Vol 24, No 4 (2021)

Over last years, the world’s aging populations are rising rapidly, and the phenotypes of cognitive insufficiency, such as old age, depression and dementia, are increasing. Search for approaches to discrimination between such phenotypes is extremely relevant. Current studies present compelling evidence of the key role of immune system (its peripheral compartment), and the stress response system in physiological brain health. Therefore, assessment of complex interactions between immune and neuroendocrine systems may be an effective way to differentiate between depression and early stages of dementia in elderly people. Our purpose was to reveal peripheral molecular messages, e.g., cytokines and stress hormones, in the context of cognitive impairment phenotypes: healthy old age/old age depression/dementia. Eighty elderly people were included into groups as follows: “Healthy ageing”, “Dementia”, “Depression”. Levels of certain cytokines: IL-6, IL-1β, TNFα, IFNγ, IL-10, and stress hormones (cortisol, ACTH, dopamine, noradrenaline, and adrenaline) were determined in blood plasma by ELISA. The intergroup differences were evaluated by the Kruskal-Wallis test with Conover-Inman post-hoc pairwise comparisons. For differential diagnostics between the groups of elderly people with varying grades of cognitive impairment, we used linear canonical discriminant analysis performed on the ranks. It has been shown that cognitive insufficiency phenotypes—old age depression and dementia—differ from the healthy ageing phenotype with their high peripheral levels of TNFα cytokine and low levels of IL-1β. The differences between depression in elderly and dementia included lower level of IL-10 in depression (lower than in “Healthy ageing”), and high IL-6 in dementia (compared to “Healthy ageing”). Evaluation of the hypothalamic-pituitary and sympatho-adreno-medullary axes hormones showed hyporesponsiveness of hypothalamic-pituitary axis, regardless of cognitive insufficiency phenotypes, along with activation of sympatho-adreno-medullary axis, i.e., high dopamine level in old age depression with dementia, and high adrenaline level in dementia, than in depression of elderly phenotype and healthy ageing. Such significant differences in the levels of molecular messages, i.e., cytokines and stress hormones among the old age person groups, enabled diagnostic efficacy of 87.5% to differentiate cognitive phenotypes of aging: healthy ageing, old age depression, and dementia.

The article presents the results of evaluating growth factors, pro – and anti-inflammatory cytokine production by multipotent mesenchymal stem cell cultures under the conditions of co-cultivation with immuno-isolated beta-cells of the pancreas. β-cell transplantation is a minimally invasive therapeutic approach (compared to transplantation of entire pancreas), and it provides better metabolic control with respect to insulin administration. However, when transplanting β-cells, there is always a risk of immune rejection of the grafted cells. It is generally recognized that encapsulation is an effective means of immunological protection against the recipient’s immune system during transplantation. Regulation of the autoimmune response to transplanted cells is crucial for the treatment of type I diabetes mellitus. In recent years, along with replacement of islet cells, much attention has been paid to the use of multipotent mesenchymal stem cells with immunomodulatory and/or immunosuppressive properties, aimed for the correction of diabetes mellitus. Either in vitro and in vivo, they impact not only T-lymphocytes, but also B-lymphocytes, dendritic and NK-cells. Mesenchymal stem cells are able to inhibit proliferation of immune cells and reduce their secretion of inflammatory cytokines, acting as auxiliary cells to improve the survival of islets in the early post-transplant phase. Combined transplantation of multipotent mesenchymal stem cells and pancreatic β-cells is a promising approach to the treatment of type I diabetes mellitus. Deeper study of the mechanisms that cause their cytoprotective effect upon the transplant may be helpful for implementation of this therapeutic approach and improve its efficiency. In our study, a 1% solution of low-viscosity sodium alginate with addition of saline solution (0.9% sodium chloride) was used to create immuno-insulating scaffolds, and a 2.2% BaCl2 solution was added for polymerization. Decreased production of proinflammatory cytokines (TNFα, IL-12, IL-5) and growth factor (GM-CSF) was registered in co-cultures of β-cells with mesenchymal stem cells of bone marrow origin, and those obtained from subcutaneous adipose tissue. Anti-inflammatory activity was more pronounced in adipose stem cells and their immunomodulatory effects were shown via changes of their cytokine-producing activity. Hence, the multipotent mesenchymal stem cells obtained from adipose tissue and bone marrow have shown to exert cytoprotective effect upon pancreatic beta-cells by shifting the cytokine-producing activity towards an antiinflammatory profile.

Studies in HIV infection remain an important issue for modern medicine, which, becomes controlled due to widespread usage of antiretroviral therapy. At the same time, however, it cannot be cured completely, and there is a number of “white spots” in understanding immunopathogenesis of disorders complicating this disease. The in-depth studies of interferon system, in particular from the lambda family, are desirable, because of their antiviral activity in HIV-infected patients. The aim of our study was to evaluate the content of interferons-lambda: IFNλ1 (IL-29) and IFNλ3 (IL-28B) in patients with HIV infection.
Blood serum of 120 patients with HIV infection (average age 49.7±6.2 years) who were treated in the outpatient setting at the Center for AIDS Prevention and Control of Regional Clinical Hospital No. 2 in Vladivostok was subjected to laboratory testing. HIV infection 4A was the main clinical diagnosis in all the patients, i.e., the stage of secondary diseases, remission phase on the background of antiretroviral therapy (ARVT). In 52 patients, chronic viral hepatitis C was found as a concomitant disease. The content of IFNλ1 (IL-29) and IFNλ3 (IL-28B) in venous blood serum was determined by ELISA technique using a “Multiscan” analyzer. The ELISA reagents were from R&D systems, catalog numbers DY5259; DY7246.
In the groups of patients with HIV infection, both with and without viral hepatitis C, the levels of IFNλ1 (IL-29) and IFNλ3 (IL-28B) were significantly reduced in comparison with control group. When comparing the IFN values between the groups, a more pronounced decrease in IFNλ1 (IL-29) was revealed among the patients with viral hepatitis C. When analyzing the level of IFNλ3 (IL-28B), an opposite pattern was observed, i.e., its values in the patients with HIV infection and viral hepatitis C were higher than in the group without hepatitis, but still did not reach appropriate values of the control group. Based on the data on IFNλ3 (IL-28B) antiviral effect upon HIV transmission via macrophages One may assume that induction and maintenance of higher type 3 interferon levels can favorably affect the course of HIV infection. The registered changes in IFNλ1 (IL-29) and IFNλ3 (IL-28B) levels in the patients with HIV and HCV suggest some viral effect upon innate immunity characterized by multidirectional changes, depending on presence or absence of hepatitis C virus. The studies of changes in innate immunity and role of type 3 interferons may extend our knowledge on the interaction between the human body and HIV, and will promote preventive measures and rehabilitation in the patients with HIV infection.

Sepsis is nearly always associated with some type of haemostatic disorder. The factors that play main causal role in pathogenesis of these processes are pro-inflammatory cytokines, vascular endothelium, platelets, leukocytes, and tissue factor (TF) expressed on these cells, which is always in an active state. Given a potential relationship between the blood clotting and pathophysiology of sepsis, TF may be considered a biomarker for early diagnosis, risk stratification, and prognosis of disease outcome in sepsis. Objective – to study quantitative content (CD14+CD142+) and the levels of TF expression on monocytes in the patients with sepsis, to analyze the dependence of these parameters on the severity of multiple organ dysfunction according to the SOFA scale, and disease outcomes.
67 patients with sepsis were examined. The severity of multiple organ dysfunction/failure was assessed by means of the SOFA score (Sepsis-related Organ Failure Assessments, Sequential Organ Failure Assessment). All the patients were divided in 2 groups based on the severity of their condition and extent of organ failure. Group 1 (n = 30) included the patients diagnosed with sepsis and severe organ dysfunction of < 6 points on the SOFA scale; Group 2 (n = 37) consisted of the patients with sepsis and organ dysfunction of > 6 points according to the SOFA scores. Blood sampling from patients was made within initial 48 hours after admission and diagnosis. Quantitative content (CD14+CD142+) and the level of expression of tissue factor on monocytes were investigated by flow cytometry. We have found that the content of (CD14+CD142+) cells was significantly higher in patients with sepsis than in healthy individuals (6.03±1.05% vs 0.24±0.02%, p = 0.001), being higher in more severe organ dysfunction (SOFA) vs less severe cases (SOFA) (6.50±0.98% versus 4.42±0.36%, p = 0.05). High level of TF expression on monocytes showed a direct correlation (r > 0.71; p = 0.05) with severity of organ dysfunction (SOFA), and it was associated (p = 0.004) with lethal outcome of the disorder. These results suggest that expression of tissue factor on monocytes can serve as a biomarker reflecting the degree of systemic inflammation in sepsis, thus being a criterion for predicting clinical severity and outcome of the disease in patients with sepsis.

The immune system responsible for genetic stability of internal environment, with its exceptional sensitivity, may provide biological indexes which reflect impact of various environmental (biotic and abiotic) factors, i.e., serve as an indicator system in the areas of environmental well-being and distress. A series of studies on the influence of climatic and environmental factors on immune reactivity states demonstrated that the immune system is also sensitive to both natural and industrial factors (chemical and physical). Apparently, this is due to formation of a population-based immunological reactivity adapted for specific environmental conditions. However, the distribution patterns of immunogram variability by geographical zones are still unclear. The article analyzes prevalence and structure of allergic disorders in children of 1-3 and 4-7 y.o. (early and first childhood)living in different climatic and ecological areas of Kaliningrad region (coastal and continental zones, “clean” and “dirty” cities, different for anthropogenic burden and environmental situation). The following cities were involved into the study: Svetlogorsk, Svetly, Kaliningrad, Sovetsk, Gusev and Neman. There were 3,321 children examined. Clinical and demographic data were taken from the following sources: “History of the child’s development” (Form No. 112/y), “Outpatient medical record card” (Form No. 025/y) and “Diagnostic card of immunological deficiency in children”, developed at the Institute of Immunology (Federal Medical&Biological Agency of Russia) which contained the patient’s data, seasonality of increased morbidity, vaccination history, acute infections). Disorders and reactions attributable to allergic syndrome were divided into three groups: skin diseases, respiratory system diseases, adverse response to various antigens. The revealed differences demonstrate high sensitivity of immune system to climatic and environmental factors acting at the sub-threshold level. We believe that the child’s immune system responds even to minor environmental factors, which, acting in combined manner, cause its functional shifts. In turn, these changes manifest at the level of immunobiological reactivity and, accordingly, influence clinical course and symptoms of the allergic disorders.

The formation of immunity in the population to various variants of the COVID-19 pathogen is one of the most important problems for the world community. The aim of the study was to compare the dynamics of M-CSF and VEGF-A, IL-34 in the blood serum of patients with essential hypertension (EH) stage II, depending on the type of immunity formed (post-infectious, post-vaccination, “hybrid”) to analyze changes of M-CSF-mediated mechanisms of hypertension development. During the work, 2 groups of patients were formed: group 1-patients with stage II EH and SARS CoV-2 infection without pneumonia in the anamnesis, vaccination 6 months after laboratory recovery, group 2 – patients with stage II EH without COVID-19 in the anamnesis, vaccination during the follow-up period. Determination of M-CSF, IL-34, VEGF-A, IgG level to SARS-CoV-2 was determined using an enzyme-linked immunosorbent assay. The formation of post-infectious immunity in patients with stage II EH, despite the mild course of COVID-19, is accompanied by a long-term (up to 6 months) pathophysiologically significant increase in the serum level of M-CSF (p < 0.001) with a decrease in IL-34 (p < 0.001). Analysis of the dynamics of changes in M-CSF, IL-34, VEGF-A in the postvaccination period in the blood serum of patients with stage II EH with COVID-19 in the anamnesis (“hybrid” immunity), determined the absence (p < 0.05) of changes in the levels of M-CSF, VEGF-A after the first component of “SPUTNIK V” against the background of an increase of the level of IgG to SARS-CoV-2. 21 days after the second component of the vaccine, an increase of M-CSF was detected when compared with both pre-vaccination indicators and data 21 days after the introduction of the first component of the vaccine (p < 0.001). Comparing the dynamics of the of M-CSF, IL-34 and VEGF-A in patients with stage II EH without COVID-19 in the post-vaccination period with the data on the formation of “hybrid” immunity, an increase in M-CSF was recorded 21 days after the introduction of the first and second components against the background of a decrease in IL-34, but with the restoration of pre-vaccination concentrations in 100% of patients by day 180 with comparable immunogenicity after 180 days. In patients with EH II, the pathogenetic “summation” of the pre-infectious imbalance of cytokine regulation and postcovid changes is important, which in a number of patients may be the reason for the prolongation of the stabilization of the balance of the M-CSF-IL-34-VEGF-A system in the post-vaccination period during the formation of “hybrid” immunity.

Immunization with whole-cell adsorbed diphtheria-tetanus-pertussis vaccine (DTP) can cause various undesirable effects. The most common complications are febrile seizures, neuromyalgic syndrome and, in more severe cases, various encephalopathies. The listed complications are quite dangerous, especially in childhood, when primary DTP immunization is carried out. Many studies indicate that the development of these pathological processes is often associated with the action of various cytokines produced in response to vaccination. There are specific pro- and anti-inflammatory cytokines, the high levels of which are associated with the development of neurological complications after DTP vaccination. For example, IL-6 and IL-1 are often associated with the development of febrile seizures and encephalopathies. On the other hand, there are clinical data indicating a decrease in the incidence of complications after concomitant administration of vaccines. Thus, it is of particular interest to study the cytokine profile after the combined administration of DTP with another vaccine, which in some cases leads to a decrease in the number of complications and better tolerance of the vaccine. The vaccine against polio infection is currently one of the safest, but its effect on the level of cytokines is extremely poorly understood. Moreover, due to the fact that these drugs have been used for a long time and the interest in them is not as high as before, the number of new experimental works specifically on the cytokine profiles of many vaccines is limited. Basically, all existing work is aimed at studying various pathological processes associated with the introduction of a vaccine preparation. This leads to the fact that the mechanism of the formation of the immune response remains not fully understood. The aim of this work was to study the effect of combined vaccine administration on the cytokine profile. Results were obtained for the following cytokines: IL-2, RANTES, Eotaxin, MIP-1β, IL-12p40, IL-4, IL-6, IL-1α, and G-CSF determined in murine serum samples after combined administration of DTP and polio vaccine to the experimental animals. The cytokine profile was determined using Bio-Rad MAGPIX fluorescence reader. The study revealed and described the patterns of changes in the cytokine profile, both with the administration of the poliomyelitis vaccine alone, or in combination with the DTP vaccine. The results obtained in this work may be further used for more detailed studies on the mechanism of the immune response formation upon combined administration of vaccines and further improvement of existing drugs.

Rheumatoid arthritis (RA) is a multifactorial disease, with genetic component based on intergenic interactions leading to the formation of gene networks. The current trend in RA immunogenetic studies is to assess the gene-to-gene interactions. Among possible genetic factors contributing to RA development, the genes of main histocompatibility complex (HLA class II) play a fundamental role. TNFA gene is among possible candidate genes providing susceptibility to this disorder and contributing to its immune pathogenesis. The special location of this gene suggests arrangement of extended TNFA – HLA haplotypes. This work analyzed the distribution features of two-locus SNP haplotypes (TNFA and HLA DRB1) for their association with rheumatoid arthritis in Russians. The following methods were used: DNA isolation, PCR-based genotyping, RFLP analysis with electrophoretic detection. Calculation of two-locus haplotypes frequencies and linkage disequilibrium (D’; χ2; p) was carried out using the EM algorithm in the Arlequin ver 3.5 program. Comparison of paired samples was carried out using standard immunogenetic criteria. The significance level was < 0.05. Analysis of the data showed that the two-locus haplotypes -1031T/C and -863C/A TNFA were not associated with predisposal for rheumatoid arthritis in Russian population sample. The haplotypes associated with predisposition for RA were TNFA -863*a – HLA DRB1*03, TNFA -1031*t – HLA DRB1*04, TNFA -1031*t – HLA DRB1*04. Meanwhile, TNFA -1031*t – HLA DRB1*15; TNFA -1031*t -HLA DRB1*11 proved to be protective haplotypes. Our study showed that, in addition to individual HLA II alleles, the predisposal or resistance to rheumatoid arthritis may be promoted by haplotypes of rare SNPs at positions -1031, -863 C/A of TNFA gene, and HLA DRB1.