Abstract. γδТ lymphocytes represent a poorly investigated heterogeneous population of T cells that are found, predominantly, in mucosal structures, and express common characteristics of innate and acquired immunity. Lack of both antigen processing and MHC-restriction determines an ability of γδТ cells to identify broad spectrum of antigens, the origin of which is yet unknown, and the recognition mechanism have been not established yet. A variety of biological functions, first of all, cytolysis, immune regulation, antigen presentation and repair of tissue damage, define a unique role of this population in infectious diseases, tumors, and autoimmune disorders. Nowadays, principal efforts of scientists are directed to investigation of γδТ cell therapeutic potential, search and production of γδТ cell agonists, as well as design and optimization of herapeutic protocols that may be targeted to γδТ lymphocytes.

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.

Abstract. An alternative pathway of L-tryptophan biotransformation is provided by the indolamine 2,3-dioxygenase (INDO), and it results into synthesis of kynurenine and other «distal» metabolites, playing a pivotal role in immunoregulation and down-regulation of immune inflammation. PBMCs from healthy volunteers, patients with progressive vulgar psoriasis (PASI (M±SD) = 25.6±16.6), or patients with psoriatic arthropathy (PASI = 40.3±24.5). The cells were incubated for 24 hrs with IFNγ (500 IU/ml, stimulated cultures) or without cytokine (control cultures). Distinct abnormalities in spontaneous and induced kynurenine production (colorimetric method) and INDO expression (semi-quantitative RT-PCR) were observed in psoriasis and psoriatic arthritis. PBMCs from psoriatic patients, in comparison with healthy donors, were characterized with slightly increased spontaneous kynurenine production, spontaneous and IFNγ-induced INDO expression, while IFNγ-induced kynurenine levels were approximately two times higher. In psoriatic arthritis, spontaneous kynurenine production and INDO expression were significantly lower than in donor’s PBMC, whereas IFNγ-induced kynurenine production were the same as for the donors, while IFNγ – induced INDO expression was markedly increased.

Abstract. The article deals with results of studying combined effects of ethambutol and protein antigen of Мycobacterium tuberculosis upon proliferative response and cytokine-producing ability of peripheral blood mononuclear cells in patients with infiltrative lung tuberculosis prior to beginning of specific antibacterial therapy. The mononuclear blood cells were cultivated with complex protein antigen of Мycobacterium tuberculosis and ethambutol. In the patients with lung tuberculosis, a decrease in proliferative reserve of lymphocytes was detected, in absence of increased IL-2 and IL-10 secretion. The combined effect of ethambutol and protein antigen of Мycobacterium tuberculosis upon peripheral blood mononuclear cells in the patients with pulmonary tuberculosis and healthy donors was similar, and it was expressed as antiproliferative effect and selective suppression of IFNγ production.

Abstract. The authors suggest a hypothesis about possible role of specific immune reactions in pathogenesis of severe clinical forms of dysplastic coxarthrosis. It is proposed to apply relative and absolute lymphocytosis scores in peripheral blood, as well as increased levels of serum IgA, as screening indices that allow of classifying the patients into the high-risk group for possible post-surgical complications.

Abstract. A study of leukocyte regulation system in a remission stage of bronchial asthma (BA) patients included following tests: CD-typing of lymphocytes, investigations in humoral and phagocytic mechanisms of immunity, evaluation of bronchoalveolar lavage characteristics, measurements of spontaneous production of IL-4, IL-1β, IL-10 and IFNγ cytokines in whole blood supernates and bronchoalveolar lavage, as compared with control samples. The first group represented mild-stage, mixed-type BA patients (both allergy- and infection-dependent), being in remission state. The second (control) group consisted of sixteen conditionally healthy patients.

The results have shown that, even at early stages of BA development, some components of chronic inflammatory process are formed. They could be characterized as accumulation of different inflammatory cells, i.e., eosinophils, neutrophils, alveolar macrophages and monocytes at the local level. In parallel, some changes in T-, B-subpopulations and phagocytic compartment occur at systemic level. All these events comprise a basis for changes in type and quantity of the cytokines produced. It was established that the profile of cytokine secretion in peripheral blood is identical to the cytokine secretion profile in lung tissue.

Abstract. The aim of the study is to specify the levels of STAT6, and phospho-STAT6 under the influence of IL-4 in patients with bronchial asthma (BA). The samples from ten healthy controls and thirty-three BA patients with allergic and non-allergic clinical forms of different severity were under investigation. Peripheral blood lymphocytes were treated with 10 ng/ml of IL-4 (Sigma Aldrich, USA) for 1 h. Then the proteins (STAT6 and phospho-STAT6) expressed in peripheral lymphocytes were analyzed by Western blot of cell lysates. Preparation of the cell lysates and Western blotting were carried out using standard procedures. Antibodies against phospho-STAT6 and STAT6 (10 ng/ml) were used (Cell Signaling, USA). Levels of thespecific proteins were standardized according to β-actin (Cell Signaling, USA). Treatment with IL-4 caused an increase of phospho-STAT6 levels in lymphocytes of all BA patients, as compared with control group. In allergic BA, the phospho-STAT6 levels were significantly higher than in non-allergic clinical forms. Expression of STAT6 in lymphocytes of patients with severe BA was significantly higher, as compared to BA of moderate severity. An IL-4-induced activation of the STAT6 transcription factor was revealed in an in vitro system, being mostly expressed in allergic BA. The level of STAT6 may serve as a BA severity index. This study was supported by a «Scholarship of the Year» grant from the St. Petersburg State Medical I. Pavlov University (2007).

Abstract. We studies blood sera of the women with verified endometriosis (End), cancer of ovaries (Can), cystadenoma (Cyst) and healthy women by similar age (Cont) for the contents of cytokines (TNFα, IL - 1β, IL-6, IFNγ), some macroglobulin proteins transporting them across cell walls, i.e., α2-macroglobulin (α2 - Mg), and pregnancy-associated α2-glycoprotein (α2-PAG)), as well as some functional forms of α2 - Mg (α2-Mg-IgG and α2-Mg-plasmin complexes (α2-Mg-Pl), and intrinsic components of these complexes (total serum IgG and plasmin)), to establish their role in pathogenesis of abovementioned diseases. We have shown that the direction and expression of the changes are quite different. I.e., in ovarian cancer, the levels of α2 - PAG, α2-Mg-Pl, IL-6, and TNFα were considerably increased, concentrations of α2-Mg-IgG complex are moderately increased, but the contents of total IgG and IFNγ are not changed. Meanwhile, the levels of total α2-Mg and Pl are reduced. In cystadenoma, the total α2-Mg levels are significantly reduced, but total Pl and α2-PAG concentrations are constant, and only IL-1β and both type of complexes are significantly increased. Endometriosis is characterized by considerable increase of α2-PAG, IL-6, IL-1β and IgG serum levels, accompanied by relatively high concentrations of α2-Mg-IgG and α2-Mg-Pl complexes. To our mind, the changes revealed may represent either consequence, or a factor of an active neoplasm progression.

Abstract. Phenotypic and functional features of IFNα-induced dendritic cells (DCs) were studied in patients with chronic viral hepatitis B and C (HBV and HCV), and in cases with hepatitis-related liver cirrhosis (LC). It was shown that DCs are characterized by delayed differentiation/maturation which was more pronounced in HCV patients, as well as in all patients with LC, regardless of virus type. DCs from HBV patients were characterized by increased IFNγ secretion. Transformation of HBV-infection to LC is accompanied by a moderate decrease in IFNγ production, combined with a significantly increased IL-10 secretion. Irrespectively of fibrosis severity, the IFNα-induced DCs of HCV patients displayed active IL-10 synthesis. Moreover, ability of DCs to secrete IFNγ was significantly decreased only in cases of fibrosis-complicated HCV-infection. With respect to TNFα and IL-4 production levels, DCs of the patients were compatibe to normal donor cells, independently on the type of virus, or fibrosis severity. DCs from HBV- and HCV-patients were characterized by intact allostimulatory and Th1/Th2-stimulatory activities in MLC. At the same time, IFNα-induced DCs exhibited suppression of allostimulatory and increase in Th2-polarizing activity upon LC development, both in HBV and HCV patients.

Abstract. We present some results concerning investigation of humoral immune factors (virus-specific antibodies, complement components, total serum immunoglobulins, circulating immune complexes), and the levels of tick-borne encephalitis (TBE) viral antigenemia in the persons subjected to stings of ticks. Elimination of the virus in vaccinated people with short-term antigenemia, with higher indices of innate humoral immunity and virus-specific antibodies, proved to be faster while being more complete than in non-vaccinated persons. In cases of a long-term TBE antigenemia, both vaccinated and non-vaccinated people showed low levels of virusspecific antibodies along with decreased natural resistance, thus, probably, contributing to delayed elimination of TBE virus.

Abstract. Interferon profile was studied in 412 children and 41 adults with acute respiratory virus infections, as well as in forty-eight healthy children and ninety-nine adults. Appropriate interrelations were evaluated between interferon contents, and age, specific clinical features, etiology and clinical course of the disease. Therapeutic efficiency of interferons and interferon inducers was studied at 928 children. Some novel parameters are proposed to assess interferon state, i.e., stimulation index of interferon production, and interferon-producing activity of lymphocytes that represent individual functional activity of the patients’ immunocompetent cells. Criteria for interferon administration.

Abstract. Polyclonal immunoglobulins G (subclasses 1, 2, 4) from sera of 255 patients and 69 healthy persons were studied by a combined approach using rivanol treatment and affinity chromatography. Enzymatic reactions were carried out according to the methods that we have previously developed and validated for evaluation of abzyme activity in the patients with different disorders. The levels of DNase, proteolytic BAPNA-amidase (benzoylarginine-p-nitroanilide amidase), and superoxyde dismutase abzyme activity in spondyloarthropathies proved to be significantly higher (p = 0.001), as compared with a control group. Catalase activity of immunoglobulines in the disorders studied was compatible to control levels (p > 0.05). Analysis of relations between abzyme activity and clinical and laboratory signs of the diseases has revealed some significant correlations. Prevalence of abzyme DNAse activity is found in the patients with psoriatic arthritis, as compared to reactive arthritis and ankylosing spondilitis (p < 0.001).

Abstract. Treatment with statins results in reduction of disease activity in one-third of patients with rheumatoid arthritis (RA). The aim of this study was to assess some factors that may predict clinical response to simvastatin therapy before starting the treatment. We evaluated an association of treatment efficacy with baseline clinical and laboratory parameters including disease activity measures, cytokine profiles in sera and culture supernatants of peripheral blood mononuclear cells. Thirty-three patients with active RA were enrolled in the study. The patients were treated with simvastatin at 40 mg/day for three months. Eleven patients (33%) developed a moderate response according to EULAR criteria. It was shown that serum IL-10 concentrations was higher in responders, and positively correlated with clinical response to simvastatin. We carried out a receiver operating characteristic curve (ROC) analysis in order to assess the accuracy of serum IL-10 for the predicting of EULAR response development. The cut-off threshold corresponding to the highest sensitivity (89%) and specificity (62%) was a value of 6.5 pg/ml. In conclusion, the performance characteristics of serum IL-10 measurement proved to be good enough to predict EULAR response to simvastatin therapy in RA patients.

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.

Absract. A research of immune profile was carried out in patients with chronic ischemic disease of gastrointestinal tract. The studies included evaluation of subpopulational profile of lymphocytes, estimation of functional activity of neutrophils, measurement of immunoglobulin amounts, and cytokine levels in peripheral blood. Immune status of the patients suffering from chronic ischemic disease of digestive tract, depends on the level of gastrointestinal tract damage. A predictive self-descriptiveness of a number of immune parameters has been revealed among patients.

Abstract. The phenotype and functional properties of antigen-presenting cells (APCs: blood monocytes and in vitro generated macrophages/dendritic cells) were investigated in patients with pulmonary tuberculosis (TB, n = 192) with different levels of proliferative response to M. tuberculosis antigens (PPD-responsive vs PPD - anergic patients, n = 118 and 74, respectively). A functional deficiency of all 3 types of APCs was revealed in patients with TB. I.e., a monocyte disfunction was displayed by low CD86 and HLA-DR expression, 2-fold increase of CD14+CD16+ subset, high level of FasL+ and IL-10+ cells, and enhanced IL-10 and IL-6 production upon LPS-stimulation. The in vitro generated macrophages from blood monocytes challenged with GM-CSF, were characterized by shifted Th1/Th2 balance (down-regulated production of IFNγ and IL-18 combined with up-regulation of IL-6 and IL-10), and reduced allostimulatory activity in mixed lymphocyte culture. The dendritic cells were characterized by decrease of mature, activated CD25+ cells, low level of IFNγ production in conjunction with enhanced capacity to produce IL - 10 and IL-6, and profound reduction of functional (allostimulatory) activity. The APC disfunction of were most prominent in PPD-anergic patients. A possible role of APC disfunctions in disturbed antigen-specific T-cell response to M. tuberculosis is discussed.

Abstract. Anti-ergotypic response is considered to contribute significantly to the regulation of immune response. Ergotope-associated antigenic determinants include molecules upregulated on the surface of activated T cells (CD25, hsp60 and others). Anti-ergotypic cells are directed against these surface determinants usually complexed to MHC-I and/or MHC-II. Here we demonstrate regulated expression of HLA-Dr, СD25, hsp60 and hTERT mRNA during in vitro activation of peripheral T-lymphocytes with anti-CD3 antibodies and IL - 2, with reactivation on day 7. The percentage of CD25-expressing T cells showed sharp increase as early, as by the day 3 of activation, and it varied only slightly at later terms. The hsp60 levels rose as well, reached a peak at day 3, and gradually decreased thereafter. HLA-Dr expression was induced upon the activation, with a peak at the days 8 or 10. hTERT mRNA amounts also increased, as compared with baseline values, showing two peaks at the days 3 and 8. Anti-ergotopic response may control autoimmunity by targeting the activated T cells, regardless of their specificity. Therefore, immunotherapy of different autoimmune disorders aiming to activate anti-ergotopic responses, may be of a sufficient clinical interest.

Abstract. Ability of cannabinoid receptor ligands WIN 55.212-2 and anandamid to inhibit synthesis of TNFα and IL-8 was studied in healthy donors and men with allergic disorders. To establish mechanism of action for investigated substances, the selective antagonists of the СВ1-receptor (SR141716A) and for СВ2 - receptor (SR144528) were applied. Studies with whole blood dilutions allowed of approximating in vivo conditions when investigating biological properties of WIN-55.212-2 and anandamid. The synthetic cannabinoids WIN - 55.212-2 and anandamid at a concentration of 3-10 μМ were capable of reducing synthesis of TNFα and IL-8 in lipopolysaccharide-stimulated blood leukocytes, both from healthy donors and subjects with allergic disorders. It was revealed that the antagonist of СВ1-receptor (SR141716A) did not exert a receptor-mediated effect for WIN-55.212-2 and anandamid. Meanwhile, a СВ2-receptor antagonist (SR144528) entirely eliminated completely the blocking effect of anandamid and WIN-55.212-2.

Abstract. In the present study, blood levels of circulating immune complexes and of their pathogenic subpopulations were determined in rats following ionizing irradiation. Experimental animals were treated with synthetic Schiff base aromatic amino acid derivatives, nicotinyl-L-tyrosinate or nicotinyl-L-tryptophanate, before irradiation, whereas untreated irradiated rats served as controls. The results obtained demonstrate significantly increased levels of immune complexes, as well as presence of a pathogenic subpopulation of circulating immune complexes in blood of irradiated animals. In addition, the data obtained suggest a normalizing effect of nicotinyl-L-tyrosinate and nicotinyl-L-tryptophanate upon the mentioned parameters. On the basis of these observations, a cyto- and immunoprotective ability of nicotinyl-L-tyrosinate and nicotinyl-L-tryptophanate may be proposed.

Abstract. The study performed has shown a positive clinical and immunological dynamics in forty children with medium-stage atopic bronchial asthma in remission stage, after a course of adaptation for periodical hypobaric hypoxic stimulation. These changes were expressed as a reduction in IgE, circulating immune complexes, Interleukin-4, and rise in Interferon-γ and Interleukin-18 contents. A retrospective analysis has shown that a more stable positive clinical and immunologic effect is observed in the group of patients with relatively low IgE levels observed after hypobaric hypoxic stimulation.

Abstract. The subpopulation structure of immunocompetent cells of peripheral blood, concentration of IgA, IgM, IgG, IgE and serum levels of IL-4 and IL-10 for the patients was studied. Reduced contents of CD8+, CD56+ lymphocytes and increased contents of CD4+, CD22+ lymphocytes, serum levels of IgA, IgM, IgG, IgE are revealed in Hkakass with asthma compared to healthy Hkakass. It was also revealed in Hkakass with asthma that contents of CD3+ and CD8+ lymphocytes, serum concentration of IgA, IgE increased compared to Caucasoids with asthma. The serum level of cytokines IL-4 were found to be considerably increased in Hkakass with asthma compared to healthy Hkakass and to Caucasoids with asthma. On the other hand, the comparison of concentration of IL-10 showed that serum level was higher in Caucasoids with asthma than Hkakass with asthma.

Abstract. We have performed immunophenotyping of lymphocytes, employing a flow cytometry approach, along with evaluation of spontaneous or mitogen-induced production of cytokines (IFNγ, IL-4, IL-1β, IL - 10), in the patients with atopic and bacterial (mixed-type) bronchial asthma. We have detected mostly similar changes in lymphocyte parameters. However, among the patients with bacterial asthma, an imbalance of immune cells was more pronounced. Some common alterations of cytokine production were found, i.e., a decrease in IFNγ and IL-1β production, along with increased IL-4 and IL-10. These changes were more significant in the subjects being in acute phase of either atopic, or bacterial asthma. Moreover, IL-10 suppressed IFNγ to higher degree, than IL-4, thus favoring distinct predomination of Th2-type immune response.

Absract. The data are presented that concern the levels of IL-1 receptor antagonist (IL-1 Ra) in blood serum of twenty-five patients with severe surgical sepsis. Sixteen patients recovered from the condition, whereas lethal outcome was registered in nine cases. The study was performed by means of immunoassay, using «IFA-Best IL-1 Ra» kits supplied by the Vector-Best firm. Only three patients showed IL-1 Ra contents within normal limits, whereas others exhibited marked increase in IL-1 Ra levels, up to a mean of 2443.656±382.85 pg/ml (normal range, 50 to 1000 pg/ml).

Abstract. Sixteen children from 3 to 6 years old with complicated premorbid background, carrying S. pneumoniae, who had inflammation markers (altered IL-6 and TNFα amounts in peripheral blood analysis)тalong with immunological deficiency, were vaccinated with a «Pneumo 23» preparation. A good tolerance of «Pneumo 23» vaccine was determined. A significant increase of specific antibodies was established both to pneumococcal polysaccharides and to polysaccharides, containing in «Pneumo 23». A tendency to diminished inflammatory events and activated immunological response was revealed, that was expressed as a decrease of serum IL-6 and increase in TNFα.