Abstract. The MHC molecules are the strongest transplantational antigens determining direct intraction with 95-98% of T cell clones activating in response to allograft. Interaction with MHC molecules is critically important for positive selection of thymocytes and subsequent acquiring the capability to recognize "self" and "foreign". In this light, alloreactivity was recognized as cross-reactivity with foreign MHC that appeared as result of repertoire selection on "self" MHC molecules. The T lymphocyte repertoire is formed in the thymus as a result of random rearrangement of germinal sequences of TCR gene fragments and other processes that bring about the diversity of TCRs. As shown by D. Raulet and co-workers, repertoire of T lymphocytes artificially formed at the absence of selection processes, is inherently capable of reacting with different allelic forms of MHC molecules. In contrast to germinal sequences of TCR fragments, the MHC molecules are characterized by a significant interspecies polymorphism. So, negative and positive selection are aimed at the adaptation of repertoire to the specific MHC-environment of certain individual. The overall goal of adaptation is the elimination of autoreactive clones and sparing a broad spectrum of specificity to potential pathogens. Recent results have shown that positive selection in thymus was result of degenerative recognition of endog- enous MHC/peptide complexes. In contrast, central, telerance to "self" is formed as result of high affinity specific interaction with self MHC/peptide complexes. From this viewpoint the negative selection in the thymus can be considered as a life-long allogenic reaction of repertoire to self MHC molecules; the responses of the mature T cell repertoire to individual antigenic peptides can be considered as cross-reactions of T cells reactive to foreign transplantational antigens. I believe that this interpretation can unveil the origins and biological mean-ingfulness of allogenic reactions.

In this review, the reasons for successful transplantation of stem cells have been shidied. Special attention has been paid to the influence of some stem cells types on thr immune system, mainly, on activated T-lymphocytes. It has been shown, that mesenchymal and stromal stem cells in vitro blocked the production of T-lymphocytes activated by allogenic cells or non-specific mitogens. Thus, contact with stem cells can impede the development of the graft immunity and inhibit the development of new T-lymphocyte clones. After the immunosuppressive therapy and stem transplantation of CD4+ population, as well as the impediment of the restoration of T-lymphocyte repertoire are observed. This effect is probably connected with immunomodulatory (similar to immunoregulatory) capacity of transferred allogenic or autologous stem cells. These studies have shown the risk of immunosuppression connected with the application of stem cells.

Abstract. We demonstrated that TNFα, IFNγ and IL-4 induce different patterns of endothelial cells activation. The greatest alterations were observed in TNFα threated endothelial cells. Under the influence of this cytokine there was a statistically significant elevation of IL-8 and MCP-1 production by endothelial cells, of THP-1 adhesion and of migration toward endothelial cells monolayer. At the same time we had not observed any immunophenotype alteration of THP-1 cocultured with endothelial cells treated with TNFα.

IFNγ и IL-4 induced statistically significant inhibition of THP-1 adhesion. IL-4 treated endothelial cells have elevated levels of IL-8 and MCP-1 production and stimulated THP-1 migrations. IFNγ showed inhibition of IL-8 and MCP-1 spontaneous production and THP-1migration.

Incubation of THP-1 with untreated and IFNγ treated endothelial cells led to significant elevation of HLA-DR expression. Expression of CD68 was significantly higher in case of THP-1 cocultured with IL-4 treated endothelial cells. There was no any correlation between immunophenotype alteration THP-1cells and others parameters alteration.

While investigating the efficacy of an H5N2 ca reassortant vaccine candidate in protecting against a lethal challenge with a highly pathogenic (HP) H5N1 virus in the mouse model, we observed a degree of cross-protection provided by the ca Len/17 (H2N2) virus itself. 80% of mice administered a high dose of attenuated Len/17 vaccine intranasally (i.n.) survived after a lethal challenge with A/Hong Kong/483/97 H5N1 virus. Therefore, we investigated the basis of the cross- reactive immunity between H2N2 and H5N1 viruses that may have contributed to recovery from lethal HK/ 483 virus infection. Sera from mice immunized i.n. with Len/17 did not cross-react with HK/483 virus in neutralization or hemagglutination-inhibition assays, however IgG and IgA antibodies that cross-reacted with the hemagglutinin and neuraminidase of H5N1 1997 viruses were detected. Spleen cells from mice immunized i.n. with Len/17 vaccine showed enhanced production of IL-2, IL-4, IL-5, IL-10, and IFNγ following in vitro stimulation with inactivated H5N1 virus. Our findings indicate that both cross-reactive humoral and cellular immunity induced by Len/17 H2N2 vaccine may plays a role in recovery from lethal H5N1 virus infection. A better understanding of the mechanisms of heterosubtypic immunity will improve vaccine design against HP avian influenza viruses.

Abstract. The presence of anti-thyroglobulin antibodies (AbTg) and their epitope specificity were investigated in sera of healthy people and patients with autoimmune thyroiditis (before and after L-thyroxine treatment). In sera samples AbTg were detected. It was shown that: (i) the level of AbTg in patients with autoimmune thyroiditis was significantly higher as compared with healthy individuals. After 6 months the level of AbTg of treated patients decreased but was higher than in healthy people. In serum samples of patients without L-thyroxine treatment the level of AbTg did not change. (ii) In healthy individual serum samples AbTg to antigenic regions of Tg recognized by monoclonal antibodies used in the work were not found. In patients serum samples monoclonal antibodies directed to 1F10 epitope were detected. Moreover the presence of monoclonal antibodies of the current specificity correlated with the increased level of AbTg. In 6 months the number of patients with AbTg to 1F10 epitope increased both in L-thyroxine treated and control group of patients. It is possible that the presence of AbTg to 1F10 epitope is associated with the pathological process in thyroid gland.

Abstract. The goal of our research was comparative study of the most important parameters of subset cytoarchitectonics in the patients with the different courses of myocarditis and evaluation of their pathogenetic and clinical value in the practice of the physician. We have investigated 99 patients with myocarditis and 40 healthy donors. In patients with malignant course of disease we revealed increased activation index of T/B-cells; increased expression of the activation markers on the both lines of differentiation; disproportion in the immunoregulatory subsets with increased role of dendric cells; decreased intensity of the autoreactive T-cells apoptosis. in the patient with the In patients with nonmalignant course of disease expressed signs of immunopathology were not found. Thus, study of activation markers on the cells of the peripheral blood is more informative and noninvasive method of diagnostics of myocarditis.

Abstract. The effects of 'Profetal', a drug containing liophylized, dextran-stabilized human alpha-fetoprotein as main active component, were investigated on peripheral donor blood mononuclear leukocytes (ML), with respect to their functional properties and the potential to generate mature dendritic cells (DC).

Addition of the drug to cell cultures at optimal doses was shown to cause significant increase in their proliferative capacity and ML blastic transformation levels, and enhanced cytotoxicity towards K562 tumor cells, as well as to induce maturation of antigen-presenting dendritic cells.

On the basis of the data obtained, a conclusion is made that 'Profetal' is an active immunomodulatory factor, and it may be applied for generation of cytotoxic lymphocytes and mature DC, aiming to apply them for the biotherapy of oncological and infectious diseases.

Abstract. Uterine myoma is one of the most widespread gynecological pathology among reproductive women older than 30 years. It is known, that often progress of this pathology is associated with genetic and endocrinologic factors. The immune system is not evident still. The aim of this study was to analyze the state of patient's immune system using flow cytometry assessment of different subpopulations of lymphocytes in peripheral blood. We have examined 46 patients with simple and proliferating forms of the myoma, with different variants of clinical symptoms. Absolute and relative content of different subpopulations of lymphocytes was not differed from normal population's standard. Significant differences of B-lymphocytes and natural killers content were observed between groups with simple and proliferating forms of disease. It was shown that metrorrhagia is associated with high level of T-lymphocytes and T-killers. It was noted that decreasing of B-lymphocytes content took place in cases with large number of uterine nodes. Obtained data are not sufficient for complete understanding of the role of immune system in pathogenesis of this disease, but they confirm that using of immunomodulating therapy is expedient for complex treatment of uterine myoma.

Abstract. Human alpha–2–macroglobulin (α2–MG) acts as a broad–spectrum cytokine and proteasebinding protein, and it represents an evolutionarily conserved arm of innate immune system. Meanwhile, previous studies have shown that rheumatoid arthritis (RA) development may cause alterations in α2–MG conformation and lessen its ability to bind and utilise regulatory substances. We investigated serum contents of α2–MG–IgG and α2–MG–plasmin complexes, as well as total concentrations ofα2–MG, plasmin (Pl), IgG, IL–6, IL–1β, TNFα and Rf–IgM, in order to evaluate the levels of anti–α2–MG antibody production in sera of patients with different degrees of RA activity and some interrelations of preformed immune complex with some other substances implicated in RA development. Serum samples were obtained in acute phase of RA, before the treatment was started. We have revealed significantly increased levels of α2–MG–IgG complex in the groups with severe RA, accompanied by increase in total IgG levels, without significant changes in total α2–MG concentrations. We have also demonstrated that increased levels of Pl–α2–MG complex did correspond to the severity of disease, and showed statistically high correlation with α2–MG–IgG levels. We have found a significant increase of IL–1β, IL–6, TNFα and Rf–IgM in RA, in absence of significant correlations with α2–MG–IgG contents. The results obtained allow us to suggest that abundant accumulation of serum antibodies to α2–MG or Pl–α2–MG complex during inflammation in the people with innate α2–MG deficiency, followed by increased levels of proinflammatory cytokines, may provoke a cascade–like development of RA. Serum levels of α2–MG may serve as prognostic marker in RA. (Med. Immunol., 2005, vol.7, № 5–6, pp. 557–562)

Abstract. The study included patients with chronic uveitis caused by herpes simplex virus (HSV), or mixed HSV/ cytomegalovirus (CMV) infections. Deficiencies in CD3+ and CD4+ T lymphocytes and significant increase of CD8+ lymphocyte subpopulations were found in posterior uveitis. A single-pathogen (HSV) infection was characterized by decreased expression of CD3+ and CD4+ lymphocytes, IgG hypoglobulinaemia. Irrespectively of etiology, both groups of the patients exhibited sharp increase in CD16+ cells, as well as suppressed CD25+ receptors for IL-2, increased numbers of activated CD38+ lymphocytes, enhanced expression of late activation markers (CD95+ and HLA-DR+) in both mixed and single-pathogen uveitis. IFN-alpha is detected at low concentrations, or is absent; increased TNF-alpha production was noted in the tears in cases of posterior uveitis.

Statistically significant interrelations have been found between expression of HLA-DR+, and CD3+; CD95+ and HLA-DR+; CD95+ lymphocytes and TNFalpha levels in the course of chronic persistence of Herpesviridae in the patients with chronic uveitis. The degree of immunological disturbances correlated with severity of clinical features.

Abstract. The aim of the present work was to evaluate the contents of some lymphocyte sub-populations in peripheral blood of the children with tumors of hematopoietic and lymphoid tissues, depending on infectious complication of cytostatic therapy and neutropenia. In all children undergoing cytostatic therapy for acute lympho-blastic leukemia and non-B cell non-Hodgkinґs lymphomas, we found significant decrease in the numbers of CD95 lymphocytes, absolute amounts of natural killer cells (CD16, CD56-lymphocytes) and activated lymphocytes (СD11b, HLA-DR-cells), irrespective of neutrophile numbers in their blood and infectious complications. However, absolute number of CD25- lymphocytes was significantly decreased in the children with neutropenia. Relative contents of CD16, CD56, СD11b, HLA-DR, CD25-lymphocytes did not significantly differ from those in healthy children, or they were found to be significantly increased.

Abstract. Clinical and immunological investigation of 103 patients with postoperative ventral hernia including the detection of lymphocytes subpopulations, the levels of IL–4, IFNγ and clinical manifestation of immune deficiency was carried out. The patients with postoperative ventral hernia had various changes of cells immunity connected with obesity. The low level of cytokines in all patients and disbalans of Th1– and Th2–cells have beenrevealed. The analyses of revealed abnormalities in comparison the results of clinical investigations were carried out. The results of this work are recommended to take into account during preoperative and postoperative periods. (Med. Immunol., 2005, vol.7, № 5–6, pp. 563–568)

Abstract. The purpose of study was to compare phagocytic indexes, free oxygen radicals generation, activity of antioxidative protection, cytokine levels and von Willebrand factor (vWF) antigen in the patients with non–stable course of ischemic heart disease of. We examined 58 patients with ischemic heart disease: 31 with unstable angina and 27 with acute myocardial infarction. The methods of study included nitro–blue tetrasolium reduction test for neutrophiles and monocytes, detection of myeloperoxidase in phagocytes, assays of glutatione reductase in neutrophiles, as well as measurements of catalase, superoxide dismutase, malonic dialdehyde, circulating immune complexes, IL–6, TNFα and vWF factor antigen in blood. Upon admission to the hospital, the patients with unstable angina, as well as with acute myocardial infarction showed significantly higher levels of vWF, C–reactive protein, neutrophile myeloperoxidase and appropriate trends in circulating immune complexes, IL–6 and TNFα. In the patients admitted to the hospital with ischemic heart disease with repeated coronary events during the last year of follow–up, as well as in acute myocardial infarction, the levels of vWF antigen were found to be significantly increased. The raise of vWF was accompanied by increased levels of malonic dialdehyde, C3R–induced generation of free oxygen radicals in the neutrophils, and drop in glutatione reductase activity, thus reflecting an “oxidative stress” condition. These patients did also exhibit higher levels of C–reactive protein, circulating immune complexes, IL–6 and TNFα. According to regression analysis data, the levels of vWF antigen and C3R–induced activation free oxygen radicals in neutrophils are the indices that were most closely connected to ischemic heart failure disease within a year of observation. The results obtained point to the important role of “oxidative stress” in the development of endothelial dysfunction in the patients with non–stable course of ischemic heart disease. (Med. Immunol., 2005, vol.7, № 5–6, pp. 569–574)

Abstract. Generalized forms of meningococcal infection (GFMI) are usually accompanied with apparent symptoms of systemic inflammatory response (SIRS). Systemic production of inflammatory cytokines, their accumulation in circulating blood and action on various organs and tissues represent a crucial constituent of SIRS. Current study explored the levels of soluble receptor of tumor necrosis factor (TNF) p55, TNF, neutrophil-derived alpha-defensins (HNP) and both isoforms of Interleukin-8 (IL-872 and IL-877) in blood and cerebrospinal fluid (CSF) from children with GFMI. The levels of cytokines were estimated using "sandwich" enzyme-linked immunoassays emploing monoclonal antibodies to corresponding cytokines that were previously developed in our laboratory. It was shown that the levels of p55 in the blood correlated with severity of SIRS and GFMI, while the levels of HNP (human neut rophil peptides) in CSF may serve as more sensitive marker of inflammation since HNP levels decreased more rapidly during a recovery of disease comparing to p55. Elevated levels of IL-877 during GFMI correlated with the levels of p55 (correlation coefficient 0,142, р = 0,489), whereas no correlation between the levels of p55 and IL-872 was revealed.

Abstract. We explored the effects of defensins on IL-8 synthesis in various human cells, including bronchoepithelial cell line A-549, monocytes, monocyte-derived macrophages differentiated in the presence of M-CSF, HU-VEC cells, HEK293 and THP-1 cell lines. HNP at 10-5-10-4М induced IL-8 production and cytotoxicity in serum-less A-549 culture. The addition of serum abrogated the cytotoxicity along with the induction of IL-8 synthesis. Similar effects were observed in HEK293 cell line and HUVEC.

The induction of cytotoxicity along with IL-8 production by HNP at 10-5-10-4М was also observed in the serum-less cultures of human monocytes and macrophages. However, monocytes and macrophages retained HNP-induced production of IL-8 after the addition of serum, while cytotoxic effect of HNP was completely inhibited. The data imply a specific interaction of alpha-defensins with some membrane receptors of monocytes and macrophages that leads to the induction of IL-8 and might also occur in vivo.

Abstract. The course of infections caused by intracellular parasitic bacteria Salmonella typhimurium and Chlamidophila pneumoniae was compared for inbred strains of mice with genetically determined susceptibity (I/St) or resistance (A/Sn) to Mycobacterium tuberculosis.

Similar differences in dynamics of some common parameters (life span and pathogen numbers in affected or gans) between A/Sn and I/St mice have been revealed for tuberculosis, salmonellosis and chlamidiasis. There has been demonstrated that A/Sn animals show increased resistance to Salmonella and Chlamidia infections, as compared to I/St mice, in spite of various mechanisms of intracellular parasitism for S.typhimurium and C.Pneumoniae, and different locations of lesions induced by these bacteria in the host organism.

Similar features of quite different infections at the level of macroorganism allows us to suggest that further investigation in A/Sn and I/St murine model will make it able to discover the basic features of physiolog$ical control for a lot of infections, and to reveal a genetic network that could be responsible for their favorable or adverse outcomes. (Med. Immunol., 2005, vol.7, № 5–6, pp. 583–586)

Abstract. Parameters of peripheral blood lymphocytes subpopulations have been studies in the patients with extensive destructive pulmonary tuberculosis caused by drug-sensitive or drug-resistant Mycobacterium tuberculosis before and in the course of specific anti-tuberculosis chemotherapy. It was revealed that in the patients before treatment, a marked immunosuppression takes place, being characterized by decrease of total T-lymphocytes and their basic regulatory and effector subpopulations. E.e., an increase in CD20+-, CD25+- and CD95+cells was observed. The completion of intensive anti-tuberculosis chemotherapy is not accompanied by correction of lymphocyte subpopulations. By contrary, in the patients with drug-sensitive pulmonary tuberculosis M. tuberculosis a sharp increase in CD95+-lymphocytes is registered. After undergoing a full course of therapy a well-noted tendency to normalization of T-lymphocytes (CD3+) quantity, as well as is regulatory populations (CD4+ and CD8+) were found. The level of CD95+-lymphocytes remained increased in the patients with pulmonary tuberculosis caused by drug-resistant M. tuberculosis.

Abstract. The aim of the investigation was to study the immunological characteristics of RA patients with anaemia. Clinical and laboratory data including the percentage of the main lymphocyte subclasses, phagocyte and DTH-effector activity, serum concentration of immunoglobulins, the percentage of cells producing IFNγ and/or IL-4 and percent of monocytes producing TNF. We revealed some significant clinical, laboratory and immunological differences between RA patients and healthy donors and between patients with and without anaemia. Our data demonstrate RA anemic patients to have more severe disorders than patients without anaemia. We also revealed some significant immunological differences between RA patients and healthy donors and between patients with and without anaemia, including percent of cells producing IFNγ and/or IL-4. Our data permit to conclude that RA patients have many different immunological disturbances, more severe in anaemic patients.

Abstract. Study results of vaginal secretion cytokine system in 25 patients with urogenital trichomoniasis are presented. Concentration of interleukins:IL-1α, IL-1β, RAIL-1, TNFa, IL-8, IFN-γ has been performed. The character of pro-inflammatory cytokines changes in the local immunity system in females with urogenital trichomoniasis has been studied in comparison with these values in healthy females. It has been established that infected females have increased levels of IL-1α, IL-1β, RAIL-1, TNFα, IL-8, while IFN-γ concentration is decreased. The obtained results enable to get a deeper insight into the cytokine-mediated reactions in the local immunity system of genital tracts in case of trichomoniasis.

Abstract. Intraperitoneal injection of rIL-1β (1 mkg/kg once a day within 5 days) during proceeding fractional irradiations (0,5 Gy once in day, the beginning of therapy after the total doze 20 Gy) prevents the further decrease in quantity of leukocytes in peripheral blood and oppression of alkaline phosphatase activity in neutrophils of irradiated rats, and also accelerates their restoration after cessation of radiatijn exposure. Restoration of the general number of leukocytes, absolute quantity of neutrophils and lymphocytes, normalization of the alkaline phosphatase activity in neutrophils of rats after the end of fractional irradiations in a total doze 25 Gy at course administration of rIL-1β occur in earlier terms, than in the animals of the control group.

Abstract. Ulcer disease is accompanied by all immune responses with the development of secondary immune deficiency. We studied 92 males with peptic ulcer having positive tests for Helicobacter pylori (24 gastric ulcer, mean age 44,7±1,97 and 68 duodenal ulcer, mean age 31,5±1,71). A half of the patients used only basis antiulcer treatment. 46 patients used bestim in the dose100 mcg per day intramusculare for a 5 days with antisecretory and antihelicobacter drugs. The addition of bestim normalized cellular immunity: count of T-lymphocytes rose from 20,52±0,89% to 31,4±1,16% (p<0,05; confidence interval [CI]: 2,67-11,63). A positive trend in the number of T-lymphocytes was accompanied with increasing of T-helpers from 17,33±0,89% to 24,82±1,11% (p<0,05; 95%CI: 1,72-8,47). In the course of the treatment we observed normalization of monocytes number, an increase of neutrophils activity in blood (p<0,05; 95%CI: 2,35-15,64). We demonstrated decrease in IL-8 production and increase in IFN gamma production. Thus, the application of a new drug Bestim in the combined antiulcer treatment affects the immune state of the patients favourably. It decreases the inflammatory effect in the stomach and duodenal mucosa, it precipitate regress of clinical symptoms and healing of the ulcer.

Abstract. Salmon soft roe DNA influence on healthy donors blood cells secretion of early hemopoietic factors (IL-3, GM-CSF, TNFα) as well as biologically active substance influence on cytokine balance of Тh1 and Тh2 responses (IFNγ, IL-10) in vitro was studied. It is established, that DNA has modulatory effect on secretion of all investigated cytokines - IL-3, GM-CSF, TNFα, INFγ and IL-10 by blood cells of healthy donors, increases their initially low concentration, reduces initially high and does not have essential influence at an average level of their secretion. Under action of DNA IFNγ level (stimulation index=3,3) increases more significantly than IL-10 level (stimulation index =1,9). Thus, salmon soft roe DNA possesses immunomodulatory properties.

FOR THE MEMORY OF PROFESSOR VADIM GALAKTIONOV (1938–2005)