Orexins A and B are neuropeptides synthesized by a population of lateral hypothalamic neurons. Orexin’s physiological function consists mainly in regulating the sleep-wake cycle, eating behavior, and energy homeostasis. Axons of orexin-containing neurons are projected onto many structures of brain and spinal cord, thus providing a variety of their physiological effects. Moreover, the components of the orexinergic system are identified in various peripheral organs and tissues. The effects of orexins are mediated via two receptors (OX1R and OX2R) coupled with G-proteins (GPCRs). The classical signal transmission pathway through orexin receptors in neuronal cells includes an increase of the intracellular calcium as a result of the opening of TRPC membrane channels and IP3 endoplasmic reticulum (ER) channels. In addition to the classic orexin receptors signaling, there is an alternative pathway. Signal transmission through the alternative pathway leads to apoptosis of tumor cells. This pathway is probably due to the structural feature of orexin receptors compared to other GPCRs — the presence of a tyrosine-based immunoreceptor inhibition motif (ITIM). Such motifs are not limited to GPCRs, but are a hallmark of immuno-inhibiting receptors on lymphoid and myeloid cells. ITIM recruits either SHP1 and SHP2 protein tyrosine phosphatases or SHIP1 and SHIP2 inositol phosphatases, to mediate negative signal transduction. A further mechanism of the so-called orexin-induced apoptosis seems to include the p38/MAPK phosphorylation and the cytochrome c releasing from mitochondria, followed by activation of caspases 3 and 7 and cell death. It should be emphasized that this alternative pathway is present only in certain types of tumor cells. This review summarizes the available data on orexin-induced apoptosis of tumor cells from intestines, pancreas, stomach, prostate, endometrium, adrenal glands and glia, and also considers possible mechanisms for its implementation.

Here we review literature data on properties of a member of nuclear hormone receptors - peroxisome proliferator-activated receptor-α. It was shown that PPARα was expressed on different cells including dendritic cells, macrophages, B- and T-cells. We discuss structure of natural and synthetic ligands of PPARa, molecular and cellular mechanisms of PPARa regulation of lipid and carbohydrate cellular metabolism. PPARa activity in hepatocytes results in decrease of intracellular concentrations of lipid acids. This leads to reduction of VLDL cholesterol, increase in HDL-cholesterol and decrease in triglycerides in plasma of patients taking PPARα agonists. Modulation of PPARa activity may change multiple biological effects of glucocorticoids (GCS) and insulin resistance. It is assumed that PPARα agonists reduce side effects of GCS and at the same time enhance their anti-inflammatory activity due to transrepression of NF-kB. We analyzed the results of several randomized studies, meta-analyses devoted to assessment of efficacy and safety of PPARa agonist fenofibrate in patients with type 2 diabetes mellitus with high risk of micro- and macrovascular events. The studies showed good safety profile of monotherapy with fibrates as well as of their combinations with statins, ezetimibe. Fibrates reduced not only cardiovascular events but also overall mortality. We present the data on the role of PPARa in control of glucose and lipid metabolism in subpopulations of innate and adaptive immunity cells. The data show that glucose and lipid metabolism play an important role in the fate of cells of innate and adaptive immunity. The metabolic state of lymphocytes has dynamic nature and depends on their functional activity. Transition from dormant cells with relatively low metabolism rate to activated and proliferating cells is accompanied with increase of metabolic demands. This transition is supported with the switch from oxidative metabolism to anaerobic glycolysis (Warburg effect) after antigen recognition by T-cells and B-cells. It was shown that granulocytes, dendritic cells and M1 macrophages were dependent on glucose metabolism during their activation while M2 macrophages were dependent on fatty acids oxidation. In contrast with lymphocytes, activated myeloid cells do not proliferate well but still have increased glycolysis which is necessary for their effector function. It is stressed that modulation of immune cells metabolism via PPARα gives new opportunities to modulate intensity and duration of immune responses in chronic diseases. We analyze studies performed on animal models of some chronic diseases, human patients with rheumatoid arthritis and different phenotypes of osteoarthritis. Most of the studies showed clinical efficacy and pleiotropic effects of PPARα agonists: antiinflammatory, immunomodulating and lipid modulating, primarily reduction of triglycerides and increase in HDL-C. The presented literature data suggest efficacy of PPARα agonists against individual components of polypathies. This could reduce risk of polypharmacy and reduce direct treatment costs. It is not unlikely that the use of PPARα agonists in a patient with multimorbidity could prevent acquiring a new disease. These are merely suggestions and much effort and time is required to perform large-scale randomized controlled studies evaluating new indications for the use of PPARa agonists.

The embryo, being half an antigenically “foreign” organism, should elicit a maternal immune response. During evolution, however, the mechanisms ensuring successful development of pregnancy have been formed. In particular, among factors providing immune tolerance during pregnancy are some proteins associated with pregnancy. The pregnancy-specific β 1-glycoprotein (PSG, PSG1; SP1; PSβG1) is a dominant fetoplacental protein produced by cyto- and syncytiotrophoblast cells, and it exhibits immunosuppressive properties. Our team of authors possesses a patented method for obtaining native human PSG preparation from blood serum of pregnant women, a mixture of PSG1, PSG3, PSG7, PSG9, and their isoforms and precursors. This review presents an analysis of our results for the period from 2015 to 2020. We studied the immunoregu-latory effects of the obtained PSG preparation at concentrations comparable to those observed in pregnancy (1, 10, 100 |ag/mL). The study was performed with peripheral blood cells obtained from non-pregnant women. It was found that PSG significantly increased the percentage of adaptive Tregs in vitro, as well as expression of CTLA-4, GITR, and production of IL-10 by these cells. It has been shown that PSG has a stimulating effect upon indoleamine-2,3-dioxygenase (IDO) activity of peripheral blood monocytes. For Th17 cells, we have demonstrated that PSG can suppress differentiation and proliferation of these cells, along with reduced production of critical proinflammatory cytokines (IL-8, IL-10, IL-17, IFNγ, MCP-1, TNF α). As for the memory T cells, PSG suppressed CD25 expression and IL-2 production by them, along with simultaneous decreased expression of Gfi1, hnRNPLL genes, thus preventing the formation of the “mature” CD45R0 isoform. PSG has been shown to inhibit naive T cells’ conversion to the terminally differentiated effector subpopulation of helper T cells. When analyzing PSG effects upon cytokine profile of immunocompetent cells, it was found that the protein predominantly suppresses the Th1 cytokine production by the studied cell types, and regulates the Th2 cytokine production in divergent manner. The results obtained are consistent with general concept of immunosuppression during pregnancy. Thus, PSG could be one of the factors preventing formation and implementation of immune response to placental antigens.

Diagnostics of allergic diseases is a difficult issue, which requires distinct solutions, since this disorder is very common among the population. The overview focuses on complex diagnostics, including various methods that are most in demand at the present stage. The allergy diagnostics primarily include taking anamnesis, physical examination, instrumental and functional tests. Less often, the provocative tests are used, due to risk of severe adverse reactions. At the present stage, the role of laboratory diagnostics of allergies is growing, since, firstly, there is an increase in difficult-to-diagnose cases that require involvement of the entire medical armamentarium, and, secondly, the sensitivity and specificity of laboratory tests are improving. Among laboratory methods, the most significant are the assessment of the level of specific IgE, and the relatively new basophile activation test. The latter test is the main focus of the present review. It is functional and combines the advantages of provocative tests, during which conditions are created for the interaction of a potential allergen and effector cells of allergic inflammation, keeping safety for the patient. The data on the life cycle of basophils, their expression of membrane receptors, the content of granules, and ability to produce additional inflammatory mediators by the cells are presented. Participation of these cells in pathogenesis of allergic inflammation is being considered. Various mechanisms of basophil activation are discussed, both IgE-mediated and IgE-independent, which are similar in vivo and in vitro. Theoretical aspects of using the in vitro basophil activation test to estimate the hypersensitivity to a wide range of allergens are discussed. High sensitivity and specificity of the test for diagnosing allergies to food, household, pollen, insect and drug allergens are presented. Specific features of the basophil activation test related to the preanalytical, analytical and postanalytical stages of the study are highlighted. The factors influencing evaluation of this method are known. For example, difficulties in interpreting the test may arise while taking glucocorticosteroid hormones, in acute period of inflammation, with severe edema. The possibility of using this test to assess effectiveness of allergen-specific and anti-IgE therapy is being considered. A comparison of the basophil activation test, measurement of specific IgE and skin tests by various parameters related to performance and interpretation of results is carried out. Comprehensive diagnostics of allergic diseases, including usage of pathogenetically determined laboratory methods, will contribute to adequate treatment and, as a result, improve the health of the population.

Glioblastoma remains the most common and aggressive primary brain tumor today. Because of the neuroanatomical location of glioblastoma, conventional chemotherapy and radiation therapy have limited efficacy in patients with these tumors. Over the past decade, antitumor immunotherapy has become widespread among modern therapeutic approaches. The importance of immunotherapeutic methods lies in their ability to increase the effectiveness of cancer treatment and prevent relapses by enhancing the systemic and local immune response against tumor cells.

One of the most promising directions in modern immunotherapy is CAR-T therapy, or adoptive cell therapy using genetically modified T-lymphocytes. The functional advantage of CAR-T therapy is its ability to genetically modify lymphocytes, leading to their activation in vitro.

This review examines the key principles of CAR-T therapy and analyzes the published results of clinical trials for the treatment of glioblastoma using several modifications of CAR-T cells.

Increased incidence of ulcerative colitis (UC) is a prerequisite for searching new therapeutic approaches, primarily with an opportunity of site-directed impact on the colon lesion. UC pathogenesis is associated with dysregulated immune response, and limited effectiveness of basic therapy for the disorder. Vitamin D3 exhibits antioxidant, anti-inflammatory, immunomodulatory and other properties, it has been shown to be effective in some autoimmune diseases, thus prompting us to study its effect on immune status in UC. We aimed for studying the effect of vitamin D3, as a component of original rectal suppositories, upon clinical course and indexes of immune status in experimental UC. UC in rats was modeled with 3% oxazolone solution. The vitamin D3-containing suppositories (1500 IU) weighing 300 mg were administered per rectum every 12 hours for 6 days. On days 2, 4 and 6 of UC, the clinical features were assessed as well as blood leukocyte counts, numbers of CD3⁺, CD45RA⁺; absorbing and NBT-reducing abilities of blood neutrophils were determined; IgM, IgG, IL-6 and IL-8 concentrations in serum were also studied.

The DAI index increased in non-treated UC, along with raised neutrophil numbers in blood, their absorption and NBT-reducing activity was also increased, the total number of lymphocytes, including CD3⁺, CD45RA⁺ became higher, serum concentrations of IgM, IgG, IL-6, IL-8 increased. Local use of vitamin D3 in UC reduces DAI parameters, causes decrease in blood neutrophil counts, reducing and partially restoring absorptive and NBT-reducing abilities of neutrophils, decline of total lymphocyte counts in blood, partially restoring the CD3⁺ and CD45RA+ numbers, causing decline and partial restoration of serum IgM, IgG, IL-6, IL-8 concentrations. An association between clinical signs and indexes of immune status in UC was established under the conditions of vitamin D3 use. Conclusions: The protective effect of vitamin D3 in UC can be mediated by its antioxidant effect, changes in production of immunoregulatory cytokines, modulation of Th1-, Th2-, Th17-dependent reactions and Treg activity, being a pre-requisite for further studies to clarify the mechanism of vitamin D3 immunotropic action in UC,with an opportunity of using it in clinical practice.

Next generation sequencing is used to determine full-length sequences of HLA genes at the 4-field (allelic) resolution. The study was aimed at determining frequency and diversity of HLA alleles in a cohort of blood donors from the Registry of the National Research Center for Hematology who design ated themselves as Russians (including some not routinely typed variations in HLA gene regions). The studied population consisted of 1510 donors. HLA typing was performed by next generation sequencing. Libraries were performed with AllType NGS Amplification Kits (One Lambda, USA) and sequenced using MiSeq (Illumina, USA). Data analysis used the TypeStream Visual Software V2.0.0.68 (One Lambda, USA) and IPD-IMGT/HLA database 3.40.0.1. Arlequin 3.5 software was used for estimation of allele and haplotype frequencies, deviation from Hardy-Weinberg equilibrium. 82 HLA-A, 156 HLA-В and 85 HLA-С alleles were identified with four-field resolution. 45 HLA-DRB1 and 18 HLA-DQB1 alleles were identified with 2-3-field resolution. Considerable HLA diversity was found among the donors self-designated as Russians: the population had large numbers of distinct alleles at each HLA gene, high percentage of alleles (25-32% of HLA class I) were revealed only once. Sufficient numbers of new alleles were registered which are absent in the IPD-IMGT/HLA database. Considerable allelic diversity in Russian population is due to low-incidence alleles. Despite this diversity, the majority of HLA alleles detected at each locus were common. Significant HLA diversity of the donors was connected with a large number of alleles with rare occurrence. The novel alleles identified in our study differed from the known alleles by single nucleotide substitutions. The most common alleles at the four-field level were as follows: A*02:01:01:01 (27.1%), C*07:02:01:03 (13.1%), A*03:01:01:01 (13.0%), B*07:02:01:01 (13.0%), A*01:01:01:01 (11.6%) and C*07:01:01:01/16 (10.4%). The HLA alleles, which are common for Russian populations, are not always common or well-documented alleles in present catalogues. The data obtained in this study may be used as a reference sample for estimation of HLA allele frequencies in Russian population, for proper frequency evaluation of specific alleles when searching donors for allogeneic hematopoietic stem cell transplantation, as well as for association studies between HLA alleles and different diseases, and for research in population genetics.

The main histocompatibility complex — HLA system (Human Leukocyte Antigens) is among the most important genetic factors determining response of humans to infectious agents. The key role that HLA molecules play in immune response is to present the pathogen-derived peptides. Enormous molecular variability of HLA alleles in human populations have attracted close attention and became the basis for numerous studies aimed at evaluating the role of HLA genotypes for individual features of immune response to COVID-19, the new infection caused by SARS-CoV-2 β-coronavirus. Many studies have focused on search of specific alleles associated with both susceptibility and resistance to this disease. Separate HLA patterns were reported already. These patterns may be either universal to several populations, or rather peculiar, since distribution of HLA genes is different for various populations, depending on the living conditions, including specific protection from environmental pathogens. Therefore, it is evident that individual effects of HLA genotype upon occurrence and course of SARS-CoV-2 infection should be performed in comparison with the HLA distribution among the residents of appropriate region. The objective of this study was to compare the distribution of HLA-A*, B*, DRB1* allele groups, and to analyze the frequencies of HLA-AB-DRB1 haplotypes in subjects with COVID-19 (n = 138), compared with the control group presented by residents of the North-Western Russia (n = 1456). The most significant differences between COVID-19 patients compared with a group from control population were revealed for the groups of HLA-A* alleles: the frequencies of HLA-A*02 and HLA-A*26 were significantly reduced (39.86% versus 51.72%, χ² = 7,58, and 4.35% versus 9.07%, χ² = 4.17, respectively). At the same time, the frequency of HLA-A*29 was increased more than 2-fold (5.80% versus 2.47%, χ² = 4.03). This finding suggests that the allele groups A*02 and A*26 are associated with reduced likelihood of the disease, while A*29, is an apparent factor predisposing for susceptibility to the disease. It was found that occurrence of definite HLA haplotypes, including the A*02 allele group, is less common in persons who have undergone COVID-19, and are ranged at the 4^th, 7^th and 10^th positions in frequency, while in the population control group such HLA haplotypes took the 3rd, 4^th, 7^th and 8th places. Further evaluation of the HLA gene polymorphism will allow to understand the predetermined immunogenetic basis for susceptibility, as well as clinical severity of COVID-19.

It is well established that cohorts of individuals exposed to ionizing radiation exhibit increased risks for cardiovascular diseases. Currently, the role of immune system in pathogenesis of atherosclerosis is actively studied. Meanwhile, the immunomodulatory effects of irradiation in pathogenesis of atherosclerosis in the persons exposed to ionizing radiation still remain unclear. The aim of this research was to study the effect of ionizing radiation upon lymphocyte subpopulations involved in pathogenesis of atherosclerosis. The lymphocyte subpopulations were studied in peripheral blood of the workers chronically exposed to occupational combined radiation versus a control group. The study considered 72 workers of the Russian nuclear production facility, the Mayak Industrial Association (mean age of 72.1±10.9 years), and 72 control individuals (mean age of 70.7±9.2 years). All the workers were chronically exposed to combined radiation (external gamma-rays and internal alpha-particles). The mean cumulative dose absorbed by red bone marrow from external gamma-ray exposure was 0.750±0.699 Gy; the mean cumulative absorbed dose to red bone marrow from internal alpha-particles was 0.072±0.092 Gy. The relative and absolute numbers of lymphocyte subpopulations (total T-cells, T-helpers, T-cytotoxic, total B-cells, NK-cells and T-NK-cells) were detected by flow cytofluorometry. The absolute number of CD3⁺CD19⁺T-lymphocytes was significantly lower in the individuals exposed to chronic irradiation, compared with the controls (1658.8±694.3 x 10⁶/l and 1988.4±1045.4 x 10⁶/l, respectively). The relative number of CD3⁺CD4⁺T-helpers and CD3⁺CD8⁺T-cytotoxic lymphocytes was significantly higher in individuals exposed to chronic irradiation. Relative number of T-helpers in the main group was 42.4±8.8% vs 35.3±8.7% in controls; the relative number of T-cytotoxic lymphocytes was 27.6±9.5%, and 23.3±6.5%, respectively. A significant negative correlation was revealed between absolute number of T-lymphocytes and cumulative absorbed doses to bone marrow from external gamma irradiation (correlation quotient r = -0,53565, p = 0,000001) and internal alpha sources (r = -0.54804, p = 0.0000006). This correlation may indicate a relationship between these changes (decreased absolute numbers of T cells) and occupational exposure rates. The increased relative number of T-helpers and cytotoxic T-lymphocytes confirm an assumption that specific antigens may cause an enhanced immune response during the development of atherosclerosis in exposed individuals.

Scientific medical literature has accumulated a lot of data suggesting most important components of coronary heart disease pathogenesis and hypertension to be complex triggering processes of neuro-immune and neuro-endocrine interactions. Risk factors for cardiovascular diseases at the initial stages of atherosclerosis formation cause endothelial dysfunction and trigger a cascade of immune inflammation in coronary vessels, which is based on shifting immune response towards activation of lymphocytes, with predominance of cellular immune reactions. As a rule, it results in remodeling of the vascular wall under participation of proinflammatory cytokines, shifting blood lipid balance towards atherogenicity, destabilization of atherosclerotic plaque, development of thrombosis and acute coronary syndrome. In this respect, the aim of our work was to develop treatment methods that allow, under participation of endogenous immune regulators, to change the structure of pro-atherogenic links via their interactions at the initial stages of the atherosclerotic lesion formation in chronic coronary heart disease and hypertension.

To achieve this goal, 80 patients (men) were selected among the marine specialists of the ship crews serving in the Arctic latitudes and the Far North, with ischemic heart disease, stage 1 hypertension and astheno-neurotic disorders with anxiety and depressive manifestations. The groups of patients were formed as follows: Group 1 (n = 31, patients who received standard therapy with cardiotropic drugs; Group 2 (n = 29), subjects who underwent drug correction with weak tranquilizers as a part of standard cardiotropic therapy; Group 3 (n = 34), standard therapy accompanied by medical and psychological rehabilitation and digital psychophysiological therapy. Effectiveness of the treatment was studied by assessing the dynamics of parameters characterizing the neuropeptide-cytokine immune status, the markers used in the diagnostics of atherosclerosis, as well as paired relationships between them. The laboratory part of the work was represented by a set of diagnostic kits, including markers of atherosclerotic process, and test systems for determination of β-endorphin, proinflammatory cytokines (TNF α, IL-1 β, IL-6), and anti-inflammatory (IL-4, IL-10) spectrum.

We have found that the use of medical and psychological rehabilitation, along with digital psychophysiological therapy contributes to optimization of neuropeptide-cytokine interactions, thus showing efficiency of cardiotropic drugs usage. It seems to correct the relationships within proatherogenic structures of immune system and pathogenetic links involved in development of atherosclerotic process in polymorbid cardiovascular pathology from marine specialists with intense workloads.

Inflammation is among the factors promoting development of premature rupture of the membranes (PPROM). Upon the conditions of physiological immune imbalance in pregnancy, inflammation modifies its course and can even change the immune response. Appropriate indexes may be quantitative and functional. We used a marker of mitochondrial membrane potential (MPM, Ay) as an integral index of the functional state of immunocompetent blood cells (IBC) in 159 women who were examined at 8-14 weeks of gestation; they were observed up to 34-36 weeks. Of these cohort, 121 women were referred to a comparison group. The main group (n = 46) consisted of pregnant women with PPROM at the term of 28-33 weeks. The examination was carried out according to current medical standards, with informed consent, being approved by the Ethics committee at the Khabarovsk branch of Far Eastern Scientific Centre of Physiology and Pathology of Respiration — Research Institute of Maternity and Childhood Protection. Additionally, MPM and lymphocyte populations were determined by flow cytometry. The degree of disturbed energy supply in the IBC was based on the data of simultaneous determination of lymphocyte, granulocyte and monocyte numbers with reduced MPM values (application for invention No. 2020115963), thus revealing 3 degrees of energy deficiency: 1^st degree, monovariant IBC composition with reduced MPM; 2^nd degree, bivariant composition, 3^rd degree, total changes. A relative and absolute decrease in CD3 (72% vs 78% and 1624 vs 1980), CD8 (28% vs 33% and 651 vs 851), an increase in CD19 (14% vs 9% and 304 vs 219) were revealed in pregnant women with PPROM. When assessing MPM values in the IBC populations, a decreased proportion of women without energy deficiency from the 1^st to the 2^nd trimester (from 41% to 30%), due to the 3^rd degree of energy deficiency (from 17% to 26%) was detected. A shift of affected pools at the 2^nd degree of energy deficiency in favor of lymphocytic-granulocytic association (from 7% to 25%) from lymphocytic-monocytic compartment (from 73% to 50%) was found. From the 2nd to 3rd trimester, we have detected redistribution of granulocyte pools at the 1^st degree (0 to 8%) and from the lymphocytic-granulocytic association (25% and 5%) to monocytic-granulocytic (25% and 40%). In the group with PPROM, there was a decreased proportion of pregnant women without energy deficiency (13% and 27%), as well as with the 1^st and 2^nd degrees (17% vs 31% and 9% vs 17%), due to the 3^rd degree of energy deficiency (61% and 26 %), relative to the comparison group. The IBC pools of in the main group were redistributed at the 1^st degree in favor of granulocytes (25% and 8%), at the 2^nd, in favor of the lymphocytic-monocytic association (100% and 55%) from the granulocytic-monocytic (0% and 40%). Such imbalance of bioenergetic processes in the IBC can be an important factor of pathologically ongoing inflammation. These changes could be caused by both higher incidence of infections in such patients and by alloimmune interactions between mother and fetus. However, they may also determine the pathological course of inflammation. Preterm birth, which is usually caused by PPROM, is a multifactorial pathological condition. However, independent on specific triggers, the changes in energy supply of IBC, at least, may serve as a significant biomarker of probability for this disorder.

Isolated fractures of femur account for more than 10% of all road traffic injuries. Traumatic injury of femoral bone triggers a cascade of interrelated neuroendocrine reactions at systemic level, primarily at the hypothalamic-pituitary-adrenal axis, systemic response of immune system, initiated by release of tissue degradation products, cytokines and other mediators of damage into systemic blood circulation. Specific cellular reactions in response to traumatic injury to bone tissue include both innate and adaptive immune responses. In this regard, there is still scarce information on changes in blood lymphocyte subpopulations observed after closed isolated fracture of the femoral diaphysis at the middle third, before and after surgery. The aim of the present study was to evaluate the subpopulations of peripheral blood lymphocytes following closed isolated fracture of the femoral diaphysis with bone displacement in thecourse dynamics of surgical treatment, thus being required for studies in pathogenesis, development of diagnostic criteria and creating innovative treatment approaches. The study included 20 apparently healthy men and 36 men with closed isolated fracture of the femoral diaphysis of the middle third (32A and 32B, by AO/ASIF clinical classification, coded according to ICD-10 S72.3). The exclusion criteria were as follows: exacerbation of chronic comorbidities, diseases of lymphatic system and haematopoietic organs, oncological diseases, and evidence of osteoporosis. The spectrum of blood lymphocyte subsets was assessed on days 5, 7 (immediately after surgery) and on day 18 after closed isolated fracture of femoral diaphysis. We have found that, on the day 5 after IPBC along with leukocytosis in peripheral blood, the number of T-regulatory cells, cells with markers of early (CD25⁺) and late activation (HLA-DR⁺) proved to be increased, whereas representation of NK cells was decreased. On the day 7 after IPBC and immediately after surgery, leukocytosis persisted in blood, along with increased number of T-regulatory cells, CD3⁺ cells with early and late activation markers. On the day 18 after closed isolated fracture of the femoral diaphysis, the total numbers of leukocytes, T-lymphocytes, T-helpers, T-regulatory cells, T cells with an early activation marker are restored in peripheral blood, whereas the number of T-lymphocytes expressing HLA-DR⁺ molecules showed a significant increase.

Age-related changes in the oral mucosa immunity, including decreased contents of secretory immunoglobulins and antimicrobial peptides in saliva, along with changed balance of pro- and antiinflammatory cytokines, care risks for development of purulent-inflammatory or allergic diseases of the oral cavity. For example, denture stomatitis (DS) caused by Candida albicans occurs in about 30—70% of denture users. The purpose of this study was to assess the secretory immune state of oral mucous membranes in the patients with Candida-associated denture stomatitis. We examined 42 elderly patients (61-72 years old) with one-piece acrylic dentures for at least, 6 months (15 men and 27 women). Based on clinical and microbiological studies, the patients were divided into a group with DS (n = 24) and a group without DS (n = 18). The contents of secretory immunoglobulin A (sIgA) and proinflammatory cytokines was determined, i.e., interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-α (TNFα), and anti-inflammatory cytokines, e.g., receptor antagonist of interleukin-1 (RAIL), interleukin-4 (IL-4), interleukin-10 (IL-10), as well as antimicrobial peptides (cathelicidin LL-37, lactoferrin and alphadefensins 1-3 (HNP1-3). The sIgA levels in the salivary fluid of patients with DS (0.92 (0.80-1.26) g/l) were significantly lower (p < 0.05) than in patients without stomatitis (1.71 (1.23-2,13) g/l). In the group with advanced DS, a significant increase of IL-1β levels in saliva was observed, along with simultaneous decrease of IL-8 concentrations, compared to the other group, without differences in TNFα and IL-6 concentrations. Increased contents of IL-10 in saliva was also noted. It was shown that the concentrations of cathelicidin LL-37 in saliva of the DS patients was increased two-fold, whereas the contents of neutrophil-derived alpha-defensins (HNP 1-3) was decreased. Conclusions: The development of inflammation in denture stomatitis caused by usage of removable acrylic dentures associated with Candida albicans infection is characterized by functional insufficiency of the secretory immunity of the oral mucosa associated with decreased amounts of secretory immunoglobulin A and antimicrobial peptides of neutrophilic origin. Low levels of alpha-defensins may suggest a decrease in the functional activity of neutrophils in the elderly, thus leading to higher susceptibility to fungal infection of oral cavity.

Rapid progression of aesthetic medicine is a distinctive feature of present decade. In this area, leading position is taken by injection cosmetology, which is associated with an opportunity of pathogenetic approach to resolution of cosmetic problems primarily caused by skin aging. The most commonly used mesotherapy drugs, along with hyaluronic acid, are vitamins, amino acids, and microelements. Skin aging is associated with quantitative and functional changes in the local immune cell populations. In this case, it is rational to assume distinct effects of peptide complexes upon functional potential of immunocompetent cells. The aim of this study was to analyze time-dependent changes of some immune parameters after mesotherapy with a complex of hyaluronic acid and peptides. The observation group consisted of 26 women who received their course of mesotherapy for the first time. Objective instrumental evaluation of the effect with Aramo Smart Lite device showed that, after mesotherapy, the skin quality was significantly improving in comparison with pre-treatment conditions, due to decreased relief of skin creases and wrinkles, with a tendency for reduction of this effect six months later. When comparing the results of immunological testing in the patients after the course of treatment with the data before starting the therapy, we have found redistribution in the lymphoid cell populations, i.e., increased proportion of T-lymphocytes, decreased amounts of B cells, and CD16+ natural killers. Declined numbers of T-lymphocytes expressing early activation marker were associated with increased proportion of peripheral Treg lymphocytes. We have also detected activation of antibody production which manifested as increased levels of all major classes of serum immunoglobulins. Enhanced spontaneous oxidative activity of neutrophils was also noted. The results of immunological monitoring showed that, three months post-treatment, none of the quantitative and functional parameters of immunity was changed, as compared with the results obtained immediately after ending the mesotherapy. Six months later, however, all these indexes returned to their initial positions assessed before the cosmetic procedure. The changes in systemic immune response following mesotherapy with peptide complexes affect the mechanisms of both innate and acquired immunity, including differentiation of lymphocytes, their regulatory functions and activation potential, and provide modulation of effector reactions. Complete restoration of initial immune parameters is observed within six months.

Correct choice of nutrient media for culturing different types of cells in various applications is one of the most important aspects of modern biotechnology, since chemical composition of the culture media largely contains the necessary metabolites to support certain cells’ growth lines outside the body. Jurkat line of human leukemic T-lymphoblast-like cells (hereinafter Jurkat T-cells) is actively used for in vitro modeling of intracellular signaling and activation of normal blood T-lymphocytes mediated by the T-cell receptor/CD3/ CD4 complex in toxicological studies of immune and secretory responses, to test medicinal substances and ions. Also, Jurkat T-cells are widely used for ex vivo testing in immunology, oncology, toxicology, orthopedics, and traumatology. The existing standards and numerous studies are mainly based on short-term in vitro cultivation of Jurkat T-cells in RPMI 1640 nutrient medium. Meanwhile, the issues of long-term maintenance of the growth of Jurkat T-cells culture are poorly presented in the research literature. This study aimed for studying the activity of Jurkat T-cells over 7 to 14 days of in vitro culture and comparing the relative value of RPMI 1640 and αMEM media for the behavior of immunocompetent tumor cells. Using flow cytometry, multiplex analysis, and phase contrast Cell-IQ microscopy, the proportions of living cells and those dying by apoptosis and necrosis, secretion of cytokines and chemokines, and the dynamics of cell biomass propagation were studied. It was found that the αMEM medium in the complete nutrient medium, as compared with RPMI 1640, is more appropriate to in vitro promotion of cell viability (increased proportion of viable cells by 13.5% at the day 14), their secretory ability for 23 из 27 tested biomolecules, shortened adaptation time (на 32%) in culture before growth initiation, 5-fold increase of the Jurkat Т-cell cellularity by the day 7. Potential significance of the chemical components of nutrient media and secreted biomolecules for these results is discussed. As based on the results obtained, we concluded on superior properties of αMEM medium for long-term in vitro cultures of Jurkat T-cells. Consequently, the in vitro testing of medical devices intended for long-term contact with the body, including those for cancer patients, using Jurkat T-cell leukemia line in RPMI 1640 medium, may lead to wrong predictions on their biocompatibility and potential antitumor activity.

Our aim was to compare different immobilized erythropoietin (EPO) preparations for their ability to detect anti-EPO IgG antibodies in blood sera of EPO-treated patients with ELISA technique. 294 serum samples of the patients treated with erythropoietin were analyzed. 127 serum samples of patients who did not receive recombinant human EPO (rhEPO) were studied for comparative analysis. ELISA assay was performed, and different rhEPO drugs were immobilized on the anti-EPO monoclonal antibody-coated plates. Horseradish peroxidase-conjugated mouse monoclonal antibodies to human IgG, IgG1, and IgG4 was used for detection. The following drugs were studied: recombinant human erythropoietin rhEPO-beta (Shandong Kexing Bioproducts), European standard of erythropoietin BRP 3, commercial drugs Aranesp (Amgen Europe B.V.), Mircera (F. Hoffmann-La Roche Ltd.), Eprex (Johnson & Johnson LLC), Eralfon (Pharmaceutical Company Sotex). The sensitivity of the method was expressed as a positivity index (IP). IP calculated as the ratio of OD from tested sera to OD at the cut-off levels. The latter was assumed as a mean OD±SD for serum samples from EPO-naive patients. The results were evaluated as positive with IP > 1.1, and negative at IP < 0.9. Results in the range of 0.9 ≤ IP < 1.1 were considered as unidentified. Among the 294 samples, 32 specimens were evaluated as positive or unidentified for total IgG anti-EPO antibodies. The unidentified samples were detected in 1.0-1.7% of all cases. IgG1 subclass antibodies were found in 50-56.3% of patients and IgG4 subclass antibodies, in 43.850% of the patients. Mann—Whitney test showed a significant difference between the test samples compared to control group for all the ELISA modifications (p = 0.001). The Kruskal—Wallis test did not show significant differences between the IP results obtained with any of five immobilized EPO drugs (p = 0.05). The correlation quotient of IP was in the range of 0.99-0.96 for total IgG and > 0.98 for two subclasses of antibodies. Linear regression coefficients were close to one, thus indicating absence of significant differences in the sensitivity of the compared methods. This study indicate the opportunity of using the similar test systems to determine anti-EPO antibodies in the patients treated with various rhEPO drugs. Therefore, it is possible to develop a universal commercial test system to this purpose.