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Type 2 diabetes mellitus is characterized by a mild inflammatory reaction in the pancreas, which affects the structure and function of the pancreatic islets: the number of β-cells decreases and the number of α-cells increases. The work examined the features of β-cell differentiation in the development of experimental type 2 diabetes mellitus and while reducing the inflammatory process. Biochemical, histological methods, enzyme-linked immunosorbent assay, immunohistochemical methods were used using primary antibodies to insulin, glucagon, proliferation marker Ki-67 and secondary antibodies labeled with fluorescent dyes. Streptozotocin and nicotinamide were used to model type 2 diabetes mellitus, and the sodium salt of 5-amino-2,3-dihydrophthalazine-1,4-dione was used to reduce the inflammatory response. Previous studies have shown that it changes the macrophage phenotype from proinflammatory M1 to anti-inflammatory M2. In type 2 diabetes mellitus, against the background of a decrease in the number of macrophages with the CD163 marker and the concentration of the cytokine TGF-β1, which have an anti-inflammatory effect, in the pancreatic islets, a decrease in the number of β-cells and their functional activity was observed, while the content of α-cells synthesizing glucagon increased. After administration of the sodium salt of 5-amino-2,3-dihydrophthalazine-1,4-dione, the opposite picture was observed in the pancreatic islets: against the background of an increase in the number of CD163+ macrophages and the content of TGF-β1, the number of β cells increased and the number of α cells decreased-cells. The increase in the number of insulin-synthesizing cells was not accompanied by their mitotic activity. It is likely that a decrease in the number of CD163+ macrophages and the level of the antiinflammatory cytokine TGF-β1 in the islets are factors contributing to changes in the cell microenvironment and, as a consequence, the differentiation of β-cells into α-cells. On the contrary, an increase in the number of CD163+ macrophages and TGF-β1 against the background of administration of the sodium salt of 5-amino-2,3-dihydrophthalazine-1,4-dione presumably promotes reverse differentiation of α-cells into β-cells and restoration of insulin synthesis pancreas. Targeted effects on the microenvironment of cells in the pancreatic islet in type 2 diabetes mellitus may be a new approach to treating the disease.

Mental disorders often accompany autoimmune diseases, for example, since 1949 it has been known about “myxedematous madness”, a psychosis caused by hypothyroidism. The most common cause of hypothyroidism is Hashimoto's autoimmune thyroiditis. It is also known about another neuropsychiatric disorder associated with autoimmune thyroiditis, Hashimoto's encephalopathy. It is a severe dysfunction of the central nervous system, the pathogenesis of which is not associated with hormonal disorders. Cytokines are regulators and participants of inflammation, including autoimmune. Certainly, when we are talking about high concentrations cytokines, we mean systemic inflammation. The minimal or mediocre fluctuations in cytokines within the ranges that are characteristic of healthy status or normergic acute phase response in disease cannot be interpreted from the point of view of binary endocrinological logic. In the CNS, cytokines are able to influence on the neuroendocrine control of systemically regulated functions. It is also important that glial cells (astroglia, microglia) are capable of producing a number of cytokines and can affect neurons and develop behavioral changes. In addition, the ability of a number of cytokines outside the CNS itself to act on vagal afferents and through them to convey information to the CNS, affecting its state and functions, has been proven. It is reasonable to assume that minimal fluctuations in cytokine levels may also affect the state and function of the CNS. The aim of the study was to investigate the levels of cytokines in patients with thyroiditis; in patients with thyroiditis associated with mental disorders; in a group of healthy individuals; and evaluate the effect of cytokine levels on clinical manifestations. In the group of patients with thyroiditis and mental disorders, the levels of CCL20/MIP3α, IL-13, IL-2, IL-27, IL-5 were significantly higher than in other groups. At the same time, no positive correlation was found between the clinical manifestations of mental disorders and the levels of cytokines. A positive correlation was found between the levels of some cytokines and free triiodothyronine, as well as the level of antithyroid antibodies. Mental disorders associated with autoimmune thyroiditis may be associated with changes in the cytokine profile and result from neuroinflammation.

The scavenger receptors (SRs)) include > 30 different molecules structurally classified into 11 classes (A to L). They are expressed mostly on stromal macrophages, and their expression may be augmented in direct dependence with concentrations of their ligands. The SRs are heterogenous by their structure, however, being common in their functional potential. E.g., different SR classes may participate in absorption of modified low-density lipoproteins and glycated proteins, apoptotic and ageing cells, altered erythrocytes and platelets, like as a big variety of other endogenous ligands from metabolic and cellular “trash”. A common property of SRs is their participation in removal of small pathogen amounts from blood circulation, regulation of cell and tissue stress responses, ability to form complicated receptor complexes with other receptor types including integrins and toll-like receptors. Opposite to classic pattern-recognizing receptors, the SR involvement does not always elicit a pronounced cellular activation and development of pro-inflammatory cellular stress. The SR functional effects provide interactions between different physiological events and immune system, including the processes of neuroendocrine and metabolic regulation. These mechanisms provide both homeostatic stability and, likewise, act at the border of normal and pathological conditions, i.e., participating in pathogenesis of transitional processes, e.g., physiological ageing. Moreover, the SR-associated processes represent a key pathogenetic factor in different somatic diseases, e.g., those associated with low-intensity chronic inflammation, including obesity, type 2 diabetes, atherosclerosis, arterial hypertension, various neurodegenerative disorders. Similarly, the SRs are involved into the processes of cancer transformation and antitumor response, different processes of classical inflammation, from antigen presentation to the morphofunctional T cell and macrophage polarization in the inflammation foci and immunocompetent organs. SR are playing a controversial role in development of acute systemic inflammation, the main reason for lethal outcomes in the intensive care wards. Targeted effects upon the SRs represent a promising approach when treating a broad variety of diseases, whereas detection of membrane-bound and soluble SR forms could be performed by means of diagnostic and monitoring techniques in many human disorders.

High occurence of liver diseases (toxic, viral hepatitis, liver failure, cirrhosis) requires urgent search of new methods for management of the hepatobiliary diseases. At the present time, the role of immune mechanisms in pathogenesis of diffuse toxic liver damage is not finally clarified. The model of toxic hepatitis induced by carbon tetrachloride (CCl4) is widely known, but this approach allows us to perform complex evaluation and develop the methods for adequate correction of liver disorders in experimental model, which is not always feasible in clinical setting.To design a model of diffuse toxic liver damage, the CCl4 oil solution was used, having been administered intraperitoneally to experimental animals, at a single dose of 50 mg per 100 g body mass. Aiming for correction of toxic liver damage, the injections of aminophthalhydrazide (APH) to experimental animals were carried out intramuscularly at the dose of 2 mg/kg over the terms of experiment. An evaluation of the role of sinusoidal cells (SC) and cytokine production at the local and systemic level were carried out in the model of toxic liver damage caused by CCl4 and its correction by APH treatment. In the course of developing diffuse toxic liver damage induced by CCl4, the production of proinflammatory cytokines TNFα, IL-1α and IL-18 was enhanced at the local level, whereas an increase in TNFα concentration was observed in blood plasma. Following aminophthalhydrazide (APH) administration, the concentrations of proinflammatory cytokines (TNFα and IL-18) decreased at system level, along with locally decreased levels of IL-6 and IFNγ.Changes in the functional state of immunocompetent cells, which include sinusoidal cells (SC), have a significant impact on the development of pathological processes in the liver. The results of our study presume that, over the early periods of toxic impact upon liver tissue, the number of SCs increases both due to influx of blood monocytes and mature macrophages from the peritoneal cavity that enter the injury site directly via mesothelial layer. The SCs provide phagocytosis of damaged hepatocytes and contribute to resolution of the inflammatory process.Modulation of the macrophage activities by APH contributes to increased amounts of SCs at the early stages, and stabilizes their quantities after 2 weeks of APH injections. Change in the numbers of liver SCs during toxic damage affects the production of cytokines. A direct effect of APH upon the SCs may change the production of regulatory factors and compensate the insufficient rate of recovery processes after the toxic damage.

Juvenile idiopathic arthritis is a chronic inflammatory disease of the joints in children, mainly of autoimmune or auto-inflammatory nature. It is a heterogeneous group, which includes different subtypes of the disease. Different mechanisms may play role in the pathogenesis of distinct subtypes of juvenile arthritis. However, a long-term imbalance of pro- and anti-inflammatory cytokines is important for all subtypes of disease. The aim of the present study was to determine spontaneous and stimulated anti-inflammatory cytokines production by peripheral blood cells from the children with juvenile idiopathic arthritis. Patients of 2 to 17 years old with different subtypes of juvenile idiopathic arthritis (n = 99) and healthy children without signs of autoimmune diseases (control, n = 31) were examined. Spontaneous and phytohemagglutinin-stimulated concentrations of IL-1ra, IL-4, IL-10, TGF-β in supernatants of whole-blood cultures were determined by ELISA. Differences in the spontaneous and mitogen-stimulated secretion of the cytokines in patients with different subtypes of juvenile arthritis have not been revealed. The spontaneous IL-1ra, IL-4 and IL-10 production by blood cells in the common group of patients with juvenile idiopathic arthritis was similar to the controls. The median value of spontaneous TGF-β concentration in the patients was below the detection level, whereas blood cells of healthy children had a higher potential of spontaneous TGF-β production. IL-4 and IL-10 production after incubation of peripheral blood cells with phytohemagglutinin in patients and in the control group did not differ from the controls, while IL-1ra and TGF-β synthesis was significantly lower than in healthy children.The spontaneous and/or stimulated production of IL-1ra, TGF-β by blood cells in children with juvenile idiopathic arthritis reflects the pathogenic significance of these cytokines in disease. Stimulation of cells can reveal a latent deficiency in the synthesis of cytokines, which is not evident when determining its concentration in serum or supernatants of spontaneous whole-blood cultures.

A comprehensive comparative evaluation of the ability of L-17 compound (2-morpholino-5- phenyl-1,3,4-thiadiazine) and antioxidant lipoic acid (LA) to correct blood cytokine levels, metabolic disorders and morphological alterations in pancreas in alloxan-induced diabetes was performed for the first time, using an experimental in vivo model. All the tested compounds have been found to correct hyperglycemia and decreased accumulation of glycated blood proteins. The tested L-17 compound belongs to the 1,3,4-thiadiazine series and is able to correct metabolic disorders occurring in alloxan-induced diabetes comparable to activity of LA antioxidant. However, L-17 reduces the cytokine levels to the values of intact animals, that preventing development of systemic inflammatory response which causes damage of target organs in diabetes mellitus.

This review article concerns a challenging problem of HIV infection, immune activation. The processes of immune activation largely determine CD4+T cell depletion, leading to development of AIDS-associated and non-AIDS-related diseases. Immune activation indices are also strong predictors of the clinical outcome. Activation in HIV-infection affects both innate and adaptive immune cells, leads to increased proinflammatory cytokine production and blood coagulation. The reasons for immune activation may include different pathogens (HIV, cytomegalovirus, Epstein–Barr virus, hepatitis C virus), lymphopenia and lymphopenia-induced T lymphocyte homeostatic proliferation, transfer of microbial products from the gut, due to profound CD4+T cell deficiency in lamina propria and altered permeability of intestinal barrier. Other factors that promote the process of immune activation are as follows: T lymphocyte "bystander" activation, increased T cell turnover, and systemic inflammation development. The review covers pathogenic mechanisms HIV-infection associated with immune activation, like as description of different laboratory parameters characterizing its manifestations in HIV-infected patients. Significance and prognostic role of these parameters in assessing efficiency of antiretroviral therapy, development of complications, and adverse outcomes of infection are presented as well.

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.

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. T-lymphocytes play an important role in elimination of tumor cells, in reactions of a transplant against graft and graft versus host disease, in slow-type hypersensitivity, and other reactions directed for maintenance of homeostasis. Along with CD3, an antigen-specific T-cellular receptor (TCR) is another common marker of T-cells. There are two types of TcR – αβ-TcR and γδ-TcR that differ in ontogenetic and functional properties. γδ-T-cells play a significant role in protection of organism against various types of infections, and determination of their amounts should be an integral part of the analysis of patients’ immune status. To these purposes, a multi-colour analysis shuld be used, applying the following combinations of monoclonal antibodies: CD3/CD4/CD8/CD45 and αβ-TcR/γδ-TcR/CD3/CD45. Multi-colour staining and multi-step gating allow of carrying out multiparametric analysis of peripheral blood with high accuracy and reliability. The proposed approach considerably facilitates interpretation of results obtained, and it allows of judging about immune system functioning in various pathological conditions.

Immunomodulating ability of a bacterial preparation (Immunovak VP-4 vaccine), and a compound of eukariotic origin (profetal), the main active component of human α-fetoprotein were under investigation in present study. Culturing of mononuclear leukocytes with the immunomodulatory factors promotes quantitative increase of the cells that express surface antigens specific for cytotoxic lymphocytes, natural killers and natural killer T-cells. Both bio-active preparations possess an expressed anti-cancer effect in vitro, i.e., they induce activation of killing properties of human peripheral blood mononuclear leukocytes, which is accompanied by acquiring their characteristic phenotype.

Abstract. Examination of immunomodulatory actions and clinical efficiency of myelopidum when included into complex therapy that was carried out in 24 male patients with severe (stage 3) penetrating eye injury in the course of trauma treatment. The levels of C-reactive protein, lactoferrin, interleukin (IL)-1β, IL-6, and activity of complement system were measured in peripheral blood, and concentrations of lactoferrin and IL-8 were determined in tears. An increase in lactoferrin, C-reactive protein, IL-1β, IL-8, and C5 complement component levels was detected during early post-traumatic period, as compared with data from the control group. As compared to effects of steroid and non-steroid anti-inflammatory drugs included into standard therapy, treatment with myelopidum has led to a more favorable clinical course of traumatic process, and resulted into more pronounced anti-inflammatory effect that was manifested by decrease in lactoferrin and C-reactive protein levels, reduction of IL-1β concentration, and C5 complement component activity. (Med. Immunol., 2008, vol. 10, N 2-3, pp 239-244).

Abstract. Determination of lymphocyte subpopulations and their phenotypes is an important diagnostic feature, in order to elucidate some disturbances connected with immune system functioning. However, insufficient data are obtained when analyzing only major populations of peripheral lymphocytes. In order to perform clinical diagnostics, the data about minor lymphocytic populations and activated cellular pools seem to be more pertinent.Studies of peripheral blood cell subpopulations of healthy donors performed in different Russian regions allowed to assess quantitative distribution intervals for both major and minor immune cell subpopulations in humans. The results obtained, as compared with data from literature, provide an evidence for similar reference intervals for main immune cell subpopulations in healthy donors, independent on their habitation area.Present work has resulted into development of algorithms for cytometric studies and generation of certain panels of monoclonal antibodies enabling evaluation of all main lymphocyte subpopulations, as well as their minor subsets participating in emerging immune response. The distribution intervals have been estimated for such minor subpopulations, as B1- and B2-lymphocytes, memory B-cells, γδ- and αβT-cells, regulatory and naїve T-cells, cytotoxic and secretory NK-cell polupations.The results of present study, while been performed with peripheral blood of healthy donors, may provide a basis of reference values when studying subpopulation profile of immune cells.

Abstract. The lecture presents modern data on systemic inflammation, as well as an original concept of its genesis. This disorder is considered a typical pathological process being fundamental to pathogenesis of the most critical states of different origin. Distinct features of “classical” and systemic inflammation are discussed, whereas the underlying immunological and pathophysiological mechanisms of inflammation are subject to classification. Main constitutive processes, phases of systemic inflammation, and certain variants of their proceeding are characterized as well. A definition of “systemic alteration” has been specified.

Abstract. Fourteen groups of patients have been investigated and divided into 2 classes. The first class included the following cohorts of patients: relatively healthy persons, age 18 to 55 yrs (n = 50); elderly persons 60 yrs old, as well as senior persons (n = 22); persons with chronic adnexitis, women in their 1st trimester of pregnancy (n = 16); climacteric syndrome (n = 16); autoimmune thyroiditis (n = 29). The second class of patients included following cohorts: elderly persons with chronic cardiac insufficiency (CCI) II-III stage (n=49); valvular cardiac disease (rheumatism, n = 15); psoriatic arthritis (n = 12); reactive arthritis (n = 17); antiphospholipid syndrome, a sub-group in the 1st trimester of pregnancy (n = 5); systemic lupus erythematosus (n=49); decompensated atherosclerosis of femoral artery (n = 38); end-stage renal disease (n = 42). Plasma cytokines (TNFαα, IL-6, IL-8, IL-10), acute-phase C-reactive protein (CRP), cortisol, troponin I, myoglobin, D-dimers, interleukin-2 soluble receptor (IL-2sR), and eosinophil cationic protein (ECP) were determined in all the patients, by means of immune chemiluminescent technique (Immulite; Siemens Medical Solutions Diagnostics, USA). The integral indices of systemic inflammatory reaction (SIR) have been calculated, i.e., a Reactivity Coefficient (RC) and a Reactivity Level (RL). In the patients belonging to Class 1 cohorts, an absence of chronic systemic inflammation features was revealed, despite of some signs of systemic inflammatory response. Meanwhile, a majority of Class 2 patients have shown the signs of chronic systemic inflammation stage I to III.

We have performed immunophenotyping of lymphocytes in 150 HIV-positive patients, by means of flow cytometry. All the persons under study were classified into three groups, depending on absolute contents of CD3+CD4+T-lymphocytes in peripheral blood, according to HIV infection staging in adults and adolescents, as amended by CDC (USA). It was revealed that, at each subsequent stage of disease, a decrease in absolute numbers of T-helpers is accompanied by drop in relative and absolute numbers of lymphocytes, as well as in total T-lymphocyte numbers, and decreased amounts of absolute cytotoxic T-lymphocytes levels, combined with increase in their relative contents, along with decreased CD3+CD4+/CD3+CD8+ ratio, diminished levels of double-positive subpopulation, decrease in relative and absolute levels of αβТ-lymphocytes and significant drop in absolute levels of γδТ-cells, while their relative amounts remained normal. These findings demonstrate a pattern typical to the absence of immunological reactions in response to antigens persisting in the organism.

Abstract. An intracellular staining technique of marker cytokines was applied for evaluation of quantitative distribution of T-lymphocyte effector subsets in children and adults. The results of this study have shown that the numbers of Th2 lymphocytes in infants (7 to 12 months old) correspond to appropriate normal values in adults, being accompanied by decreased Th1 and Тс1cell contents, whereas absolute amounts of Tc2 lymphocytes proved to be higher than in all other age groups. Absolute counts of Th17 and Tnc17 subsets in infants of 7 to 12 months were higher than in adults. Hence, these infants exhibited a shift of immune response towards Th2 (Tc2) and Th17 (Tnc17), thus presenting a potential factor of Th2-mediated allergic disorders in the children under one year. In children over 1 year, absolute amounts of Th1 and Тс1, as well as Th2 and Тс2 lymphocyte counts did not significantly differ from adult values. Th2 lymphocyte counts among adolescents (15 to 18 years) were decreased, along with relative increase of IL-17А-positive lymphocyte scores, thus reflecting a probable predisposition for autoimmune disorders induced at this age. The age-dependent increase in Th1 and Тс1 counts in children may immediately correlate with formation of T-cell memory pool.

Abstract. The lecture presents current data, as well as authors’ view to the issue of immune system involvement into inflammation. General physiological principles of immune system functioning are considered in details. Immunological mechanisms of local inflammation and participation of immune system components are analyzed with regard of protective/adaptive reactions in inflammatory foci. Original formulations of basic concepts are presented from the viewpoint of pathophysiology, immunopathology and clinical immunology, as being applied to the issues discussed. (Med. Immunol., 2011, vol. 13, N 6, pp 557-568)

Abstract. We performed a study of efficiency for various immunocorrective drugs in a group of 548 young schoolchildren with recurrent respiratory infections. Frequency of respiratory infections and complication rates were taken as endpoints in this study. It was revealed, that preventive immunocorrection by bacterial lysates or glucosamine muramildipeptide combined with vitamin-mineral complexes may reduce frequency of respiratory infections and their complications at statistically significant levels, as well as to restore some abnormal parameters of immune profile, i.e., CD3, CD4, CD16, induced NBT test, IFNγ, TNFα and IgG. Preventive use of Echinaceae purpurae herbae succus or interferon alpha-2b in combination with vitaminmineral complexes statistically significantly reduces only the frequency of respiratory infections, and partially restores some deficient parameters of immune status (IgG, TNFα, CD16). Introduction of preventive immunocorrection in childhood institutions, with > 90 % coverage of children with recurrent respiratory diseases is associated with a decrease in frequency of respiratory infections not only in this cohort, like as among general population of the same age group.

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. The role of hepatitis C virus (HCV) co-infection in violation of restoring CD4+T cells in HIVinfected patients receiving antiretroviral therapy (ART) was investigated. Seventy eight patients with effective virological response to ART (viral load < 50 copies/ml) who either had or had no good immunological CD4+Tcell response after two years of treatment were recruited. Twenty one relatively healthy volunteers served as controls. The numbers of main T-lymphocyte populations and their apoptosis rates were determined. Thymus productive function was assessed by the number of αTREC-positive CD4+ and CD8+T cells. It is shown that HIV/HCV co-infection is characterized by lower numbers of CD4+T-lymphocytes in patients’ blood compared with HIV-monoinfection. This is not associated with the cells’ increased death rates. We also demonstrate that HCV coinfection reduces the production of CD4+, but not CD8+T-lymphocytes, by the thymus.