The effect of β-galactoside-binding protein from lectins family galectin-1 on the subpopulation structure and immunosuppressive function of CD4 +-regulatory lymphocytes was studied in vitro. The action of galectin-1 on the differentiation of CD4+CD25+/CD4 +CD25 +Foxp3 + cells depends on the ratio of lymphocytes maturity and conditions of cell cultivation. The incubation of pre-differentiated cells with the recombinant galectin-1 was accompanied by the increase in the number of lymphocytes with CD4 +CD25 +Foxp3 + phenotype, the increase in the content and expression level of transcription factor Foxp3 mRNA and the increase in cytolytic protein perforin production. The findings suggest that the immunoregulatory effects of galectin-1 are based on the maintaining of the regulatory phenotype of differentiated CD4 +-lymphocytes. Experimental model that we have developed, can be used to regulate the process of lymphocyte differentiation toward regulatory T-cell and to receive the populations of cells with immunosuppressive properties.

Aim of study: to examine the features of expression of CD-markers of blood mononuclear leukocytes and their functional activity in the metabolic syndrome. Conducted a cross-sectional (transverse) study of 76 patients with essential hypertension (EH) II stage (BP <180/110 mm Hg.) [10] in conjunction with the metabolic syndrome and 20 people, formed the control group. Along with a complete clinical, laboratory and instrumental examination taken in a specialized cardiological clinic was conducted determination of surface markers of lymphocytes CD4+, CD8+ and monocytes CD36+ and assessment of the level of spontaneous production of reactive oxygen blood mononuclear leukocytes. Found that in patients with the metabolic syndrome compared with the control group the proportion of CD4+ lymphocytes and the level of spontaneous ROS production by mononuclear leukocytes significantly higher. The positive correlated interconnection between these indicators and the number of CD36+ monocytes with the majority of clinical and metabolic markers of MS confirmes their participation in mechanism of immune inflammation and oxidative stress in this pathological process.

The aim of the study: identification of molecular mechanisms of hydrogen sulfide influence on cell cycle progression. Material and methods: Jurkat cells were the material of the study. Methods included flow cytometry, western blotting, RTPCR. Results and discussion: The delay of G1 cell cycle phase connected with the decrease in content of phosphorylated form of retinoblastoma protein in 50 μM NaHS-treated cells. Phosphorylation of retinoblastoma protein takes place when cyclins activate cycline-dependent kinases. It has been shown that hydrogen sulfide in low concentration leads to decrease of cycline D1 level via decrease of respective gene expression.

Aim. To experimentally select the optimum concentration of hydrogen peroxide capable of efficiently induce oxidative stress and launch the programmed death of the maximum number of lymphocytes, but not induce the necrosis. Methods. Jurkat tumor cell line (human T-lymphoblastic leukemia) lymphocytes isolated from the blood of healthy donors (15 males, 18 females) aged 18 to 25 years were the objects of the study. To confirm the object of study, blood cells typing for CD5 using flow cytometry was performed. To model the oxidative stress in vitro, blood lymphocytes were incubated in the presence of hydrogen peroxide at a final concentration of 0.3, 0.5, 1.0 and 2.0 mM. Reduced and oxidized glutathione levels estimation, the ratio between the fractions, and the level of reactive oxygen forms in lymphocytes for a relative assessment of the oxidative stress degree in cancer cells, were used. Results. An optimal final concentration of hydrogen peroxide — 0.5 mM — was established, causing an increase of active oxygen forms concentration in cells, comparable to levels in tumor cells, the formation of a maximum number of annexin positive cells and minimum number propidium-positive cells and the comparable ratio of the reduced and oxidized glutatione levels. Conclusion. The optimum concentration of hydrogen peroxide (0.5 mM) was selected for the oxidative stress formation in the peripheral blood lymphocytes to study the apoptosis dysregulation in oxidative stress in Jurkat line tumor cells (human T-lymphoblastic leukemia).

The influence of CO-releasing compound CORM-2 on the key molecules regulatirs of the cellular cycle G1 phase (cyclin D1, retinoblastoma protein, protein-inhibitor р21) of Jurkat line has been analyzed in the given paper. It is shown that CO donor causes the delay of cells in G1 of the cellular cycle against the background of increasing the content of cyclin D1 and phosphorylated form Rb. Antiproliferative effect of carbon monoxide was not mediated by increasing the content of protein р21.

In this paper we investigated p38-dependent mechanisms of gaseous transmitters nitric oxide and hydrogen sulfide influence on XIAP and AVEN genes expression in Jurkat cells. Using the method of real-time PCR, XIAP and AVEN genes expression in cancer Jurkat cells was shown. Donors of gases of nitric oxide and hydrogen sulfide were negative regulators of the XIAP and AVEN gene expression in the cells of T-lymphoblast leukemia. Changes in the expression of above mentioned genes under NO and H2S influence occurred with the involvement of p38 MAP kinase

In this paper, participation of gases, nitric oxide, carbon monoxide and hydrogen sulfide, in cell apoptosis regulation has been analyzed according to the literature data and our own findings. Different mechanisms of nitric oxide influence on apoptotic reaction including modulation of transcription factors activity and increase in mitochondrion membrane permeabilisation are described. Brief description of the generation and signal transduction pathways of carbon monoxide is presented. Pro- and antiapoptotic mechanisms of hydrogen sulfide influence on cell fate are analyzed.

Main molecular targets of nitric oxide, hydrogen sulfide and carbon monoxide proapoptotic action in Jurkat cells were determined in this study. Decrease of mitochondrial transmembrane potential was shown during all three gases action. Reason of this event is the Bcl-2 family members disbalance. Proapoptotic proteins release after mitochondrion membranes permeabilisation could be abolished by protein xIAP inhibition of caspase -9 and -3 activity during NO and CO application.

The in vitro phosphorylated and non-phosphorylated Hsp27 forms concentrations and Bcl-2 proteins affected by Hsp27 inhibition were studied in Jurkat-line tumor cells and healthy donor mononuclear lymphocytes by Western blotting technique. The Hsp27 inhibition causes the increase of intracellular Bax protein concentration and the decrease of Bcl-2 level leading to an increase of apoptotic changes in Jurkat line cells.

Cytokines and their receptors belong to a significant role in the initiation and the subsequent course and outcome of autoimmune thyroid disease. Interleukin-2 (IL-2), interleukin-4 (IL-4) and tumor necrosis factor-alpha (TNF-α)-cytokines, which have a multifaceted impact on the various stages of the immune response: the development of inflammatory response, cell proliferation, antibody and acute phase proteins synthesis. Pre-existing pattern of development of autoimmune thyroiditis (Hashimoto's thyroiditis) and Graves' disease (GD) as a state with two opposite positions of the predominant profile of Th1/Th2-lymphocyte activation. The study evaluated the cytokine production by Th1and Th2-lymphocytes in patients with GD, assessment of lymphocyte receptor system and identified lymphocytes subpopulation in patients with BG, and the impact on the functional state of thyroid gland. It was shown that the immunoregulatory cytokines as Th1(IL2)and Th2(IL-4)-helper lymphocytes are involved in the immune mechanism of BG. The level of IL-2, IL4, and TNF-α, and the number complementary lymphocyte receptors were not significantly changed in euthyroid or hyperthyroid GD patient. Nevertheless, there are strong correla! tions between production of immunoregulatory cytokines (IL-2, IL-4) with the functional state of the thyroid gland and increase of its volume in GD patient, what confirms the “functional synergies” of these cytokines in autoimmune inflammation in the GD.