes in burgeoning tumor which should be taken into account in prognosis.

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Expression of activation antigens increased in patients with gastrointestinal tumors of various clinical courses. The count of CD25+ and CD71+ lymphocytes was high in patients with favorable course of non-progressive tumors. The increase in the expression of activation markers probably reflects adequate functional activity of the immune system in patients with tumors.

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The expression of surface markers of blood lymphocytes was studied in 32 lung cancer patients. Tumor process involved enhanced expression of the activation markers CD25, CD71 and HLA-DR as well as differentiation antigens CD4, CD8, CD22 and CD16. The expression of the markers was higher than in control, irrespective of stage, localization and histological pattern of tumor. It is suggested that increased expression of surface markers should be accounted for by tumor-induced suppression of the immune system in lung cancer.

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Heat-induced hemolysis of erythrocytes was evaluated by the equilibrium method at 54-60 degrees C in patients with lung cancer and donors. Thermal sensitivity of patients differed from that of normal subjects and the seeming energy of erythrocyte lysis activation was lower in patients (230 kJ/mol) than in the controls (345 kJ/mol). Presumably changed structural characteristics of membrane bilayer affected the physicochemical properties of erythrocytes, determining, among other things, thermal resistance of red blood cells.

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The lymphocyte proliferative response to PHA, Con A and PWM was studied in patients with lung tumors (stage I-IV) and other histological types. Disorders in the mitogen-induced lymphocyte activation were identified in more advanced tumors. Certain peculiarities of lymphocyte proliferative activity were found to be determined by histological pattern and localization. A method of evaluation of blasttransformation consequences with due account of lymphocyte functional activity in cancer patients is suggested.

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To compare the accuracy of diagnosis obtained with different diagnostic techniques the authors examined 187 patients with cancer of the lung clinically, roentgenologically, bronchologically, morphologically and immunologically. X-ray made the diagnosis of lung cancer in 85% of the examinees. This diagnosis was confirmed in 87% of the cases. Morphological verification was obtained in 84.4% and 78.2% of patients with central and peripheral cancer, respectively. Additional immunological investigations increased the proportion of accurate diagnoses up to 96.8%. It is concluded that immunological investigations are effective in complex diagnosis of lung cancer.

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The delayed-type hypersensitivity skin test was run with autologous modified lymphocytes, treated with phytohemagglunin, in 218 patients suffering different lung diseases. The three following groups were identified: lung cancer, chronic non-specific disease of the lung and benign lesions in the lung. Cancer patients proved positive to autologous modified lumphocytes in 90.4%. Skin reaction was close to normal in the other groups. Skin reaction dynamics was compared in cases of recurrence and recurrence-free patients in the group of surgical treatment for lung cancer.

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The dependence of immunological reactions of fishes infected with Ichthyophthirius multifiliis infusoria was elucidated. The invasion of the parasite results in the activation of the immune system, increased phagocytosis and a growth of the activity of oxydase enzymes. With the increased number and sizes of parasites a decrease of the above parameters below the control values is observed. Changes of the immunological values are most dynamic at high (20-25 degrees) and they are slower at low (10 degrees) temperatures.

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Prediction of a recurrence of thyrotoxicosis during radioiodine therapy of patients with diffuse toxic goiter (DTG) is a pressing problem of everyday treatment of such patients. Altogether 43 DTG patients were investigated during 12-18 mos. after 131I therapy with the determination of T4, T3, bTSH, delta TSH in the TRH test, peripheral blood rosette forming (RFC), migration index (MI) in the leucocyte migration inhibition reaction, and serum immunoglobulins (IgA, M, G). Changes in the immunological status in parallel with confirmation of autonomous thyroid functioning suggested higher risk of a thyrotoxicosis recurrence making it possible to differentiate the type of observation of a DTG patient following radioiodine therapy. The availability of the above tests allows their use for the prediction of recurrences in everyday clinical practice during therapy of DTG patients with 131I.

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The paper is concerned with a multimodality approach to the assessment of clinical remission in 42 patients after radical therapy of toxic goiter. T4, T3, sigma TSH, delta TSH were determined in the thyrotropin releasing hormone stimulation test (TRH-test); some immunological indices (RFCc, RFCn, RFC1, a stimulation index [SI] LBTR, a migration index [MI] in the LMIT and serum immunoglobulins) were also investigated. The abnormal TRH-test was revealed in 2/3 of the patients (the hypo-TRH-test in 10 and the hyper-TRH-test in 11). A conclusion was made of the appropriateness of the multimodality approach to the assessment of toxic goiter remission after therapy.

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The purpose was to study some indices of immunity in the period of mercazolyl therapy and during DTG remission. T4, T3, TSH and some indices of the immunological status: RFCc, RFCl, RFCn, stimulation index (SI) in the lymphocyte blast transformation reaction, serum immunoglobulins (Ig A, M, G) were studied in 44 patients. In diffuse toxic goiter (DTG) RFCc, RFCl and RFCn were two times and more as less, the Ig G level was raised 1.5-fold. Mercazolyl therapy corrected RFCc, RFCn whereas RFCl remained lowered. The Ig G level correlated with the T4 and T3 levels. In the DTG patients an initial drop of SI (50%) was undetectable. Mercazolyl therapy raised SI by the 4-5th week of treatment (113%). SI returned to normal by the 10-12th week. In DTG remission most of the immunologic indices returned to normal.

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