Precise mechanisms of tolerance loss to thyroid auto-antigens are still largely unknown, thus preventing further elucidation of key pathogenetic stages in autoimmune diseases affecting thyroid gland, and, therefore, to identify target pathways for treatment of these disorders. Detailed studies on the role of innate and adaptive immune cells in disturbed tolerance to autoantigens specific for thyroid gland in Graves’ disease and autoimmune Hashimoto’s thyroiditis have revealed and provided a number of theoretical foundations for arrangement of clinical immunological monitoring and forecasting of these diseases in a long-term prospective.

The aim of present study was to arrange an experimental rabbit model for IgA-nephropathy. To this purpose, an approach was attempted which was previously successfully applied in rabbits, aiming for induction of post-streptococcal glomerulonephritis (PSGN). To induce the nephropathy, we used two Streptococcus pyogenes strains of M4 and M60 serotypes which showed differential IgA-binding capacity mediated by the IgAFc microbial receptors. The renal tissue damage was developed in most animals treated with emm60 S. pyogenes characterized by marked IgAFcR expression and higher IgA-binding ability. By means of morphometric analysis. Significant morphological and immunochemical glomerular changes were revealed in 6/10 rabbits, as follows: (i) massive IgA deposition in the mesangial glomerular cells, atrophy of the capillary net, and tissue oedema; (ii) marked C3-complement deposition in proximal and distal tubules; (iii) a significant infiltration of cortical and medullar areas by lymphocytes associated with weak TNFα production. Noteworthy, we did not observe local IgG deposition in any cases, thus allowing to exclude any role of anti-IgG, or other IgG’s in evolution of the pathology. Alternatively, the IgA-deposits may occur due to microbial IgA FcR–IgAcontaining cоmplexes, as earlier shown by the Swedish scientists. The above tissue changes were completely absent in kidneys of control animals. Taken together, these data suggest that we have developed an experimental model similar to IgA-nephropathy in humans. The results also extend our knowledge on pathogenic effects of the IgA Fc-binding proteins of Streptococcus pyogenes.

The purpose of this study was to assess some mechanisms of changes in immune state, and to evaluate a role of amlodipine, a known calcium channel blocker, as a potential corrective drug in experimental chronic renal failure (CRF). An animal CRF model was produced in rats by a two-stage operative resection of 5/6 of the renal tissue. Amlodipine is used per os at a daily dose of 0.25 mg/kg for 7 days. Flow cytofluorimetric approach was used to discern peripheral blood lymphocytes: CD3+ (mainly, T lymphocytes), CD45RA+ (mainly, B cells), as well as the following cell markers: Annexin 5-FITC+/7-AAD- (early apoptosis), Annexin 5-FITC+/7-AAD+ (late apoptosis and, in part, necrotic cells). Moreover, we have measured serum concentrations of urea, creatinine, phosphate, total calcium, parathyroid hormone (PTH), IL-1β, IL-4, interferon-γ, superoxide dismutase (SOD) and catalase activities. Evaluation of Th1- and Th2-dependent immune response was carried out, respectively, by detection of delayed-type hypersensitivity, and scoring the antibody-forming cells in rat spleen induced by immunization with allogeneic erythrocytes. Primary, secondary and final products of lipid peroxidation were evaluated in lipid extracts from peripheral blood lymphocytes. Changes of immune state in CRF included depression of Th1 and Th2 dependent immune response, reduced number of lymphocytes bearing T and В cell markers, increased IL-1β concentrations in blood, along with decreased amounts of IFNγ and IL-4. Probable pathogenesis of the altered immune state may be associated with increased number of peripheral lymphocytes being at early and late stages of apoptosis/necrosis, elevated blood levels of IL-1β, total calcium, parathyroid hormone, reduced concentrations of IFNγ, and increased contents of primary, secondary and final peroxidation products in peripheral blood lymphocytes, being accompanied by inhibition of the SOD and catalase activity in blood plasma. Amlodipine administration lead to restoration of Th1 and Th2 dependent immune response, increased number of cells positive for T and B lymphocyte markers. Probable immune mechanisms of amlodipine effects may be associated with decreased numbers of apoptotic/necrotic lymphocytes, reduced lipid peroxidation in peripheral blood lymphocytes, enhanced SOD activity in blood plasma.

Expression of chemokine receptors (CCR4, CCR6, CXCR3 and CXCR5) on T-helper (Th) cells at various levels of differentiation in a group of healthy volunteers (n = 52) was assessed on the basis of CD45RA and CD62L expression, using the eight-color flow cytometry. It was found that the “naive” T helper cells (N) with CD45RA+CD62L+ phenotype express CXCR3 (4.94±0.39%), and CXCR5 (3.63±0.25%). About 50% of central memory T helpers (CD45RA–CD62L+, CM) were CXCR3 positive, and 43.72±1.27% of CM cells expressed CCR6, whereas CXCR5 and CCR4 levels were about 30%. Furthermore, CXCR3 was expressed by 76.76±0.75% of the CD3+CD4+CD45RA–CD62L– (EM) population, and similar values were obtained for CCR6, while the relative abundance of CXCR5+ cells decreased to 13.68±0.50%, and CCR4 levels did not change and accounted for 33.26±1.13% positive cells. Likewise, co-expression of the chemokine receptors was studied for the abovementioned subpopulations of T helper cells. Among the CXCR5– Th, Th1 cells were identified as CXCR3+CCR6–CCR4– (this subset also contained Th9), and CXCR3+CCR6+CCR4– subsets, referred to as Th1/Th17. Th2 were detected on the basis of CCR4 expression in absence of all other chemokine receptors. In addition to the mentioned Th1/Th17 populations, Th 17 cells were found in the subsets of Th17 CXCR3–CCR6+CCR4– and CXCR3–CR6+CCR4+. 
The latter also contained a Th22 population. Follicular Th cell populations (CXCR5+) consisted of, at least, six different subsets: CXCR3–CCR6–CCR4– (Tfh/Tfh2), CXCR3–CCR6–CCR4+ (Tfh2), CXCR3-CCR6+CCR4–(Tfh17), CXCR3–CCR6+CCR4+ (Tfh17), CXCR3+CCR6–CCR4– (Tfh1) and CXCR3+CCR6+CCR4–(Tfh1/Tfh17). The cells with Th1/Th9 and Th1/Th17 phenotypes dominated among CM (about 13%), whereas their relative abundance within EM increased to 22.37±1.69% and 31.69±1.52%, respectively. The amounts of Th2 were 8.15±0.46% within CM, and only 1.72±0.15% for EM population. For the cells with Th17/Th22 phenotype, these values are 8.07±0.30% and 12.03±0.57%, respectively. The main Tfh subsets were represented among the CM T-helpers: the relative content of Tfh/Tfh2 was 5.79 0.26%, Tfh2, 1.34±0.07%; Tfh17 with CXCR3-CCR6+CCR4– and CXCR3-CCR6+CCR4+ phenotypes made up to 6.22±0.28% and 3.28±0.16%, as well as Tfh1 (7.68±0.31%), and Tfh1/Tfh17 (4.02±0.17%), respectively. Relative content of the mentioned Tfh subsets was decreased > 2-fold within effector memory Th subpopulation. The data obtained may be applied for diagnostics of different immunopathological conditions and could be used as a comparison group in further studies.

The aim of present study was to investigate relationships between cytokine-producing potential of immunocompetent blood cells (ICC), and expression of heparanase-1 in stomach adenomas and adenocarcinomas. Cytokine-producing potential of ICC, both spontaneous and induced by polyclonal activators, was studied in peripheral blood of thirty-two patients with adenomas and thirty-six patients with gastric adenocarcinoma. Appropriate values were expressed as an influence index of polyclonal activators (IIPA) upon the cytokine production. In subjects with gastric adenomas, the IIPA indexes based on IL-1ra, ТNFα, IL- 18, and IFNγ production proved to exceed similar parameters for the patients with adenocarcinomas. Expression of heparanase-1 in epithelium of adenomas varied from 0.27 to 3.00. Similar values for adenomatous stroma varied between 2.27 and 11.48. Meanwhile, the variation limits for heparanase-1 expression in adenocarcinomas (as percentage of specifically stained area) were quite considerable (1.79 to 14.26), due to inclusion of adenocarcinomas at variable differentiation stages. Indexes of heparanase-1 expression were shown to be in direct correlation with numbers of regional lymph node metastases. Multivariate analysis revealed a canonical correlation between the levels of heparanase-1 expression in adenomatous epithelium, and IIPA effects upon production of IL-2, IL-6 and IL-8, as well as with effects of IIPA upon production of IL-18, IL-18BP, and IL-8. A canonical correlation was also noted between the heparanase-1 expression in stroma of adenomas, and IIPA upon IL-1ra, TNFα, IL-18 production. The data suggest an involvement of molecular and cellular mechanisms, mediated by cytokines and heparanase-1, thus providing a common vector promoting tumor progression.

The aim of our study was to investigate the relationship between phenotype and «respiratory burst» activity of neutrophilic granulocytes in patients with renal cancer. The study included 73 patients with renal cell carcinoma (T3N0M0, clear-cell type) before surgical treatment at the age of 40-55 years. The diagnosis of renal cancer was histologically verified in all the patients. Fifty healthy age-matched persons were involved as a control group. Phenotyping of blood neutrophilic granulocytes was performed by flow cytometry. The respiratory burst in blood neutrophilic granulocytes was evaluated using spontaneous and zymosan-induced luminol- and lucigenin-dependent chemiluminescence. Some changes in relationships between the phenotypic indicators of neutrophils and «respiratory burst» were detected in patients with renal cancer. The patients exhibited a loss of relationship between induction of primary and secondary reactive oxygen species, and numbers of CD11b+, CD62L+, and CD64+ neutrophils. These correlations are detected with HLA-DR+ cell levels, like in control group. The patients’ samples showed a correlation between the superoxide radical synthesis by resting neutrophils, and CD62L expression. These relationships disappear when assessing antigen-induced «respiratory burst» response. The correlations between primary and secondary reactive oxygen species, and the numbers of 
CD11b+, CD62L+ and CD64+ neutrophils found in healthy people characterize a positive relationship between the «respiratory burst» indexes and activation of cells as reflected by expression of adhesive and functional receptors. It may be assumed that altered relationship between the «respiratory burst» indicators and neutrophil phenotype in the patients with renal cell carcinoma is determined by various factors. Firstly, cancer-associated phenotypic changes of blood neutrophils (decreased content of CD11b+, CD62L+, CD64+, HLA-DR+, and HLA-DR+ CD64+ neutrophils, increased expression of CD16, and HLA-DR receptors) thus, possibly, being connected with intra-tumor migration of the cells. Secondly, the patients show an increase in «respiratory burst» intensity in neutrophils, due to enhanced synthesis of primary and secondary reactive oxygen species, which may be connected to the action of various tumor factors (antigens, regulatory molecules).
The relationships revealed between phenotype and «respiratory burst» of neutrophils may specifically characterize immunopathogenesis of kidney cancer and can be used in development of immunotherapeutic approaches aiming to stimulate anti-tumor activity of phagocytic cells.

Mechanical jaundice (MJ) is a severe pathological condition, caused by obstruction of the bile ducts, requiring immediate surgical intervention. Etiologically, MJ can be of benign (60-80% of the cases,) or malignant origin. MJ progression depends on the underlying pathology, and, moreover, on bilirubin levels. Focal inflammation in affected area represents a significant mechanism of the MJ progression. Neutrophilic granulocytes, are primarily involved into the immune response, i.e., pathogen elimination. Hence, the MJ progression may depend on their functional activity. In this context, the aim of our study was to investigate chemiluminescent activity of neutrophil granulocytes in progression of MJ, depending on the bilirubin levels and origin of the jaundice. All the MJ patients showed altered chemiluminescent activity of granulocytes. Both spontaneous and induced chemiluminescence (CL) intensity was decreased in the patients with gallstoneassociated MJ. Meanwhile, the CL intensity did not change in MJ caused by benign tumors (BTP). An increased activation index reflected higher induced activity of phagocytes. In patients with MJ of malignant origin, the largest number of changes was found, i.e., an increase in the induced luminescence intensity and higher activation indexes were revealed. The MJ progression depends on blood bilirubin levels. All the patients with gallstone-related MJ exhibited a decrease in spontaneous and induced CL activity of the neutrophils. In cases of BTP-caused MJ, the indexes of spontaneous and induced CL decreased at the bilirubin levels of 60 to 200 mmol/L, with increased activation index suggesting elevated induced activity over its spontaneous levels. In patients with MJ and bilirubin levels >200 mmol/L, distinct unidirectional changes in the granulocyte activation were observed, with increased spontaneous and induced CL intensity. In patients with MJ caused by malignancies with bilirubin levels <60 mmol /L , both spontaneous and induced CL activity were reduced, whereas, at 60 to 200 mmol/L, a recovery of granulocytic activity was observed. In the cases of bilirubin levels of >200 mmol/L, an increased spontaneous and induced activity of granulocytes was registered in these cases, probably, due to development of multi-organ failure and toxic effects of bilirubin. The granulocytes seem to exhibit their maximal activity, until exhaustion of their functional limits.

CXCL10 (IP-10) chemokine is one of additional humoral markers applied along with determination of antigen-induced IFNγ for diagnosis of latent tuberculosis infection in humans. Dynamics of the IP-10 changes and its prognostic value in planning treatment for various forms of tuberculosis is a scarcely studied problem, especially, in children and adolescents. Present study has revealed an increase in spontaneous and antigen-induced IP-10 production in children and adolescents with overt tuberculosis, when compared with latent course of tuberculosis, followed by its significant reduction in cases of successful chemotherapy of tuberculosis. Such dynamics was not evident in destructive forms of tuberculosis manifesting as torpid and progressive infection.

We studied cytokine profiles relevant to the Th1 and Th17 modes of immune response in 116 patients with mild, moderate and severe chronic obstructive pulmonary disease (COPD). Relative contents of Th1- and Th17-type lymphocytes were evaluated by serum levels of specific cytokines: TNFα, IL-6, IL-10,IL-17A, IL-21, IFNγ, TGF-β1. We found that the shift in balance of pro- and anti-inflammatory cytokines is typical to both Th1 and Th17 immune response types in COPD of different severity. The variety of immune response types was achieved due to activation of different effector cells. I.e., the Th1 response was associated with macrophage activation and IFNγ induction, whereas domination of Th17 response was accompanied by activation of neutrophils, and increased synthesis of IL-17, IL-21. The Th17 type of immune response was shown to prevail in severe COPD. Thus, the certain types of immune disorders in COPD presume application of combined therapy including immunocorrective drugs which are able to suppress inflammation and correct the cytokine imbalances.

The aim of our study was to identify phenotypic composition and functional activity of immune cells from umbilical cord blood (UCB) as a function of gestational age. We analyzed 102 UCB samples of babies born at 25-28 weeks, 29-32 weeks, and 33-36 weeks of gestation, and 31 samples of UCB from the full-term infants taken as a comparison group (gestational age of 37 to 41 weeks). Using flow cytometry approach, we determined representation of cells with CD3+-, CD19+/CD3-, CD4+/CD3+, CD8+/CD3+, CD16/56+/CD3-phenotypes, along with expression of activation markers among populations of monocytes and lymphocytes (HLA-DR+/CD14+, CD71+/CD14+, CD25+/CD4+, CD95+/CD3+), as well as adhesion receptors (CD11b+). Contents of cytokine-producing cells (IL-4/CD4, IFNγ/CD4) and serum IFNγ and IL-4 cytokines were also determined. All premature babies with existing leukopenia exhibited an increase in relative numbers of lymphocytes, and diminished ability of the granulocytes to absorb opsonized bacteria. Reduced percentage of HLA-DR+/CD14+ cells, increase in CD3+CD95+ cell population and IFNγ producing cells have been shown in a subgroup of children at a gestational age of 25-28 weeks. Decreased levels of natural killer cells was observed among infants < 32 weeks of gestation. Increased concentration of serum IFNγ, along with reduced levels of IL-4 was determined in children born at 25 to 28 and 33 to 36 weeks. A reduced percentage of CD3+CD11b+ cells were revealed in UCB at 29-32 and 33-36 weeks of gestation. Reduced numbers of CD3+CD95+lymphocytes and increased CD25+ receptor expression on the CD4+ cell population was registered at 33 to 36 weeks of gestation. In summary, the findings suggest some features of immune system, which may be specific to particular gestational age of preterm infants. A number of the parameters under study allow us to presume a development of partial immunological response to potential antigens, thus requiring further research aimed for investigation of functional activity of immunocompetent cells.