Vol 27, No 2 (2024)

The main stages of maturation of antigen-specific B cells occur in the germinal centers of the lymph nodes. During the process of differentiation, a decision is made on which path the B cells will take to develop further. They will either turn into short-lived plasmablasts or memory B cells or plasma cells. The relationship between these processes is very important for the development of a productive humoral immune response. The goal of the work was to create a system that is capable of simulating ex vivo processes occurring in germinal centers. We used primary B cells from human peripheral blood as starting material. B lymphocytes were stimulated in vitro using feeder cells carrying CD40L molecules and recombinant IL-21. Upon IL-21/CD40L stimulation, B lymphocytes changed their morphology, surface phenotype, and functional activity. After active expansion for 10 days, further cell growth stopped, and after some time they died. To generate stably proliferating B cells, we used lentiviral transduction of IL-21/CD40L stimulated IgM⁺ B cells. For this purpose, lentivirus preparations were obtained that carried a cassette consisting of the BCL6 and BCL2L1 genes, separated by a sequence encoding the self-cutting peptide P2A, as well as a GFP reporter gene separated from the target genes by an IRES element. The cassette used ensured the synthesis of the Bcl-6 transcription factor and the Bcl-XL protein in target cells. The Bcl-6 repressor prevented B cells from undergoing terminal differentiation and becoming plasma cells, and the Bcl-XL protein had an anti-apoptotic effect. Transduced B cells proliferated for more than a month and maintained a plasmablast phenotype. Forty-two days after the start of stimulation, transduced B cells remained GFP-positive, coexpressed CD27 and CD38 antigens, carried surface CD20 and IgM, intracellular Bcl-6, Bcl-XL and IgM, retained IgM secretion, but remained negative for surface and intracellular IgG. The proven stimulation system will allow us to simulate key aspects of B cell development in germinal centers to study the formation of B cell memory, which will ultimately facilitate the development of effective vaccines.

Lipopolysaccharide (LPS)-induced lung injury is the most commonly used mouse model of acute lung inflammation that simulates the development of respiratory distress syndrome in humans. The effects of acute LPS-induced airway inflammation are well studied and associated with the neutrophil accumulation in bronchoalveolar lavage fluid (BALF), local and systemic production of proinflammatory cytokines and narrowing of the airways. Recent studies demonstrated the presence of pulmonary fibrosis characterized by increased fibroblast proliferation and excess extracellular matrix deposition in late phase of acute lung inflammation caused by LPS exposure. This work describes an experimental model of acute lung injury induced by a single aerosol injection of LPS as a reproducible in vivo model of pulmonary fibrosis. To induce lung injury, C57BL/6 mice were placed in a chamber and exposed to an aerosol containing 10 mg of LPS using an Aeroneb Lab Nebulizer delivery system. We found that 5 weeks after a single nebulized LPS administration, mice have increased production of IL-6 in BALF. Although the frequency of neutrophils was not altered, there was a decrease in the percentage of alveolar macrophages at 5 weeks after LPS exposure, indicating continued lung inflammation. Several weeks after aerosolized LPS challenge, IL-10 production in BALF was increased, as well as expression of Tgfb1, Col1a1, Il13 and Acta2, and collagen deposition in lung tissue compared to mice with acute lung inflammation.

Thus, the single nebulized LPS administration represents a relevant, reproducible and physiologic model in mice allowing to investigate the mechanisms of pulmonary fibrosis development and help in the search for new therapeutic agents and approaches.

Mast cells, due to the mediators contained in them, are active participants in various processes occurring in the body. The reaction of avidin-positive mast cells of the liver to the intake of silicon with drinking water at a concentration of 20 mg/L for nine months was studied. The experiment was conducted on laboratory nonlinear male rats, which were divided into two groups: the control group received bottled drinking water with a silicon concentration of 10 mg/L; the experimental group received the same water, but with the addition of sodium metasilicate nine-hydrate, which was used to adjust the total concentration of silicon in drinking water to 20 mg/L. The mass concentration of silicon in the water of the control and experimental groups was determined using an inductively coupled plasma emission spectrometer 5110 ICP-OES. After nine months, the animals were sacrificed, the liver was extracted and fixed in 10% neutral formalin, processed and embedded in paraffin. After deparaffinization, 6-ìm-thick liver sections were incubated for 30 minutes with green fluorescent-labeled avidin (Avidin, Alexa Fluor^® 488 conjugate, Invitrogen, Germany). The preparations were analyzed under a fluorescence microscope at an excitation light wavelength of 495 nm. According to the results of the study, an increase in the number of avidin-positive mast cells due to weakly fluorescent ones was found in the liver of rats of the experimental group, as well as an increase in the median of their area and fluorescence intensity. It was revealed that the change in the median of area of avidin-positive mast cells in the liver of rats of the experimental group is due to a decrease in the number of small cells and an increase in the number of medium and large cells. Positive strong and positive average correlations were established between the cell area and its intensity in the control and experimental groups, respectively.

Thus, the study made it possible to expand the data on the effect of water-soluble silicon on the liver and indirectly suggest the occurrence of inflammatory processes in the organ under study.

Holothurians are among the most capable of regenerating animals. Their phagocytes are analogues of macrophages, but the mechanisms of their participation in wound healing have not been studied. The aim of the work is to determine the effect of individual protein components isolated from the coelomic fluid of holothurians with superficial damage to the body wall on the functional activity of the two types of phagocytes (P1 and P2) in holothurian Eupentacta fraudatrix.

Protein components of the coelomic fluid of holothurians with superficial wounds of the body wall were analyzed and collected by gel permeation chromatography. We used proteins whose content changed significantly during the wound healing period in preliminary experiments. Two proteins and a peptide were added to phagocytes, isolated by gradient centrifugation, simultaneously with the thermostable toxin of Yersinia pseudotuberculosis and incubated for 24 h. The production of superoxide anion radical was determined by the reduction of nitroblue tetrazolium (NBT) using a colorimetric method. To assess the specificity of the proteins, bovine serum albumin (BSA) was used as an internal control. In one-day in vivo experiments, proteins were administered to wounded animals.

Two proteins, when compared with the effect of BSA, did not show a specific effect on the production of reactive oxygen species in phagocytes in vitro. However, a comparison of the effect of the peptide on the oxidative activity and viability of phagocytes with those of BSA revealed the specificity of its action. The peptide reduced the oxidative activity of P1 phagocytes and increased it in P2 phagocytes in a direct concentration-dependent manner. One of the two proteins being administered to wounded holothurians caused a multidirectional concentration-dependent effect on the oxidative activity of the two types of phagocytes, with a predominant suppression of the P1 phenotype activity. The ability of protein components to cause a shift in the ratio of oxidative activity of the two types of phagocytes towards the preferential activation of P2 phagocytes, which are considered as functional analogues of M2 macrophages, suggests that they act as anti-inflammatory agents.

Morphological changes in the liver during infection with Opisthorchis felineus are characterized by significant structural changes. It has been proven that O. felineus modifies immune reactivity by increasing the synthesis of IL-10 and TGF-β and decreasing the levels of IL-4 and IL-5. The experiment was carried out on 15 Mesocricetus auratus aged from 4 to 6 weeks. The animals were divided into 3 equal groups. The duration of observation was 24 days. Live metacercariae of O. felineus were isolated from infected cyprinid fish. Infection was carried out by oral administration of 50 metacercariae. Animals in the experimental groups were injected intraperitoneally with 1 ml of the test supernatants. Histological preparations were examined by video microscopy with morphometry. It has been established that in the acute phase of opisthorchiasis, general infiltration of the portal tract area predominates. The cellular composition of infiltrates is characterized by the presence of lymphocytes, macrophages, fibroblasts, epithelioid cells and foreign body cells. The described cell changes are associated with a local decrease in oxygen access to hepatocytes in the peripheral zone of the lobule due to their compression by fibrous tissue. In the liver of the experimental groups, only tissue-specific changes were noted, and the damage to hepatocytes was the least pronounced in the group with the introduction of B. bifidum supernatant. The cytokine profile in the control group corresponded to the Th1 variant. In experimental group 1, when using the supernatant of B. bifidum, in comparison with the control, the ability of lymphocytes to produce the main pro-inflammatory cytokine, TNFα, decreased 4 times; a suppressive effect was also observed in relation to IL-1β, IL-2, IL-6, IL-8, and IFNγ. Inhibition of IL-10 production was observed, while the level of IL-4 and IL-5 increased in comparison with the control. When using the supernatant of chicken embryo cells, we observed similar results to group 1: a suppressive effect on pro-inflammatory and activation of the production of anti-inflammatory cytokines. IL-17 levels decreased in both experimental groups. Activation of the production of anti-inflammatory cytokines in experimental groups modulates the immune response towards Th2, which helps block inflammatory effects.

The main method of treating pain syndrome is the use of morphine and its analogues, which have a wide range of side effects. In this regard, many modern studies are aimed at finding safer analgesic compounds. Currently, among morphine analogues, much attention is paid to endogenous opioid peptides, which, along with an analgesic effect, have pronounced immunoregulatory properties. It is now known that the functional activity of immune cells is ensured by metabolism. This process supplies cells with the energy necessary for activation, differentiation, proliferation, apoptosis, etc. Glucose metabolism plays a key role in ensuring the functions of immune cells. The aim of this work is to evaluate the effect of endomorphins-1, 2 on the characteristics of glucose uptake by T and B lymphocytes in vivo. The subjects of the study were white male mice, peptides were administered to mice intraperitoneally at a dose of 100 ìg/kg, and glucose uptake by cells was assessed using fluorescent analogues of glucose (2-NBDG). It was found that endomorphin-1 did not affect at the intensity of glucose consumption in both T and B cells. Administration of endomorphin-2, on the contrary, led to a significant increase in glucose uptake in T lymphocytes. However, the level of glucose consumption in B cells after administration of endomorphin-2 did not change significantly. In a study of two subsets of T lymphocytes, it was noted that endomorphin-2 leads to an increase in glucose uptake in both CD4⁺ and CD4^-T cells. Administration of endomorphin-1 had no significant effect on the level of uptake of this substrate in both subsets of T lymphocytes. Proliferating B lymphocytes increased glucose consumption in the presence of LPS after administration of endomorphin-2. Both endomorphins did not have a significant effect on glucose consumption in proliferating CD4⁺ and CD4^-T cells.

Thus, endomorphin-1, unlike endomorphin-2, does not have a significant effect on glucose metabolism in T and B cells. Taking into account the role of glycolysis in the functioning of immune cells and inflammation, it can be concluded that the use of endomorphin-1 may be associated with a lower risk of immune system-related side effects.

Peripheral blood CD4⁺T lymphocytes are heterogenous, including naive, central memory, effector memory, and terminally differentiated effector cells. Each subset performs different functions and possesses unique metabolic properties. Mitochondria are vital organelles of CD4⁺T lymphocytes, playing critical roles in metabolism, energy and active oxygen species production, cellular respiration, proliferation, differentiation, and apoptosis. The use of mitochondrial-selective fluorescent dyes in combination with labeled monoclonal antibodies is a relatively accessible and simple way to study a range of mitochondrial parameters in CD4⁺T cells of varying maturity by flow cytometry. The aim of this study was to investigate mitochondrial indices in different CD4⁺T lymphocyte subsets. We obtained mononuclear cells from peripheral blood of nine relatively healthy volunteers. By flow cytometry using commercial fluorescent dyes MitoTracker Green FM and MitoTracker Deep Red FM, we determined the mass and membrane potential of mitochondria in the total pool of CD4⁺T lymphocytes and in their subsets: naive (CD45R0^-CCR7⁺), central memory (CD45R0⁺CCR7⁺), effector memory (CD45R0⁺CCR7^-), and terminally differentiated effectors (CD45R0^-CCR7^-). We show that in healthy individuals, central and effector memory CD4⁺T lymphocytes compared to naive cells have increased mitochondrial mass and membrane potential. The mass of organelles in functionally different memory CD4⁺T cell subsets vary significantly: it is lower in central memory lymphocytes than in effector memory cells. Nevertheless, two subsets have similar mitochondrial membrane potential. Terminally differentiated effectors differ from other CD4⁺T lymphocyte subsets in unique characteristics of mitochondria: despite high mass, they have a reduced membrane potential. This feature may be linked to cells being prepared for programmed cell death during the terminal differentiation stage.

A significant increase in the prevalence of diseases linked with IgE production can be seen in recent years, but the question about the role of TLR receptors in this process remains controversial. According to the hygiene hypothesis, the decrease of the contact of the individual with pathogens that contain PRR receptor ligands in the recent years leads to the development of allergic diseases. The aim of this work was to investigate whether TLR4 and NLRP3 receptor activation contributes to allergen-specific antibody formation.

BALB/c mice were immunized according to two different protocols. In the first one, OVA antigen was administered in 0.1 µg dose 2-3 times a week for 6 weeks by subcutaneous route. In the second one, OVA was administered in 0.3 µg dose intranasally in combination with 4 ng of benzo(a)pyrene (BaP) 2 times a week for 8 weeks. In both cases, TLR4 and NLRP3 receptor inhibitors, namely TLR4-IN-C34 in 1 mg/kg dose and CY- 09 in 20 mg/kg dose respectively were also administered to the some of the mice. Specific antibody production was determined by ELISA.

Immunization of mice with TLR4-IN-C34 significantly (p < 0.01) amplify IgE production (about 2.5 times in comparison with control group), but has no effect on specific IgG1 production in subcutaneous model. Specific IgE titers in the control group immunized without small molecule inhibitor and in the TLR4-IN-C34 group were (3±0.6) × 10³ and (8±2) × 10³, respectively. In this model, CY-09 administration has no effect on humoral immune response. In the secondary (intranasal) model, BaP significantly increase specific IgE and IgG1 production. CY-09 but not TLR4-IN-C34 administered in combination with BaP significant (p < 0.05) and approximately 2 times enhances specific IgE but not IgG1 production. Specific IgE titers in the control group without inhibitor and in the CY-09 group were (2.0±0.4) × 10² and (5.1±0.3) × 10², respectively.

So, PRR-activation, in our case activation of TLR4 in the model based on subcutaneous immunization or NLRP3 in the model based on intranasal antigen administration with BaP suppressed the production of allergen-specific IgE, but not IgG1. These data are in consistent with the hygiene theory of allergy development.

The goal of research was to determine the frequency of detection and the structure of causally significant sensitization to the main groups of allergens in children with allergic diseases in Siberia when evaluating the results by enzyme immunoassay and immunofluorescence analysis. We researched 968 children with allergic rhinitis, bronchial asthma, urticaria, and/or allergic dermatitis undergoing treatment at the Consultative and Diagnostic Center for Allergology and Immunology, GDKB No. 2 named after V.P. Bisyarina of Omsk. Blood serum samples were diagnosed in the laboratory of clinical immunology. Also, the allergen-specific IgE concentrations were determined for 109 allergens, including food allergens (milk, egg, meat, fish, cereals, fruits, vegetables), household allergens (epidermal pets, dust and house dust mites, molds), and inhaled antigens (grass, trees). Analysis of 435 children was carried out by enzyme immunoassay, and 533 children by indirect immunofluorescence. The research revealed the predominance in children with allergic diseases in Siberia of IgE-mediated sensitization to household allergens, which were detected in 33.7% of the examined. IgE-mediated sensitization to inhalation and food allergens was 1.5 times less common. According to our research, the most common causative allergen among households was house dust (90%), dandruff and animal epithelium (86%), house dust mites (13.5%), and molds (1.4%). The most common causative allergen among pollen was tree allergens (47.8%), cereals (23.5%) and weeds (20.1%). The presence of multivalent sensitization with a predominant combination of household and respiratory allergens and a rarer combination of household and food allergens has been experimentally established.

The important role of epicardial (EAT) and thymic (TAT) adipose tissue in the development of atherosclerosis in patients with coronary artery disease (CAD) is widely discussed. The purpose of the study was to investigate the lymphocyte subsets and FoxP3⁺Treg lymphocytes in epicardial, thymic and subcutaneous adipose tissue depending on the severity of coronary atherosclerosis in patients with chronic CAD. We examined 24 patients with CAD (21 men; mean age 65.0 (58.0-68.0) years) scheduled for open-heart surgery. In samples of EAT, TAT and subcutaneous adipose tissue (SAT), the content of CD4⁺, CD8⁺, B lymphocytes, NK and NKT cells, CD4⁺CD25^hiFoxP3⁺ and CD4⁺CD25^lowFoxP3⁺T regulatory lymphocytes (Treg) and a proportion of Tregs with FoxP3 nuclear translocation was determined by imaging flow cytometry. Depending on the severity of atherosclerosis, assessed according to Gensini Score, patients were divided into groups: group 1 – patients with Gensini Score < 65; group 2 – patients with Gensini Score ≥ 65. Patients in group 2 had higher frequency of EAT CD4⁺CD25^lowTreg with FoxP3nuclear translocation, TAT CD8⁺T lymphocytes and NK cells, a lower content of TAT double positive CD4⁺CD8⁺T lymphocytes, and a tendency towards a decrease of frequency of TAT CD4⁺CD25^hiTreg with FoxP3 nuclear translocation compared to patients in group 1. The level of nuclear translocation of FoxP3 in CD4⁺CD25^hiTreg cells in TAT was inversely related to the proportion of CD8⁺T lymphocytes (r_s = -0.653; p = 0.012) and NK cells (r_s = -0.723; p = 0.003) in TAT, and directly – to the proportion of double positive CD4⁺CD8⁺T lymphocytes in TAT (r_s = 0.567; p = 0.034) and the value of the waist-to-hip ratio (r_s = -0.474; p = 0.041). Further research is required to study the molecular mechanisms of these relationships in patients with coronary atherosclerosis and chronic coronary artery disease.

Currently, statins are the main preparations of anti-atherosclerotic therapy due to a number of effects that reduce the progression of atherosclerosis, including anti-inflammatory effectiveness. The purpose of this study was to evaluate the inflammatory response of monocytes in patients with severe atherosclerosis during therapy with hydrophilic and lipophilic statins, as well as in patients with atherosclerosis not receiving lipid-lowering therapy. A total of 60 patients with severe atherosclerosis of the coronary arteries were included in the study in three groups: 1) receiving atovastatin therapy for at least 12 months before inclusion in the study, n = 20; 2) receiving rosuvastatin therapy for at least 12 months before inclusion in the study, n = 20; and 3) those who had not received statin therapy within a year before inclusion in the study, n = 20. The primary culture of monocytes from study participants was obtained by gradient centrifugation followed by immunomagnetic separation of CD14⁺ monocytes. The isolated cells were cultured for 7 days without stimulation and with pro-inflammatory stimulation using lipopolysaccharide (LPS). The level of basal, LPS-stimulated and re-stimulated secretion of TNFα and IL-1β was determined by enzyme immunoassay. Basal secretion of TNFα and IL-1β in patients receiving statins was lower than in patients who did not receive statins for a year; the secretion of both cytokines was significantly lower in the rosuvastatin group. LPS-stimulated TNFα secretion was significantly lower in the groups of patients receiving statins; IL-1β secretion was significantly lower in the atorvastatin and rosuvastatin groups compared to the group without statins. Re-stimulated IL-1β secretion did not differ significantly between groups; re-stimulated TNFα secretion was significantly lower in the rosuvastatin group compared to the atorvastatin and non-statin groups. Thus, the results of the study demonstrate the anti-inflammatory effectiveness of rosuvastatin, expressed in a decrease in the secretion of pro-inflammatory cytokines by cultured monocytes/macrophages of patients with severe coronary atherosclerosis.

In recent years, the role of sluggish nonspecific inflammation in the pathogenesis of hypertension has been proven. Oxidative stress that develops in hypertension leads to damage of endothelial glycocalyx with impaired barrier and adaptive functions of endothelium. Activation of transcription factor NF-kB leads to increased synthesis of free oxygen radicals, adhesion molecules (VCAM-1, ICAM-1, P-selectin, E-selectin) and proinflammatory cytokines (TNFα, IL-1, IL-6), triggering the inflammatory process in the endothelium. Modern antihypertensive drugs, affecting various pathogenetic mechanisms of arterial hypertension development, do not fully affect the immune component of arterial hypertension. Therefore, the search for various methods affecting the immunologic reactivity of hypertensive patients remains relevant in our time.

The aim of the study was to reveal the effect of interval hypoxytherapy on the state of immunologic reactivity of patients with hypertension stage I.

One hundred seventy male patients of 30-45 years old (mean age 38.36±1.64 years) with hypertension stage I (n = 170) who underwent combined treatment including drug therapy and normobaric interval hypoxic therapy in hypoxia-normoxia mode (oxygen content in hypoxic gas mixture was 20.9%) were examined. Indices of redox status and immunologic reactivity before and after interval hypoxytherapy were studied.

Having a significant antioxidant effect, interval hypoxytherapy led to the subsidence of oxidative stress, which was indicated by an increase in the activity of superoxide dismutase and glutathione peroxidase in blood erythrocytes and a decrease in the content of malonic dialdehyde in blood as a result of a decrease in the generation of free oxygen radicals and suppression of lipid peroxidation processes. Interval hypoxytherapy resulted in a statistically significant increase in the initially decreased content of CD3⁺T lymphocytes (p < 0.05), CD3⁺CD4⁺T lymphocytes (p < 0.02), CD3⁺CD8⁺T lymphocytes (p < 0.02). Initially increased number of B-lymphocytes CD19⁺, CD16⁺ lymphocytes, CD95⁺ lymphocytes significantly decreased. An important result of interval hypoxytherapy was a pronounced anti-inflammatory effect: statistically significant decrease in the content of pro-inflammatory interleukins IL-1β (p < 0.02), IL-6 (p < 0.01) and increase in the content of anti-inflammatory cytokines IL-4 (p < 0.005) and IL-10 (p < 0.005), which led to the subsidence of sluggish nonspecific inflammation.

Suppression of oxidative stress and normalization of immunological reactivity of hypertensive patients after interval hypoxytherapy led to the subsidence of sluggish nonspecific inflammation, reduction of endothelial dysfunction and restoration of the structure and tone of the vascular wall with a decrease in blood pressure and improvement of the clinical course of hypertension.

Environmental and climatic factors have a significant impact on the immune system, shaping its composition and functions. The semiarid region of Aleppo, characterized by hot, dry summers and moderate, wet and cold winters, is also characterized by environmental pollution such as air pollutants and industrial chemicals. The interaction of these environmental and climatic factors can impair immune function by causing inflammatory responses, compromising the integrity of immune cells, and impairing the body’s ability to mount an effective immune response. To date, the physiological characteristics of the parameters of the human blood system and the development of physiological reactions of adaptive immune homeostasis depending on the territory of residence have not been sufficiently studied. In this work, we studied the state of immune indices in women living in the semi-arid region. Thirty women aged from 20 to 60 years without chronic and acute diseases were examined. Immunocompetence was assessed using white blood cell count, lymphocyte subset analysis and calculation of immune indices including neutrophil to lymphocyte ratio (NLR), lymphocyte to monocyte ratio (LMR), systemic inflammatory response index (SIRI), immunoregulatory index (CD4⁺/ CD8⁺) and lymphoproliferation ratio and apoptosis (CD10⁺/CD95⁺ and CD71⁺/CD95⁺). The results of the analysis showed that the median NLR was 2.18 (1.50-2.90), LMR 6.66 (5.47-8.87), SIRI 0.64 (0.45-0.98), immunoregulatory index 0.99 (0.84-1.27) and the ratio of lymphoproliferation processes to apoptosis (CD10⁺/ CD95⁺ and CD71⁺/CD95⁺) 0.95 (0.65-1.29) and 0.94 (0.78-1.11), respectively, which indicates a stressful state of immune homeostasis and signs of premature aging of the immune system. Neutrophil-lymphocyte ratio (NLR), lymphocyte-monocyte ratio (LMR), systemic inflammatory response index (SIRI), immunoregulatory index (CD4⁺/CD8⁺) and lymphoproliferation-apoptosis ratio were determined. (CD10⁺/ CD95⁺ and CD71⁺/ CD95⁺) in the examined women of the semiarid climate zone of residence, emphasizes the stressful level of immune homeostasis. The results obtained highlight the need for systematic individual biomedical monitoring to maintain immune homeostasis in populations inhabiting similar environmental conditions.

Severe immune dysfunction occurs in the majority of cancer patients. One of the methods for overcoming immune disorders is transplantation of peripheral blood hematopoietic stem cells, both autologous and allogeneic, and transfusion of allogeneic natural killer cells (NK cells). Umbilical cord blood is a known source of not only hematopoietic stem cells (HSCs) but also NK cells, characterized by the rapid availability of blood samples from public cord blood banks. Purpose of the study: to analyze the safety of simultaneous transfusion of three allogeneic cord blood to patients with relapsed/refractory multiple myeloma in the early period (7 days after administration), and also to study the response of subpopulations of peripheral blood lymphocytes of patients to this intervention.

The pilot study included 10 patients (7 men and 3 women) with relapsed/refractory multiple myeloma who had previously received more than 3 lines of therapy. The age of the patients was 59.4±3.4 years; all of them received a single transfusion of three allogeneic samples of umbilical cord blood with an average dose of leukocytes of 60.50±8.2 × 10⁸ (NK cells – 4.36±1.2 × 10⁸) per one patient. A total of 30 samples of umbilical cord blood, alloreactive for KIR receptors of the receptor-ligand type, were transfused. The selection of HSCs concentrates was carried out in the umbilical cord blood bank of the Dynasty Medical Center. Before transfusion of umbilical cord blood, patients received cyclophosphamide 5 mg/kg. After transfusion, patients received five subcutaneous injections of IL-2 (roncoleukin) in the amount of 1 million units per day. Observation at the time of assessment of preliminary results: 7 days after transfusion. Analysis of the immunophenotype of patient lymphocytes was studied twice: on the day of transfusion before the start of the protocol and 7 days after transfusion of umbilical cord blood.

No early adverse reactions to transfusions were observed.

Simultaneous transfusion of three allogeneic cord blood to patients with relapsed/refractory multiple myeloma is a safe procedure. There are no adverse reactions, serious adverse events, or significant reactions/changes in the immunophenotype of patient lymphocytes in the early observation period within 7 days after the intervention.

The article presents data on the state of the proteolysis system and genes for the regulation of matrix metalloproteinases in 180 conditionally healthy women and men of young, middle, elderly and senile ages. A single sampling of the material (buccal epithelium) was performed in patients. PCR examination of the buccal epithelium for the analysis of polymorphisms of the genes MMP-1 rs1799750 and MMP-3 rs 3025058 was carried out on the instrument base of the DNA amplifier Bio-Rad CFX96. Serum levels of MMP-1, MMP-3 were studied by the sandwich variant of solid-phase enzyme immunoassay. Statistical processing of the obtained results was carried out using the IBM SPSS Statistics 26 program using nonparametric statistics methods. The results of the study. The values of MMP-1 and MMP-3 in the group of conditionally healthy women at a young age were lower than the values of middle age, whereas in old age their serum levels varied in different directions: MMP-1 decreased, and MMP-3 increased. When analyzing polymorphisms of the MMP-1 rs1799750 gene, the homozygous genotype -/- with low expression of the MMP-1 gene prevailed in young and middle-aged women, and in women over 75 years of age, the carriage of the G/G homozygote with active gene expression was recorded. When evaluating the expression of the MMP-3 rs 3025058 gene, it was found that young women significantly more often had 5A/5A polymorphism with high gene activity, but with a lower level of MMP- 3. In middle and old age, 65% have a heterozygous 5A/6A allele with reduced gene expression activity. In conditionally healthy men, the level of MMP-1 in blood serum increased in the middle-aged and senile groups, the values of MMP-3 in the group under 45 years of age were low compared with the elderly group (p = 0.041). MMP-1 rs1799750 in men under 75 years of age was identified in the -/G variant in the majority. Homozygous polymorphism of the MMP-3 rs 3025058 5A/5A gene with high activity of MMP-3 prevailed in groups of men under 60 years of age with serum levels of MMP-3 100.2 ng/mL and 92.5 ng/mL, respectively.

There is a known autoimmune disease, which is called ‘anti-NMDA receptor encephalitis’, or ‘Dalmau encephalitis’ (named after its discoverer). It is associated with the presence of high titers of antibodies to a certain epitope of the NR1 subunit of NMDA receptors in the blood and/or in the cerebrospinal fluid. This disease in its classic form is accompanied by severe mental (depression, delusions, hallucinations, anxiety, sleep disorders, cognitive impairment, catatonia, etc.) and neurological (convulsions, dyskinesias) symptoms, disturbances of consciousness and autonomic regulation (regulation of respiration, temperature, blood pressure, heart rhythm), as well as with suicidality, which seems to be disproportionate to the severity of mental disturbances. However, there are also ‘non-classical’ forms of anti-NMDA receptor encephalitis. These can manifest with exclusively psychiatric symptoms (for example, depression, psychosis, suicidal behavior), without characteristic neurological symptoms and/or disorders of autonomic regulation. In some of these cases, the correct diagnosis of anti-NMDA receptor encephalitis was delayed by months, years or even decades. In addition, in recent decades, increasing evidence had accumulated that in some patients with so-called ‘classical’ mental disorders, such as schizophrenia, bipolar affective disorder, depressive disorders – and especially in those who show resistance to traditional psychopharmacotherapy – elevated titers of anti-NMDA receptor antibodies can be detected in their blood and/or in their cerebrospinal fluid. It has also been shown that elevated blood titers of anti-NMDA receptor antibodies are often detected in post-mortem examinations of suicide victims. Anti-NMDA receptor antibody titers in psychiatric patients correlate well with a recent history of suicide attempts and/or with current active suicidal ideation. Some of these patients may benefit from aggressive immunosuppressive therapy that is similar to the regimens normally used to treat ‘classical’ anti-NMDA receptor encephalitis with neurological and autonomic manifestations. This might stop their suicidal ideation and/or help in overcoming their resistance to psychopharmacotherapy. This article presents a description and analysis of three clinical cases of patients with various mental disorders, acute suicidality and elevated titers of anti-NMDA receptor antibodies, in whom such therapy has been effective.

The formation of post-stress disorders in veterans of modern military conflicts is due to the peculiarities of the mutual influence of mental and immuno-endocrine processes aimed at maintaining the stability of the body in conditions of chronic activation of physiological systems through a process known as allostasis. The purpose of the study was to study the levels of stress mediators of hypothalamic-pituitary and adrenal origin, the cytokine profile of blood of veterans of modern wars with PTSD.

38 veterans of a special military operation in Ukraine with a diagnosis according to ICD 10: PTSD (F43.1) took part in the study. The diagnosis was verified on the basis of neuropsychological and pathopsychological examination. The comparison group included 30 veterans of the Chechen military campaign of the same age. The levels of stress hormones in the blood were determined by ELISA using the following method: ACTH (IBL, Germany); norepinephrine (Cloud-Clone, China); cortisol (HemaMedica, Russia); dihydroepiandrosterone (DBC, Canada). The blood cytokine profile was determined using a multiplex analysis using the Bio-Plex test system (MERZ, Germany).

Evidence of the accumulation of “allostatic load” during the formation of PTSD in SVO veterans was an increase in the blood concentrations of ACTH, norepinephrine, and cortisol, which are plastic constants of the catatoxic strategy of adaptation to the effects of prolonged combat stress. Allostatic reactions in PTSD included changes in the cytokine profile of the blood in the form of increased levels of pro-inflammatory cytokines (IL- 1β, IL-6, IL-12 TNFα) and decreased anti-inflammatory and regulatory cytokines (IL-4, IL-10, TGF-β, IL-2). This neuroinflammatory status may be associated with the development of psycho-behavioral symptoms of PTSD.

The formation of maladaptive changes with the accumulation of “allostatic load” was clinically expressed in the form of PTSD and was accompanied by changes in the neurocytokine blood profile in the form of increased levels of ACTH, norepinephrine, cortisol, IL-lβ, IL-6, IL-12 TNFα, against the background of a decrease in the concentration of dehydroepiandrosterone, TGF-β, IL-4, IL-10, IL-2, which generally reflects the prevalence of the catatoxic adaptation strategy in combatants.

The purpose of the study is to analyze the indicators of cellular immunity in patients with obesity (OB) and type 2 diabetes mellitus (DM2). The study included 15 patients with DM2 (DM2 group), body mass index (BMI) 37.36±1.12 kg/m², 12 patients with OB (OB group), BMI – 38.12±1.51 kg/m² and 15 practically healthy individuals (comparison group – GС): BMI 23.5±0.15 kg/m². The study was performed on a flow cytometer FC-500 (Beckman Coulter, USA) using double combinations of monoclonal antibodies (Beckman Coulter – Immunotech SAS, France). The percentage indicators of the T cell population were assessed, such as total number of T lymphocytes (CD3⁺), number of T helper cells (CD3⁺CD4⁺), cytotoxic T lymphocytes (CD3⁺CD8⁺), natural killer cells – NK cells (CD3^-CD16⁺CD56⁺), natural killer cells with the properties of T lymphocytes – NKT cells (CD3⁺CD16⁺CD56⁺) and B cell population (CD19⁺) lymphocytes, as well as the relative content of lymphocytes carrying activation markers (CD3⁺HLA-DR⁺, CD3⁺CD25⁺), and apoptosis marker antigen CD45⁺CD95⁺. The relative content of B lymphocytes and T lymphocytes in the peripheral blood of the examined individuals did not have statistically significant differences. In patients in the DM2 and OB groups, there was a tendency (p < 0.1) towards an increase in the relative content of T helper cells and a significant (p < 0.05) decrease in the percentage of T cytotoxic lymphocytes relative to GC. This redistribution of lymphocyte subpopulations led to a significant (p < 0.05) increase in the IRI value (arbitrary units): DM2 – 2.87±0.58; OB – 2.30±0.33 vs GC – 1.62±0.15. The relative content of NK cells and NKT cells in the peripheral blood of the examined individuals did not have a statistically significant difference in the magnitude of the indicators. A statistically significant (p < 0.05) increase in the relative content of T lymphocytes expressing the activation marker HLA-DR (CD3⁺HLA-DR⁺) was found in patients in the DM2 and OB groups relative to GC [(%) DM2 – 7.95±0, 81; OB – 6.54±0.24; GC – 4.01±0.91] and a significant (p < 0.05) increase in the percentage of CD45⁺CD95⁺ lymphocytes in patients with OB and DM2 relative to GC [(%) DM2 – 5.84±0.68; OB – 5.16±0.89; GC – 2.78±0.34]. The results obtained indicate the presence of meta-inflammation in patients with DM2 and OB.

Uric acid (UA) is the end product of purine metabolism and a substance that promotes a chronic inflammatory process. One of the mechanisms of inflammation associated with the UA is the ability of its crystals, mainly monosodium urate, to activate NLRP3 inflammasomes, classifying UA and its salt crystals as damage-related molecular patterns (DAMPs). These crystals also activate the complement system, leading to increase in C3a, C4a, and C5a concentrations and excessive consumption of complement system proteins. It has been known for a long time that UA is able to activate complement, but the relationship between hyperuricemia and complement system functional activity, which can be assessed by complement-mediated hemolysis, remains unclear. In this study, we have made an attempt to estimate the UA concentration that does not lead to spontaneous complement system activation. The study assessed the relationships between the parameters of complement functional activity and some blood biochemical data with UA concentration ([UA]) using correlation analysis. The rate (V_lys) and time of 50% hemolysis (T₅₀) were considered as indicators of complement functional activity, and their relationship was demonstrated using exponential functions y = a*e^[x], which takes the form y = [UA]*e^[C3]. Concentration of C3 is the argument of the function, base of degree is the base of the natural logarithm, and the proportionality coefficient equal to the UA concentration. Correlation analysis showed the inverse dependent between function values and the corresponding values of T₅₀ (r = -0.83, p < 0.0001) in the range of UA concentration exceeding 370 umol/L, which is near to the upper limit of the normal level for women and is within the normal range for men. Thus, the approach to assess the effect of UA on complement activation using the analysis of the complement hemolytic activity is effective to demonstrate the pathogenetic function of UA in the development of the inflammatory process without involving inflammasomes through the direct effects on complement activation processes. The relationships demonstrated suggest that the upper limits of the range of “normal” UA concentrations are arbitrary, and their revision is likely advisable.

Hepatocyte growth factor (HGF), produced by mesenchymal cells, stimulates mitogenesis and angiogenesis in tumor cells. Tumor cells of some solid tumors do not secrete HGF. The aim of the study was to evaluate the prognostic significance of HGF expression in tumor tissue in colorectal cancer (CRC). The study included 50 patients with stage II-III colorectal cancer; they underwent radical surgical treatment, followed by adjuvant chemotherapy according to the FOLFOX/XELOX regimen. In primary tumor samples, quantitative PCR was used to assess the level of HGF expression. Statistical processing of the obtained data was carried out using STATISTICA 13.0, BioStat v.7.1., and Jamovi 1.6.8 software. The study aims to study a new marker. Comparison of characteristics in the case of non-normal distribution was carried out using the nonparametric Mann–Whitney U test. Cox and Kaplan–Meier linear regression tests were used to analyze progression-free survival. When discussing the results, we used our previously obtained data on the level of expression of TGF-β and CXCL8 in the tumor tissue. As a result of the studies, it was found that in 60% of tumor samples HGF was not expressed, but it was significantly higher than in the resection line samples. Analysis of relapse-free survival in patients with CRC according to the level of HGF expression (predicted level by proportional hazards assessment – 0.7) showed that the median survival in groups 1 (HGF expression more than 0.7) and 2 (HGF expression less than 0.7) was 23.3 and 62.9 months, respectively (long rank test p = 0.215). It was shown that the level of HGF mRNA in CRC tumors does not depend on age, stage of the disease, and sensitivity to FOLFOX/XELOX chemotherapy. The expression level is significantly reduced in tumors with a KRAS mutation and increased in those with a BRAF mutation, in poorly differentiated tumors. Using the level of HGF expression in the tumor tissue of patients with non-metastatic CRC before the start of chemotherapy to assess the prognosis of the relapse-free period is only possible in conjunction with the expression of TGF-β, CXCL8 in the tissue and the level of CEA in the blood of these patients.

The term “immunity” was introduced in medicine due to Ilya Mechnikov. However, the concept of infection resistance existed in European culture long before Mechnikov in the form of s.c. “vital force”. According to holistic methodology, the history of immunology can be divided into three unequal periods: 1) vitalistic – from antiquity to the discoveries of Mechnikov and Ehrlich; 2) “Mechnikov time” (1890s – 1916); and 3) a post-Mechnikov period (1916 – present). At the beginning of 18^th century, the existence of “vital force” ideas created the possibility to adapt variolation in Europe, which was based on medieval magical-mystical concepts. Without generalizing of variolation experience, Edward Jenner would hardly have been able to discover vaccination. Scientific ideas of Mechnikov were not a categorical denial of the previous period of immunology. For almost 20 years, he himself had to fend off teleological and vitalistic accusations from the side of those biologists who denied the possibility of phagocytes to act expediently. An analogy helped Mechnikov to confirm his intuition was the phenomenon of intracellular digestion of amoebas or ciliates. Besides, he experimentally proved that phagocytosis was the product of evolution and not a “god gift”. However, in his works, Mechnikov also addressed to the category of “healing forces of the body.” Modern immunology now faces a number of theoretical contradictions and methodological difficulties related to reductionists paradigm: as far as the “split” between the world of “immune objects” (immune cells receptors, antibodies, cytokines, etc.) and “subjectness” of its basic categories “self-nonself”, “immune aggression, “immune defence”, etc., Ilya Mechnikov tried to overcome this contradiction substantiating the phenomenon of phagocytosis as functional system acting in accordance with defense purpose. The construction of “holistic” subject-object models of immunity (such as the symbiosis model of plant immunity) could be a way of overcoming the aforementioned contradictions.