Abstract. The purpose of this article was to make a broad audience of experts familiar to new developments in the method of flow cytometry that have found applications in medical and biologic studies, in order of their further implementation into everyday practice. These applications include the following approcahes: detection of antigen-specific T-cells by using of tetramer technology, flow cytometric determination of cytokines in biological liquids, determination of in vitro sensitivity of basophilic granulocytes to allergen effects, detection of Th1 and Т-reg cells by their cell surface markers. When using multiparametric analysis, a multi-step gating and other new technologies, the flow cytometry technique allows of location and tracing the majority of processes occurring in development of immune response. When studying these dynamic events, we get an opportunity to react adequately to apropriate changes, and to develop new approaches to correct altered cellular activities, that they should determinate.

Abstract. Immunomodulating drugs play an important role in therapy and prevention of numeous diseases associated with altered immune functions. Immunomodulators of bacterial origin are the most active ones, serving as a basis for design of therapeutic vaccines that are capable of stimulating antigen-specific response, along with nonspecific actions. The Immunovac VP4 poly-component vaccine is among such preparations, being a potent activator of innate immunity and showing protective activities against a number of facultative pathogens. In present review, the data are summarized that concern therapeutic effects of Immunovac VP4 in various disorders (lung abscess, chronic bronchitis, bronchial asthma, atopic dermatitis, pyodermia, herpes, acute respiratory infections). In all cases, high clinical effect was registered, i.e., decrease in number and severity of relapses, decreased dosage/number of medical drugs applied, prolongation of remission states, and transition to less severe clinical forms. The therapeutic effect is accompanied by sufficient positive dynamics of immunological parameters, e.g., phagocytic activity of macrophages, increase in lymphocytes bearing CD4, CD8, CD16, CD72, CD21 markers, enhanced IFNγ and IFNα production, correction of Ig synthesis, increased antibody titers and affinity. Analysis of data from strictly controlled studies performed in limited clinical samples, has shown a number of general regularities for common effects of microbial antigens in various disorders including allergic diseases.

Abstract. We examined metabolic status of blood lymphocytes in forty-four patients with chronic myeloid leukemia (CML), and in fifty-seven patients with chronic lymphoid leukemia (CLL). The most pronounced changes of enzyme activities are found in blood lymphocytes of CML and CLL patients during terminal stage of the disease. All the patients with chronic leukemia at both stages showed a decreased intensity of metabolic processes in blood lymphocytes. In addition, CML patients at both clinical stages exhibited a decrease in antioxidant cell protection. The more pronounced biochemical disturbances in CML patients at progression stage concerned carbohydrate metabolism, whereas the changes at terminal stage of disease affected both carbohydrate and fat metabolism. The most marked biochemical alterations in CLL at progression stage are registered for carbohydrate and protein metabolism, and carbohydrate, fat and protein metabolism at terminal stage of disease. The revealed changes showed indicate to more severe clinical course in the patients with CML. These changes and differences depict some aspects of immune pathogenesis in development and progression of CML and CLL.

Abstract. The aim of present study was to analyze the most commonly detected blood plasma immune proteins, that reflect molecular pathways in development of infectious endocarditis of native valves, together with analysis of marker proteins of disease on the base of modern technological platform for clinical proteomics. Ninety-nine patients with infectious endocarditis of native heart valves were included into the study. All the patients were examined according to hospital assessment protocol, including clinical, laboratory and instrumental methods of evaluation. Separation and identification of proteins of microorganisms in blood and materials taken at surgery, detection of proteins related to functioning of immune system, were carried out using new methods of clinical proteomics. We have revealed same proteins, that are specific of pathogens causing infectious endocarditis, that thus representing immunogenic determinants and defining a systemic immune response in the patients.

Abstract. Features of local immunity were studied in the patients with Salmonella infection at reconvalescence, both without carriership, and in reconvalescent carriers of microbial flora (RCM). In RCMs, as compared to non-carriers, some alterations in gut microbiocenosis were revealed, along with local deficiency of antimicrobial defense factors, i.e., lactoferrin, lysozyme, secretory IgA, free secretory component, antibodies to Salmonella O-antigen, IgA, as opposed to high levels of IgM, IgG (due to IgG1 and IgG4), cytokines (IL-6, IL-8, IFNγ, IL-4). It was shown that sIgA and IgA deficiency are connected with their mangling local production, whereas the revealed increase in IgМ and IgG concentrations proved to be a consequence of increased local synthesis. Application of polyoxidonium (PO) in RCMs lead to improvement in the gut microenvironment, thus reflected by decreased contamination with facultative flora, disappearance of Proteus, Candida spp. and recovery of indigenous anaerobic flora in 92 per cent of cases. Positive dynamics of local immunity associated with PO treatment was justified by restoration of total IgA and sIgA concentrations, due to enhanced local synthesis, increase in free secretory component, higher titers of antibodies to Salmonella O-antigen, and changes in cytokine contents, with restoration of Th1/Th2 cytokine balance. Under in vitro conditions, a PO-associated suppression of anti-complement Salmonella activity was registered, thus rendering positive effects of the drug towards symbionts in the «host-parasite» system, and shortening the terms of microbial excretion in Salmonella carriers.

Abstract. The aim of this study was to elucidate a role of brain hemispheres in formation of rheumatoid arthritis (RA). The parameters of higher nervous activity, autonomous and immune systems in these patients that were characterized by domination of the left or right hemispheres of a brain, were defined in present work. The results of this work allow of justifying the following items: a) relative increase in functional activity of right brain hemisphere in woman may represent a factor that either contributes to triggering of rheumatoid arthritis, or predisposes for its development; b) formation of rheumatoid arthritis in females is accompanied by complex changes in psychophysiological and immune parameters, that exhibit significant features depending on functional asymmetry of the hemispheres; c) a pronounced dependence is revealed between clinical course of disease and functional asymmetry of hemispheres.

Abstract. To study influence of microenvironment upon functional activity of B cells, we used experimental models of adoptive cell transfer from CBA to congenic CBA/N mice lacking CD5+ B-1 cells, and cocultivation of CBA/N splenocytes with spleen, or peritoneal CBA cells. B cell activity was determined as numbers of IgM-producing cells, and as amounts of cells producing antibodies to a T-independent antigen type 2 (polyvinylpirrolidone). In vivo distribution of transferred cells was determined as the numbers of cells stained with a vital dye (CDFA-SE) in spleen and peritoneum of recipients. Intravenous injection of CBA splenocytes resulted into a significant (3- to 4-fold) increase in numbers of IgM-producing cells in the spleens of xid–recipients, where their levels reached those of CBA mice. Intravenous injection of CBA/N splenocytes into xid-mice did not induce any increase of IgM-producers in their spleen. That means that increased number of IgM-producers in recipient spleen is due to donor cells, presumably, CD5+ B-1 lymphocytes. Meanwhile, restoration of immune response to polyvinylpirrolidone in xid-mice following transfer of CBA splenocytes was not successful. IgM-producing cells were undetectable in peritoneum of intact mice (both CBA and CBA/N). Intraperitoneal transfer of CBA splenocytes also did not induce their accumulation. It could mean that peritoneal microenvironment inhibits B cell activity. Meanwhile, intravenous injection of «silent» peritoneal cells into CBA/N mice brought about great increase of IgM-producers in recipient spleen, i.e., the «job» of B cells was permitted in other microenvironment. The results yielded in vivo are in agreement with data of in vitro experiments. Addition of CBA splenocytes or peritoneal cells (10-50%) to CBA/N splenocytes induced sharp increase of IgM-producing cells in the cultures. The data obtained provide evidence for a decisive role of microenvironment in functional activity of murine B lymphocytes.

Abstract. 72 patients and 30 healthy children of the age of 7-17 years were examined, aiming for substantiation of thymaline, vilon and ventramine application in treatment of chronic gastroduodenitis in children. Disbalance of cellular and humoral immunity and reduction of intensity of lymphocyte-platelet adhesions were established. An increase of IL-1β, IL-2, TNFα and IFNα concentration was registered in absence of IL-4 changes. Application of thymaline, vilon and ventramine essentially improves clinical course of disease, results in normalization of immune profile, reduced levels of proimflammatory cytokines, restoration of lymphocyte-platelet adhesions.

Abstract. The purpose of present work was to study the peculiarities of cytokine production in patients with malignant tumors, as exemplified by lymphoma and malignant melanoma. Sixty-five patients with malignancies (16 to 74 years old) have been surveyed. Contents of IL-1β, IL-4, TNFα and lactoferrin were studied in blood serum using enzyme immunoassay technique. The results of study have shown that the patients with melanoma have a significant reduction in IL-1-β and TNFα in blood serum, thus being suggestive for development of deficient immune response. In malignant lymphoma, the patients did also exhibit subnormal values of TNFα and IL-1β (below a threshold level of 5 pg/ml), thus supposing some deficiency in proinflammatory cytokine secretion. An interrelation was shown between the contents of lactoferrin and proinflammatory cytokines. After radical treatment of melanoma, like as in lymphoma patients free of В-symptoms, the lactoferrin contents were within normal limits, whereas in the patients with generalization of malignancy, and in lymphoma patients with В-symptoms, a trend to its increase has been established. Determination of the mentioned parameters in patients with lymphoma and malignant melanoma would provide useful diagnostic and predictive criteria in these clinical conditions.

Abstract. This study has shown that intravenous normal human immunoglobulin (IVIG), when included into combined therapy of acute epiglottitis, exerts positive effect upon clinical course of the disease and contributes to normalisation of immune status. Due to IVIG therapy, an increase in IgA and sIgA concentrations is observed in laryngopharynx secretions. Amounts and affinity of serum antibodies to common bacterial antigenic determinant are increased in blood, as well as opsonizing properties of blood serum. Moreover, a higher phagocytic and bactericidal activity of neutrophils is observed with such treatment. Inflammatory as well as degenerative and destructive processes in epiglottis seem to be preventable under the influence of IVIG therapy.

Abstract. Collagen type II (Col II) is among the proposed candidate autoantigens initiaiting rheumatoid arthritis (RA). Statins have been shown to have anti-inflammatory and immunomodulating properties. Mechanisms of statin effects upon the autoantigen-induced T lymphocyte activation in RA patients are still unknown. The aim of present study was to evaluate the effects of mevastatin upon Col II- and anti-CD3-induced (PBMC) activation of peripheral blood mononuclear cells (PBMC) from RA patients. PBMC from active RA patients (DAS28 – 6.6±0.64) were stimulated with antibodies specific for either anti-CD3 or anti-Col II. The cultured cells were treated with mevastatin at different concentrations. Mevalonic acid, geranylgeranyl pyrophosphate (GGpp) and farnesyl pyrophosphate (Fpp) were added to cultures. Mevastatin at concentration of 10 μM caused significant reduction of Col II-induced PBMC proliferation and of IFNγ production. These mevastatin effects were partially reversible by addition of mevalonic acid, GGpp, and Fpp. Mevastatin did not influence the anti-CD3 induced PBMC activation. In conclusion, mevastatin is able to suppress antigen-specific T-cell stimulation in RA patients, due to decreased production of mevalonic acid metabolites.

Abstract. The work presents results of experimental study performed in 58 rabbits, using comparative evaluation of reparative events and dynamics of IgG values in experimental liver wound after surgical autoplastics with a serous muscular rag from the big curvature of stomach (45 animals), or common suture (13 animals). Terms of observation were from one day till 360 days. It is established, that studied autotransplant exerts additional transport of IgG into the liver wound, thus evacuating products of wound lesion and ischemic metabolism, providing a productive type of inflammatory process with adequate regeneration. After performing a liver suture, a trend towards chronic inflammation is noted since the earliest terms, followed by significant outgrowth of connective tissue within liver wound area at longer terms.

Abstract. Present article deals with our results concerning brain-specific S-100B protein levels, anti-S-100B autoantibodies of IgM and IgG classes, like as cytokine profiles of blood serum and cerebrospinal fluid in the patients with alcohol-induced delirium state. The results obtained provide an evidence of association between alcoholic psychosis and destruction of brain tissue, development of autoimmune reactions and altered cytokine status, thus, probably, resulting into disintegration of immune and neuroendocrine systems.

Abstract. Cytokines provide important connecting links between immunity, blood clotting, and nonspecific resistance that become altered in local frostbites. The aim of study was to determine the contents of cytokines in blood of patients with local frostbites at various terms following the lesion. Fifty patients, 17 to 50 years old, with frostbites of extremities (grade II to IV) were under observation. Arterial and venous blood, as well as venous blood from damaged and intact extremities were examined. Cytokine concentrations were determined using ELISA technique. It was shown that blood concentrations of TNFα, IL-1β, IL-4, IL-8 and IL-18 reached their maximum at early reactive period of the trauma (a 2.3- to 19-fold increase). IL-1β concentration exceeded appropriate control values during all the periods of local frostbites. IL 18 levels were increased at early reactive period of the trauma. IL-1β and IL-8 concentration at pre-reactive period proved to be higher in arterial blood of damaged extremities, than in effluent venous blood. Both during early and late reactive periods, the levels of pro-inflammatory cytokines in effluent blood from damaged extremities were 1.2- to 8-fold higher than in venous blood from intact extremities, whereas IL-4 levels were 2- to 5-fold lower.

Abstract. Progressive obstruction and lung tissue remodeling comprise an important feature of the airways in COPD patients. The main processes involved in tissue remodeling in COPD are protease/antiprotease, oxidant/antioxidant imbalances, like as inflammatory and fibrotic events that contribute to development or progression of disease. TGFβ is a multifunctional growth factor that regulates synthesis of extracellular matrix proteins, primarily collagen and fibronectin, thus inducing fibrosis of respiratory ways. The aim of our study was to determine levels of TGFβ in serum and bronchoalveolar lavage fluid (BALF) of COPD patients. All the patients with COPD had increased levels of TGFβ in serum, as compared with subjects without COPD (p < 0.01), but there was no difference in TGFβ concentration between patients at different stages of disease. Increased phagocytic activity of blood monocytes was found in 81% of COPD patients, as compared to controls. Phagocytosis of apoptotic T­cells and bacterial infection of monocytes leads to increased secretion of TGFβ and it may cause higher levels of TGF β in peripheral blood. TGFβ concentration in BALF of patients at stage III of disease was higher than in the patients at stage II (p < 0.05). The level of TGFβ in BALF directly correlates with number of alveolar macrophages (r = 0.39; р = 0.03). These data indicate that TGFβ is involved in chemotaxis of macrophages in COPD patients’ airways. We conclude that increased secretion of TGFβ by peripheral blood monocytes may be a result of their high phagocytic activity. Hence, TGFβ mediates interactions between the two main components underlying lung tissue remodeling, i.e. fibrosis of respiratory airways, and development of emphysema in COPD.