We have studied dose effects of several Interferon (IFN) inducers, i.e., Genfaxon (beta-1 IFN), Cycloferon and Immunomax upon expression of six genes controlling the signaling in immune pathways (TLR3, TLR4, RIG1, IRF3, IPS, B2M), by means of real-time RT-PCR, being tested with blood cells from three humans. It is revealed that individual cell samples showed different sensitivity to these drugs, probably, due to constitutive levels of TLR3 and TLR4 gene expression and possible connections with their immune pathology. Genfaxon at a dose of 104 ME produced potent stimulation of TLR3, TLR4, IRF3 and B2M genes in two persons. Immunomax, at a dose 0,5 unit, exhibited same effect in one case only (with Epstein-Barr virus infection). Cycloferon stimulated gene expression at much lower levels than Genfaxon in any cases. We have shown a reverse correlation between sensitivity of the cells to Immunomax, and constitutive TLR3 and TLR4 expression. The stimulatory effects of Immunomax were maximal in a person with very low TLR3/4 gene expression. Immunomax boosted the genes from several signaling pathways, including TLR3, TLR4, but genes of RIG/IPS pathway showed higher activation. Cycloferon induced gene transcription of IRF3 and B2M-receptor to higher degree, than expression of TLR3 and TLR4 genes. Hence, our data concerning Genfaxon, Immunomax and Cycloferon confirm their IFN-inducing effects upon human blood cells. The RT-PCR-based evaluation of gene expression related to signaling immune pathways in blood cell populations will enable rapid and highly specific quantitation of IFN and IFN-inducer drugs activities, thus avoiding their biological testing in long-term cell cultures. 

The article deals with applications of six-color flow analysis for studying the immune state parameters by means of flow cytometry. Whole blood from healthy donors was stained with combination of monoclonal antibodies, i.e., HLA-DR-FITC, CD16+56-PE, CD4-ECD, CD19-ECD, CD8-PC5.5, CD3-PC7, CD45-APC (Beckman Coulter, USA) using a “no-wash” technology. To adjust the photomultiplier (PMT) voltage, we used the tubes with each of the tested monoclonal antibodies, PMT voltage was considered optimal when the negative population was located in the middle of the first decade at the logarithmic scale. The compensatory adjustment was performed in automatic mode, using Navios software (Beckman Coulter, USA). We discuss an optimal gating strategy in order to assess the populations of interest: total T cells (CD3+CD19-), T helper cells (CD3+CD4+), cytotoxic T lymphocytes (CD3+CD8+), B cells (CD3-CD19+), NK cells (CD3-CD16+CD56+), NKT cells (CD3+CD16+CD56+), activated T lymphocytes (CD3+HLA-DR+). 

The aim of this study was to identify and evaluate the features of immune response for main subpopulations of peripheral blood lymphocytes during exacerbation of chronic gastritis, depending on the severity of gastric mucosal atrophy. A comprehensive survey of 122 patients with acute exacerbation of chronic gastritis was performed. The patients with chronic focal-atrophic and atrophic gastritis in acute stage were characterized by increased numbers of CD3+ lymphocytes, CD4+, CD8+ and CD16+ cell populations, decreased CD19+ lymphocyte counts in peripheral blood. During exacerbation of non-atrophic gastritis, an increase of CD4+ and CD16+ cell counts was associated with a decrease in the CD8+ and CD19+ lymphocytes. A concomitant increase in CD4+ cell numbers, as well as elevated IL-2 and IFNγ levels in peripheral blood of patients may reflect activation of cellular immunity, which could be induced by the dysplasia of gastric mucosa and co-infection with Helicobacter/herpesvirus. Domination of increased IL-2 and IFNγ over IL-4 and IL-10 levels suggests the Th1-profile of cellular immune response. Upon exacerbation of non-atrophic gastritis, the immunoregulatory index exceeded the values of clinically healthy subjects and those of patients with atrophic gastritis. We observed statistically significant differences for CD3+, CD4+, CD8+, CD19+ lymphocytes, immunoregulatory index, and serum levels of IL-2, depending on presence and severity of gastric mucosal atrophy. 

Formation of the earliest forms of chronic brain ischemia veterans of modern wars accompanied by an increase in the systemic circulation of the population of T lymphocytes and monocytes, reflecting the activation of central mechanisms lymphopoiesis. In step vascular encephalopathy is an increase in circulating pool of T lymphocytes expressing the activation markers early positive reflecting readiness cells to IL-2 dependent proliferation. When progessirovanii chronic brain ischemia decreased levels of circulating T-regulatory cells, which may reflect a violation of self-tolerance in relation to brain antigens.

Some features of peripheral bood basophil activation were studied in children with different types of chronic urticaria, in order to specify the sustaining mechanisms of chronic urticaria. This study presents comparative data concerning spontaneous basophil activation, IgE level and Th2 helper cells. We observed forty-seven children with chronic urticaria, seventy children with exacerbated atopic dermatitis with different disease severity, fifteen healthy children without signs of allergy, or pseudo-allergy. High degree of the basophil activation was detected in children with exacerbation of atopic dermatitis (a comparison group), which was different from the groups of healthy children and patients with chronic urticaria. Spontaneous basophile activation was higher in children with chronic urticaria than in healthy children, as influenced by immune and non-immune pathogenetic factors of the disease. A positive pairwise correlation was shown between total IgE level, and Th2 helper cell numbers in peripheral blood in patients with chronic urticaria, thus confirming a Th2 immune shift. The results obtained contribute to our understanding of immune pathology in chronic urticaria. 

Certain correlations between changes of cytokine levels and kidney dysfunction, as well as their prognostic significance for development of acute kidney injury (АКI) in different clinical forms of ischemic heart disease (IHD) seem to be worth of further studies. The levels of IL-6 and IL-10 and their correlation with serum creatinine (sCr) and lipocalin levels in urine (u-NGAL) were studied in 98 IHD patients, of them 93 presented with acute coronary syndrome (ACS) and 5, with stable angina pectoris. ACS patients were found to have increased levels of IL-6, IL-10, sCr, u-NGAL. IL-6 and IL-10 contents proved to be increased in cases of non-ST-segment elevation ACS (NSTE-ACS), and during AKI development. According to serum creatinin levels, AKI was more frequently revealed in patients with NSTE-ACS. Urinary NGAL levels were found to be higher in ACS with ST segment elevation. 

We had investigated levels of TTG, T4, TNFα, IL-6, IFNγ, and α2-MG in blood serum and supernates of short-term blood cultures in the patients with verified Graves disease before treatment and after reaching of euthyroid status, as compared with healthy controls. We have revealed that initial blood concentrations of free Т4 in the patients were increased, along with decrease in TSH, higher IL-6, IFNγ levels, as well as concentrations of α2-MG which participates in cytokine transport and synthesis. Thiamazole treatment normalized the hormonal profile and reduced blood levels of IL-6, IFNγ and α2-MG, however, without complete normalization, along with increase of serum TNFα contents. It was shown, that the patients before treatment had decreased in vitro response of cells to the mitogenic stimulation as shown by decreased induction of TNFα and IFNγ production, along with, increased spontaneous IFNγ levels. When reaching euthyroid state after Thiamazole administration, we observed an increased spontaneous IFNγ synthesis, decreased IL-6 production in resting cultures. In mitogen-stimulated cell cultures from the treated patients, IFNγ contents became normal, however, TNFα secretion remained lower than in controls. The α2-MG levels in supernates were stable and significantly lower, than in serum. We may presume that thyrotoxicosis treatment with Thiamazole causes stabilization of the endocrine state, however, being not sufficient for normalized production of cytokines, as well as α2-MG, with its regulatory and transporter functions, thus promoting recurrence of disease and reactivation of autoimmune events. 

From the beginning of 21th century outbreaks of H5, H7 and H9 avian flu are registered from time to time. These viruses are considered as one of the possible causes of the next pandemia. The development of avian influenza vaccines is one of the WHO priorities. The aim of this work was to study antibody and cellular immune responses to avian A (H5N2) and A (H7N3) live attenuated influenza vaccines (LAIVs). We examined serum antibodies (HAI assay, microneutralization assay, ELISA), local antibodies (ELISA) and virus-specific CD4+ and CD8+ central memory and effector memory T cells. Two doses vaccination of healthy volunteers with A (H5N2) and A (H7N3) LAIVs induced homological antibody and cellular immune responses (i. e. serum and local antibody conversions, virus-specific memory T cell growth). These vaccines also stimulated heterological immunity (heterological serum and local antibodies and T cells). Heterological immune response intensity depended on antigenic structure of vaccine strain and heterological virus, particularly on HA type. 

The aim of this study was to evaluate a possible role of AID (AICDA – activation-induced (cytidine) deaminase) in the bronchial asthma (BA) pathogenesis. Materials and methods. We have examined twelve healthy control persons, forty-two patients with allergic bronchial asthma (ABA) and twenty-seven patients with non-allergic bronchial asthma (NABA). The AID mRNA expression was evaluated by means of RT-PCR. Results: AID mRNA was more significantly expressed in BA, than in healthy controls. Meanwhile, no significant differences were revealed between ABA and NABA groups. Correlation analysis of AID mRNA expression levels in peripheral blood lymphocytes has shown that CHε mRNA and total serum IgE revealed significant negative correlation, in the NABA group only. The levels of AID mRNA expression in peripheral blood lymphocytes exhibited positive and significant correlations with clinical characteristics, reflecting severity and stage of the disease, in BA patients. Moreover, a significant positive correlation was revealed with eosinophil contents in sputum. Conclusion. It was concluded that normal regulation of IgE class switching normally based on feedback regulation, was impaired in ABA but not affected in NABA. 

The study included 90 patients with pulmonary tuberculosis who were divided into 3 groups, according to intensity and clinical form of the disease. Group 1 included 32 patients with infiltrative tuberculosis and two-segment pulmonary lesions; group 2, 31 patients with pulmonary tuberculoma, and group 3, 27 patients with fibrous/cavernous tuberculosis for less than two years after the process stabilization. Group 4 represented a control (comparison) group of 30 healthy volunteers. The analysis showed that amino acid balance of monocytes in pulmonary tuberculosis can be, in general, characterized as deficient for antioxidant resources. In case of tuberculous inflammation, the criteria of specific resistance comprise a necessary taurine pool to greater extent than glutathione oxidation in immunocompetent cells. The plasma/monocyte taurine ratios exhibit different values, depending on clinical form of pulmonary tuberculosis. Redistribution of plasma amino acids to the monocytes, or, vice versa, probable “washout” certain amino acids from the cells may be considered a factor that reflects severity of the infectious process. 

Sixty-eight patients with acute coronary syndrome (ACS) were studied for correlations between the levels of interleukin IL-6 and IL-10 in blood and saliva, highly sensitive C-reactive protein (hs-CRP) and brain natriuretic peptide (NtproBNP) in the blood serum with the development of cardiovascular complications (CVC) during hospital period of the disease. It has been revealed that the patients with CVC had higher concentration of IL-6 in blood, IL-6, IL-10 in saliva, hs-CRP and NtproBNP in blood samples. Meanwhile, excess of IL-6 levels in saliva over those in blood was a significant predictor of CVC development. In order to facilitate the prediction values of CVC during ACS hospital period we have proposed a “CVCACS” model that employed the parameters of patient’s age, IL-10 level in the saliva, IL-6, and hs-CRP amounts in blood. It should be mentioned that the variable value of > -0,657 obtained with “CVCACS” model was predictive for development of CVC during ACS hospital period, while ≤ -0,657 was associated with favorable course of ACS hospital period. 

Osteoarthritis (OA) has been shown to be a heterogeneous disease. Diabetes mellitus (DM)associated represents a special OA subtype. Its clinical and immunological characteristics are poorly understood. To assess immune phenotype of the diabetes-associated OA and appropriate relationship between its clinical manifestations and cytokine concentrations in peripheral blood, we examined 78 patients with generalized OA including 52 patients in experimental group (82.6% females) who exhibited clinical manifestations of OA preceded by DM type II for, at least, 1 year, and 26 OA diabetes-free patients (84.6% females). We found that clinical manifestations of DM-associated OA were associated with increased body weight, more pronounced level of joint pains, longer duration of morning stiffness, decreased functionality of hands and large joints, impaired quality of life and more severe clinical pattern of the illness. Pronounced clinical manifestations in OA patients were more typical to the patients who required insulin therapy. The patients with DM type II-associated OA had elevated levels of proinflammatory (IL-6, IL-18) and reduced serum concentrations of anti-infammatory cytokines (IL-10, adiponectin), thus suggesting more pronounced systemic inflammation in patients of the first group. Concentrations of circulating IL-6 correlated with several functional indexes of OA severity. In conclusion, the DM-associated OA represents a special subtype of osteoarthritis, and deserves further studies of its immune pathogenesis and development of new treatment strategies. 

We are studied the 15 patients with autoimmune liver diseases and 36 patients without autoimmune pathology found the diagnostic value of antinuclear and antimitochondrial autoantibodies (AMA-M2) tests, and antibodies to asialoglycoprotein receptor (anti-ASGPR). Based on the ROC analysis showed that the diagnostic sensitivity and diagnostic specificity of AMA-M2 was 73% and 100% and for anti-ASGPR – 60% and 77%, respectively. Therefore, the test for anti-ASGPR in autoimmune diseases of the liver showed no advantages over standart tests, and its using in clinical practice requires clarification.