It is obvious that any evolving scientific medical field is a dynamic system that cannot stay at the stage of accumulation of primary information, and inevitably goes to the stages of clinical trials, generalization of information in meta-analyses and completes the study by creation of practical guidelines. The purpose of this study was a quantitative analysis of publicly available data in the field of clinical immunology in Russia during 2008-2015, identifying the ratios of clinical trials, meta-analyses, and practical guidelines, as well as evaluating the results by comparison with other BRICS countries. Study design was performed by retrospective bibliometric methods. It is revealed, that, in Russia, 16 clinical trials, 3 meta-analyses and 1 practice guideline were issued per 1000 original journal articles. Accordingly in the People’s Republic of China this ratios have made 34/25/4; in Federal Republic of Brazil, 42/87/7; in Republic of India, 76/58/34, and in Republic of Southern Africa, 134/43/36. Moreover, we have obtained evidence which suggests optimistic prospectives for scientific clinical immunology in Russia.

 

Immune system plays a key role in pathogenesis of ischemic stroke. Ischemic brain injury causes destruction of cells and extracellular matrix proteins, and the release of „danger“ molecules that cause rapid activation of innate immunity, and induce adaptive autoimmune reactions. The cells of innate immunity are responsible for elimination of cellular and extracellular debris and tissue repair, being, however, involved in post-ischemic inflammation causing brain injury. An adaptive immune response which develops within a few days is directed against self-antigens, being not related to nervous tissue damage in acute phase of stroke. The cells of adaptive immunity are able to regulate microglial activity and promote repair processes in the central nervous system. However, long-term persistence of lymphocytes with reactivity for brain-derived antigens may be involved into the atrophy of cerebral cortex, and development of dementia in remote post-stroke period. Immune response following ischemic brain stroke is manifesting by local and systemic reactions. Nevertheless, in contrast to prolonged local intracerebral inflammation in infarction area, and peri-infarction region, a systemic inflammatory response is quickly replaced by development of immunosuppression, as evidenced by lymphopenia, lymphoid organ atrophy and defective functions of immune cells. Immunosuppression is largely due to activation of sympathoadrenal system, aiming for limitation of inflammatory and autoimmune reactions. However, excessive immunosuppression leads to development of infectious complications, which may often cause lethal outcomes, and also may have a negative impact on efficiency of repair processes. The present review provides experimental and clinical data describing the innate and adaptive immune reactions in response to cerebral ischemia. Moreover, this review discusses possible pathogenetic importance of immune cells and their positive or negative effects, thus allowing further development of new therapeutic approaches for treatment of ischemic stroke.

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.

 

Effect of γс-cytokines (IL-2, IL-7 и IL-15) upon maturation and differentiation of cytotoxic and helper CD45RО⁺ Tcell population was studied in the homeostatic in vitro culture conditions. We have found that IL-2, IL-7 and IL-15 mediate maturation and differentiation of CD8 T cells central memory, replenishing the population of cells that exhibit effector functions. Action of IL-2 on CD4⁺ central memory T cells is accociated with generation of effector memory cells by reducing the number of immature effectors (CD62L–CD27⁺), whereas IL-7 promotes the formation of immature (CD62L–CD27⁺) effectors. IL-2, IL-7 and IL-15 initiate formation of terminally-differentiated cells in a subpopulation of CD8⁺CD45RO⁺ lymphocytes, providing a stable immunological memory to a pathogen reinfestation.

 

Studies in serum concentrations of cytokines was performed in 103 high-ranked athletes from the sports different by energy consumption (bobsleigh and shooting sports). We have shown that the cytokine concentrations (IL-4 и IL-18) in bobsleigh sportsmen were sufficiently higher than in shooters. I.e., the IL-4 concentration was 1.5±0.9 pg/mL in bobsledders, and 0.45±0.23 pg/mL in shooters (р < 0.05). The IL-18 concentration was 467.5±155.2 pg/mL in bobsledders and 304.5±126.8 pg/mL in shooters. Meanwhile, the IL-6 and IL-10 in blood serum showed only a tendency for increase. The IFNγ concentration in bobsledders did not differ from similar parameters in shooters. When comparing the data in females, the IL-4 and IL-10 figures were 3.7-fold higher in bobsledders for IL-4, and 2.34-fold higher for IL-10, when compared to the shooters. Analysis of cytokines in blood of athletes with high energy consumption has shown significant fluctuations of the given parameters in athletes of both sexes. We have not revealed any correlations between the cytokine contents in blood serum and main anthropometric parameters (body muscle mass, body index mass, energy comsumption). Moreover, increased contents of the cytokines was found in bobsledders at more intensive physical loads. Hence, the energy consumption influences the cytokine state parameters. However, all the cytokine values in athletes are within population norms which may due to adaptation of the athletes for high loads which may be determined by, e.g., adequate usage of specialized sport food for their nutrition.

 

Lymphotoxin-α (LTA) is a major pro-inflammatory cytokine produced at the early stages of vascular inflammation, taking part in the formation of arterial atherosclerosis and development of coronary heart disease. Functional changes in the gene encoding LTA production may influence the development of coronary heart disease with unfavorable progression. However, studies for associations between rs1041981 (C-804A) LTA gene variant and development of acute cerebrovascular accidents, myocardial infarction, and severity of coronary atherosclerosis have yielded contradictory results. The purpose of our study was to investigate an association of rs1041981 gene LTA with risk of adverse events within five years of follow-up in the patients with acute coronary syndrome without ST elevation ST (nonST-ACS). 178 patients with nonST-ACS from the Kemerovo Cardiology Center Registry were included into the study. Genotyping of rs1041981 site variable LTA gene was performed by TaqMan technique using an “iCycler iQ” device (BIO-RAD, USA). Results: we have found that the A allele and A/A genotype polymorphism in LTA gene (rs1041981) have been associated with development of adverse cardiovascular events over five years of observation (respective p levels were 0.02 and 0.036). In patients with A/A genotype, the rs1041981 polymorphism in LTA gene was associated with 3.8-fold increase in adverse cardiovascular events, compared to patients having A/C or C/C genotype. Carriage of A allele in LTA gene (rs1041981) doubles the risk of adverse cardiovascular events in patients with nonST-ACS at long observation terms. By means of Kaplan-Meier method, we have determined that survival to the first endpoint occurred more often in carriers of the genotype A/A of LTA gene (rs1041981). The A/A genotype of LTA gene (rs1041981) proved to be more significant (p = 0.016) for development of adverse outcomes, when combining the patients with A/C and C/C genotypes. One may draw a conclusion that A allele and A/A genotype of rs1041981 LTA polymorphism is associated with development of adverse cardiovascular events during the five-year period following the index event in patients with nonST-ACS.

 

As a preparatory stage for implementation of genetic testing for severe combined immunodeficiency under a neonatal screening program, a study was performed in Sverdlovsk Region which concerned quantitative determination of T and B cell neogenesis markers (TREC and KREC, respectively) in blood of conditionally healthy newborns. Archived samples of dry blood spots collected in test-forms for routine neonatal screening were used as biological material for the study of full-term 26 girls and 26 boys who did not exhibit serious illnesses during first year of their life. In addition, we investigated potential effects of foetal gestational age upon the number of TREC and KREC in preterm infants. Blood samples from 55 preterm infants (23 to 36 gestational weeks) were also examined. It was shown that the levels of TREC and KREC increased sequentially with the increased gestation terms, but the quantitative changes of markers showed different dynamics. In this respect, the recommended terms of blood sample collection for SCID screening is entirely consistent with timing of blood sampling for routine newborn screening. An alternative result was obtained with a complete absence of TREC or KREC in blood sample of a newborn, irrespectively of prematurity degree (at valid copy numbers of a control gene) which should serve as an indication for immediate consulting of the child by immunologist and in-depth immunological examination, because it may be a first prognostic sign of a fatal disease. In order to obtain correct cut-off levels for TREC/KREC, additional studies are needed on a larger sample of newborns (1.000 to 5.000), followed by validation of the obtained reference boundaries in studies involving patients with different forms of primary immunodeficiencies.

 

The goal of present study was to determine the levels of mast cell tryptase (MCT) in whole saliva, and blood serum IgE antibodies in patients with intolerance for dental prosthetic materials (IDM) before and after removal of prosthetic constructs. We have conducted examination of the patients suffering from the IDM symptoms, who were divided into 2 groups depending on the time span between the end of prosthetic treatment and emergence of pathological symptoms: group 1 (n = 19), 1 to 14 days (symptoms emerged immediately after treatment); group 2, (n = 18), IDM symptoms occuring 6 months to 5 years later; group 3 (n = 16), controls without complaints for IDM. Whole saliva (WS) samples were collected before removal of prosthetic constructs and 1 month later. In group 1, salivary MCT was detected in 16 subjects (84.2%) before removal of prosthetic constructs, while 1 month after MCT not detectable in saliva (p < 0.001). Salivary MCT in control group was not detected both before and after removal of prosthetic constructions. Hence, mast cell tryptase in whole saliva could be a diagnostic marker for intolerance to dental materials. In group 1 of the patients, we detected IgE antibodies to Ni-HSA in 78.9% of patients, IgE antibodies to Cr-HSA in 68.4% of patients and IgE to Co-HSA in 52.6% of patients. Salivary MCT levels strongly correlated with IgE levels to Ni-HSA (R_spearman = 0.9; p < 0.05), showing moderate correlation with Cr-HSA (R_spearman = 0.7; p < 0.05). The data obtained suggest some prevalence of immediate type immune reaction against dental materials. Notable local increase of MCT level could be an important diagnostic marker of local inflammatory process initiation. MCT in whole saliva was found only in 3 patients (16.7%) from group 2; those had IgE antibodies to metal ions in blood serum, thus indicating IgE-dependent reaction type. Other patients from group 2 were likely to develop a different type of allergic reaction, e. g. delayed or granulocyte-mediated response. To perform reliable diagnostics of allergy to the components of dental materials, it is reasonable to measure salivary MCT before and after removal of prosthetic constructs. Salivary MCT level monitoring allows to confirm a role of prosthetic constructs in IDM emergence, and a need for their replacement.

 

Resistance of pathogenic microorganisms to conventional antibiotics is growing rapidly in recent years, accompanying with an increase of mortality caused by hospital-acquired infections. Therefore, a search for novel drugs to combat antibiotic resistant bacteria is one of the priorities in biomedicine. Natural compounds which are contained in host defense effector cells of humans and animals, may serve as prototypes for developing principally new antibiotics. Such compounds include antimicrobial peptides of innate immunity, in particular, a group of proline-rich peptides. The aim of this work was to evaluate antimicrobial activity of proline-rich peptides, belonging to bactenecins family, against several clinical isolates of drug-resistant Gram-negative bacteria. The bactenecins under examination (ChBac3.4, ChBac5, mini-ChBac7.5Nα, miniChBac7.5Nβ) have been previously found in leukocytes of a domestic goat Capra hircus. We have shown that chemically synthesized analogs of these peptides exhibited a pronounced in vitro antimicrobial activity against Escherichia coli and Acinetobacter baumannii (minimum inhibitory concentrations (MIC) as estimated by a broth microdilution assay varied between 1 to 4 µM) and, to a lesser degree, against Pseudomonas aeruginosa and Klebsiella pneumoniae (MIC 2-16 µM). It was revealed that antibacterial activity of these peptides may be increased if applied in combination with some conventional, antibiotics. E.g., synergistic antimicrobial effects against E. coli have been shown for mini-ChBac7.5Nα bactenecin combined with amikacin (minimal fractional inhibitory concentration index (FICI), 0.375), A. baumannii (FICI, 0.5), and K. pneumoniae (FICI, 0.325), and, using ofloxacin, towards K. pneumoniae (FICI 0.5). Lack of hemolytic activity towards human red blood cells is an important benefit of the studied peptides when used at concentrations several times higher than those showing antimicrobial effects. The data obtained presume certain prospects for further investigations of proline-rich peptides, as well as their structural modifications, for the development of new combined drugs based on these compounds, in order to combat antibiotic-resistant microorganisms, e.g., medications for local applications, various components of medical devices, in particular, venous catheters, stents and wound dressings.

 

Search for novel prognostic criteria predicting successful in vitro fertilization remains a nonresolved problem at the present time. The aim of our study was to analyse a predictive role of IL-1α as an additional marker of pregnancy after in vitro fertilization (IVF). The study included 120 women with tubo-peritoneal infertility subjected to the IVF procedure. Retrospectively, two groups were formed of this cohort, dependent on efficiency of in vitro fertilization. Group I included 40 women with successful pregnancy whereas group II comprised 80 women with failed pregnancy. IL-1α concentrations in serum were detected by ELISA technique. A polymorphic rs1800587 marker at 5`UTR region has been amplified by PCR followed by Sanger sequencing. We have shown IL-1α hyperproduction in the women from group I. The women with effective IVF outcome exhibited positive correlation between IL-1α and luteinizing hormone, prolactin, progesterone, 17-hydroxyprogesterone, dehydroepiandrosterone sulfate levels. The women with ineffective in vitro fertilization have detected a negative correlation between IL-1α levels and anti-Muellerian hormone, a positive correlation of IL-1α with 17-hydroxyprogesterone. The women with T allele of the polymorphic rs1800587 marker at 5`UTR region have shown a 2.5-fold higher chance to become pregnant after IVF than the women carrying C allele (95% CI = [1.45-4.35], р = 0.0009). The women with T/T genotype exhibited a positive correlation between IL-1α and estradiol, testosterone; the subjects with heterozygous C/T genotype showed correlation with estradiol, and those harboring C/C genotype exhibited correlation with follicle-stimulating hormone. The revealed changes suggest a potential involvement of IL-1α into regulation of cyclic processes in the ovary including ovulation. Moreover, IL-1α may participate in formation of pro-inflammatory environment for successful blastocyst implantation.

 

Disorders in functioning of major regulatory systems in patients with somatic diseases required development of new and effective integrated approaches to their treatment and prevention. Effective electrotherapy (electrical sleep) is among these methods. Despite existence of studies proving high efficiency of electrical sleep results in therapeutic practice, some open questions remain concerning impact of this treatment upon neuropeptide-cytokine links of immune system, which is one of the most important effector of pathogenesis in cardiovascular diseases in young persons with hypertension from the group occupied with stressful jobs. In this connection, the aim of our study was to investigate the influence on electrical sleep upon neuropeptide-cytokine profile in arterial hypertension conditions accomplished by asthenic-neurotic disorders in young men from the group of stressful activities. The following treatment groups were formed: 1st (n = 12), antihypertensive therapy; 2nd (n = 10), complex therapeutic measures added to antihypertensive therapy plus minor tranquilizers; in the 3rd group (n = 12), electric sleep was performed. Neuropeptide-cytokine profile was investigated as serum contents of β-endorphin, proinflammatory (TNFα, IL-1β, IL-6) and anti-inflammatory (IL-4, IL-10) cytokines. In the course of the clinical and laboratory examination, the authors have found that electric sleep applied in a complex primary schedule, with antihypertensive drug treatment in patients with hypertension and asthenic-neurotic disorders proved to exert optimizing effect upon functioning of neuropeptide-cytokine pool of immune system, which manifested by stimulation of beta-endorphin production, a decrease via regulation of proinflammatory effectors (TNFα, IL-1β, IL-6), and increased anti-inflammatory cytokines (IL-4, IL-10).

 

Biological preparations (BP) obtained by gene engineering possess a special characteristic called immunogenicity, i.e. propensity of biological drugs to induce an undesired immune response associated with arising anti-drug antibodies. These antibodies can change BP pharmacokinetics and pharmacodynamics, and therapeutical efficacy. A significant proportion of hemodialysis patients with end-stage renal disease treated by recombinant erythropoietin (rEPO) have clinical features of resistance to such therapy. The aim of the study was to investigate whether anti-rEPO antibodies are associated with hemoglobin concentrations (Hb) and red blood cells counts (RBC) in hemodialysis patients, receiving long-term rEPO therapy. This research was performed at the Research Institute of Nephrology at the First St. Petersburg I.Pavlov State Medical University. Thirty-seven hemodialysis patients (pts) with end-stage renal disease and anemia treated with different rEPO formulations were included into the study. The patients were further divided into two groups: those with diminished and normal clinical response to rEPO therapy (DCR, n = 21 vs NCR group, n = 16, respectively). To determine threshold levels of antibodies to rEPO-beta (Roche, Switzerland) we tested blood serum samples of 35 healthy blood donors who never received rEPO in the past. Concentration of antibodies was measured by means of dot-blot method. The threshold antibody concentrations were defined by measurement of anti-rEPO concentrations in 2-fold stepwise dilutions (1:10 to 1:200) of blood sera from 35 healthy donors .The threshold value for rEPO-binding antibodies was 20.27 µg/ml (95 CI%±0.43). Antibodies to rEPO were found in 54 % of serum samples in the patients. Anti-rEPO antibodies concentrations correlated with mean values of hemoglobin and erythrocyte counts over a period of 12-months for the entire group of hemodialysis patients (r = -0.368, p = 0.025 and r = -0.336, p = 0.042 respectively). Concentration of anti-rEPO antibodies, and the mean weekly rEPO dose were significantly higher in DCR group, compared to NCR group (p = 0.0019). In conclusion, higher levels of anti-rEPO binding antibodies seem to be associated with decreased therapeutic response to the clinically applied rEPO formulations.

 

Mechanical jaundice (MJ) of malignant genesis is a clinical syndrome caused by obturation of bile ducts due to malignant tumor growth. It is characterized by specific pigmentation of skin, mucous membranes and sclerae due to increased accumulation of bilirubin in blood serum and other fluids and tissues of the organism. Various immune disorders are detected in patients with mechanical jaundice. Immune reactivity in mechanical jaundice is affected by toxic, infectious, or tumor factors. In connection with these findings, the aim of our study was to measure levels of some pro-inflammatory (IL-2, IL-18, TNFα, IFNγ) and antiinflammatory (IL-4 IL-10) cytokines in patients with malignant disease, depending on serum bilirubin levels. The study included 50 patients with MJ by malignant origin. A control group consisted of 125 practically healthy volunteers comparable with the study group by sex and age. Blood serum cytokine contents in patients and controls was determined by enzyme immunoassay using reagent kits produced by ZAO Vector-Best. Statistical evaluation of data was carried out by means of Statistica for Windows 8.0 and Microsoft Excel software. The levels of IL-2, TNFα, IFNγ and IL-18 were increased before surgery in the patients with mechanical jaundice of malignant genesis, while IL-4 and IL-10 values were reduced in comparison with healthy controls. Altered cytokine regulation in the MJ group manifests as increase in proinflammatory and immunostimulatory cytokine levels, along with decrease in anti-inflammatory cytokine amounts. Predominant feature of immune response may be regarded as prevalence of Th1-immune response when taking into account cytokine levels in blood serum of patients with MJ of malignant genesis. The features of cytokine regulation of patients with malignant MJ depend on the bilirubin content in blood serum. These changes were most frequent in patients with bilirubin level exceeding 60 μmol/L. In general, the quantitative indices of cytokines exhibited unidirectional changes in the groups of MJ patients with different bilirubin levels, except of IL-10. This parameter showed almost 6-fold increase in MJ patients with bilirubin level < 60 μmol/L against control group, and approximately 40 times with respect to the remaining 2 groups. This biological effect may occur due to malignant etiology of the MJ.