Vol 27, No 2 (2024)
- Year: 2024
- Published: 12.08.2024
- Articles: 41
- URL: https://rusimmun.ru/jour/issue/view/33
Full Issue
SHORT COMMUNICATIONS
Genetic modification of primary human B cells to model the process of B cell development in germinal centers
Abstract
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.
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THP1-based cybrid cells with various mtDNA mutations differ by the ability to form inflammatory response
Abstract
Most age-related human diseases are accompanied by chronic inflammation. Modern research is aimed at studying the principles of the formation of the immune response. The reasons why the local inflammatory reaction cannot be resolved and becomes a sluggish chronic form are still unknown. Immune cells secrete cytokines in response to pathogens. To avoid cell death as a result of high concentrations of cytokines and resulting tissue damage, there is a mechanism of innate immune tolerance. Innate immune tolerance involves a decrease in the secretion of proinflammatory cytokines in response to repeated exposure to a pathogen. It is known that mitochondria play an important role in the formation of the immune response. Consequently, impaired mitochondrial function can lead to impaired immune response. To control the quality of mitochondria in the cell, there is a mechanism – mitophagy. Previously, we have created cybrid lines based on the monocytic cell line THP-1. Cybrids were obtained by fusion of THP-1 cells (mitochondria were removed) with platelets from patients. Each of the cybrid lines had the THP-1 nuclear genome and an individual patient’s mitochondrial genome. In our study, we decided to study the ability of cells carrying different mitochondrial genomes to generate a proinflammatory response, as well as to form tolerance in the future. For this purpose, we chose a model of ecdotoxin tolerance. Thus, we stimulated the cybrid lines twice with lipopolysaccharide and then assessed the secretion of the cytokines TNFα, IL-1β, IL-6, IL-8, and CCL2 using ELISA. The cybrids demonstrated two levels of proinflammatory response: high and low. Moreover, cybrids with a high proinflammatory response either did or did not develop tolerance upon repeated stimulation. In our study, cells that differed from each other only in mitochondrial genome demonstrated three types of reactions upon the induction of immune tolerance to LPS. Future studies will improve our understanding of the mechanisms of mitochondrial involvement in pathological processes. It is likely that studies of deficient mitophagy and the role of certain mtDNA mutations in its development will yield promising results.
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Experimental mouse model of pulmonary fibrosis induced by nebulized LPS administration
Abstract
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.
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Surfactant system of the lungs in antiphospholipid syndrome under the conditions of administration of the immunosuppressive agent FTY-720
Abstract
Antiphospholipid syndrome (APS) is a thrombophilic disease in the pathogenesis of which the leading role belongs to antibodies reacting with antigenic determinants of phospholipids. APS is a systemic autoimmune pathology that involves many organs and systems in the pathological process, including the respiratory system. Given the fact that the lung surfactant is mainly represented by phospholipids, it can be presented that the surfactant is damaged and its functioning is impaired in APS. The results of our experiments showed that in antiphospholipid syndrome, there was a decline in the functional activity of the surfactant and a change in its biochemical composition. The introduction of FTY-720 normalized the parameters of the lung surfactant system, which were changed during the modeling of antiphospholipid syndrome. The work shows the effectiveness of the therapy of this autoimmune pathology with immunosuppressor FTY-720, which is a structural analogue of endogenous sphingosine. The effect of the drug is based on the modulation of sphingosine-1 phosphate receptors of lymphocytes. The main effect of the immunosuppressor FTY-720 as a result of receptor interaction is a decrease in the number of circulating lymphocytes. The research involved 85 white random bred male rats: Group 1 (APS modeling) consisted of 30 rats who were intravenously injected with cardiolipin antigen at a total dose of 0.2-0.4 mg per rat every other day for three weeks; Group 2 (control) consisted of 25 rats who were injected with 0.9% NaCl solution according to the same scheme; and Group 3 including 30 rats in which APS was combined with the administration of the immunosuppressive agent FTY-720 (intraperitoneally 1 mg/kg of animal weight). Three weeks later, there was an operation conducted with the purpose of extraction of the bronchopulmonary complex. After extraction of the bronchopulmonary complex, its triple lavage was carried out with 0.9% NaCl solution. The material of the experimental study was a lavage liquid, in which the biophysical and biochemical parameters of the surfactant were studied. Langmuir–Blodgett method was used to identify static, minimal and maximal surface tension. These figures were used to identify integrative indicator reflecting surfactant characteristics – the Clements stability index. The fractional composition of surfactant phospholipids was determined by thin-layer chromatography. The results of our experiments showed that in antiphospholipid syndrome, there was a decline in the functional activity of the surfactant and a change in its biochemical composition. The introduction of FTY-720 normalized the parameters of the lung surfactant system, which were changed during the modeling of antiphospholipid syndrome.
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Avidin-positive mast cells of the liver when exposed to water-soluble silicon for nine months
Abstract
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.
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Analysis of rat jaw bone tissue elemental composition in a model of periodontitis
Abstract
The elemental composition of jaw bone tissue depends on many external and internal factors affecting the body. The structure of bone tissue directly reflects its ability to restrain inflammation and regenerate after exposure to damaging factors.
The purpose of the study is to study the elemental composition of the rat jaw bone tissue during chronic periodontal inflammatory process. A month later, the individual was removed from the experiment by an overdose of ether to isolate the bone segment under study. Next, the bone tissue sample was studied using atomic emission spectroscopy.
In the studied segment of bone tissue, 67 chemical elements were identified in a quantitative range of up to 10-4 mass percent. The highest mass fraction belongs to the basic elements of bone tissue: calcium, phosphorus, magnesium, sodium, and sulfur. While during the experiment the animals received standard complete feed, which contained proteins, vitamin D close to normal, Ca, P and Se. In the bone tissue sample under study, the quantitative Ca/P ratio was 1.685, which slightly exceeds the optimal one. An increase in this indicator is associated with a decrease in phosphorus content in bone tissue, which can be explained by insufficient daily intake of this element from food, as well as a number of local factors. When the jaw is traumatized during the modeling of periodontitis, an inflammatory process occurs in this area, the experimental animal experiences stress, which leads to demineralization of the tissues of the alveolar process of the jaw, as evidenced by a decrease in the content of phosphorus and selenium in the bone tissue. In addition, elemental analysis showed the presence of foreign elements in the bone tissue, the amount of which corresponds to the range from 0.001% to 0.06%. Basically, these are elements that form simple substances - metals: bismuth, gallium, lead, iridium, titanium, zinc, mercury, molybdenum.
The results of the study can be taken into account when planning and conducting polyetiotropic therapy for inflammatory-dystrophic periodontal diseases in the early stages of their development.
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Effect of proteins from the coelomic fluid of superficially wounded holothurians on the activity of phagocytes
Abstract
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.
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On the issue of experimental modeling of invasions and prospects for the use of immunostimulants in nematodes
Abstract
Currently, the search for ways to overcome drug resistance to anthelmintics is relevant. This problem is solved by limiting the duration of use of drugs with the same mechanism of action and combination of anthelmintics. The studies presented in the article have shown that the use of immunotropic drugs is also very promising.
Forty white mice and 20 white rats were used for experimental modeling of Trichinella spiralis and Trichinella nativa invasions. Changes in lymphoid tissue associated in the intestine, in the spleen, as well as in the peripheral blood of animals and birds during inoculation of the above species were studied. Further, the effect of drugs with immunotropic activity was studied on proven models of Trichinella spiralis invasions, for which 30 white mice (20.5±2.5 g) were used, which were randomized and divided into groups. The evaluation of the protective properties of immunotropic drugs in trichinosis was carried out on laboratory models to determine the survival rate in studies after euthanasia of all mice and counting the number of Trichinella larvae. Both experiments were conducted in accordance with the principles of the Helsinki Declaration. After removing laboratory animals from the experiment (in accordance with the principles of the above declaration), macro- and microanatomic studies and sampling of histological material (sets of intestines and spleens) were carried out for qualitative and quantitative assessment.
The above is confirmed by the fact that parasitological studies have established the high effectiveness of polyoxidonium in relation to the survival of Trichinella larvae, which was accompanied by an increase in the area of lymphoid tissue associated with the intestine and the white pulp of the spleen, in which lymphoid follicles grew. Thus, polyoxidonium is an effective drug that can be used in the complex treatment of nematodes as an immunotropic agent.
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Studying the anti-inflammatory activity of Bifidobacterium bifidum supernatants and chicken embryo cells on a model of opisthochic invasion
Abstract
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.
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In situ and in silico modeling of the hematopoiesis-inducing effect of chelidonic acid
Abstract
The current trend in regenerative medicine, in the context of an aging population, is the search for new ways and means to optimize tissue bioengineering. One of the convenient models for in situ studying bone marrow regeneration is the subcutaneous ectopic osteogenesis test on scaffolds that imitate the architecture of bone tissue. Chelidonic acid (CA), a small molecule, is capable of participating in various cellular processes and metabolic pathways, and it can activate the osteogenic differentiation of mesenchymal stem cells. However, the molecular mechanisms behind the regulatory effects of CA remain unknown. The aim of this study was to investigate the modulatory effect of CA on the in situ formation of hematopoietic foci, as well as to predict target genes and intracellular signalling pathways involved in the hematopoietic activity of CA. An aqueous solution of CA, isolated from an extract of the Saussurea controversa plant. Course (daily for 35 days) oral administration of CA. Ectopic osteogenesis testing in Balb/c mice. Morphometric analysis of histological sections after 45 days and in silico modelling of gene expression with statistical analysis. CA, when administered orally in a low dose (10 mg/kg), threefold increases the normalized area of bone marrow in the composition of bone tissue plates grown in situ in a test of ectopic subcutaneous osteogenesis in mice. This effect is associated essentially (a probability of CA activity Pa > 0.5 and a probability of inactivity Pi < 0.5) with enhanced expression of 358 hematopoiesis-related genes, as predicted by in silico analysis. The top list with the highest Pa value included 10 target genes, such as GATA1, CITED2, SFRP1, EP300, LGALS9, VNN1, IL10RB, RARA, CD83, and HMOX1. CA has a significant ability to enhance the reparative remodelling of hematopoietic tissue in situ. The next phase of research will be to test actual target genes and signalling pathways that mediate the regulatory effect of HC on hematopoiesis both in vitro and in vivo, as well as in clinical settings.
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Influence of endomorphins-1, 2 on glucose uptake by T and B lymphocytes in vivo
Abstract
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.
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Mechanisms of RORα-dependent effects of melatonin
Abstract
The transcription factor RORα has not traditionally been attributed a fundamental role in the development of Th17 cells, but recent studies have shown that it is necessary for the formation of a pathogenic variant of Th17 cells, the so-called Th1-polarized Th17 (Th17.1). Since the transcriptional activity of RORα depends on ligand binding, the search for such ligands is highly relevant, and in this regard, melatonin is of particular interest. The question of the ability of RORα to directly bind melatonin remains open today; data on this problem are extremely contradictory. In 1995, I. Wiesenberg and colleagues identified RORα as a nuclear receptor for melatonin, demonstrating the hormone’s ability to enhance the binding of this factor to DNA and determining the dissociation constant for the interaction of RORα with melatonin using classical Scatchard analysis. In 2011, P.J. Lardone and colleagues “rediscovered” RORα as a receptor for melatonin by demonstrating the coprecipitation of melatonin with RORα. And in 2016, A.J. Slominski and colleagues published a paper that cast doubt on the possibility of melatonin binding to RORα based on molecular modeling of ligand-receptor interactions supported by functional studies. However, a careful analysis of these data indicates the ambiguity of this conclusion, allowing us to speak, rather, of medium or low binding affinity of the hormone to RORα, but not of its absence. This conclusion is also supported by the fact that RORα mediates many of the effects of melatonin, both physiological and pharmacological, including the regulation of circadian rhythms and oxidative metabolism, neuro- and cardioprotection, and control of the immune response. In general, the data available today allow us to consider the transcription factor RORα as a receptor for melatonin with medium affinity, although indirect regulation of this factor by the hormone is not excluded, and RORα-dependent mechanisms should contribute to the cellular response to melatonin, both under physiological conditions and in the case of pharmacological use of the hormone.
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Features of mitochondrial state in CD4+Т lymphocyte subsets
Abstract
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.
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Markers of systemic leukocyte activation for assessing acute and chronic systemic inflammation
Abstract
The phenomenon of systemic leukocyte activation is a component of systemic inflammation (SI), known as the systemic inflammatory response (SIR). Currently, there are hundreds of molecular markers that can serve as criteria for systemic activation, including soluble receptor to IL-2 (sIL-2R) or CD25, β2-microglobulin, and eosinophil cationic protein (ECP). The objective of our study was to assess whether markers of systemic leukocyte activation can be used as criteria for acute and chronic systemic inflammation. Data from 121 patients with acute critical infectious conditions – intensive care patients with sepsis-2 (according to the 2001 consensus, without MODS) and sepsis-3 (2016 consensus, with MODS) – and 136 patients with non-infectious genesis (polytrauma with and without MODS) were analyzed to study acute SI. Additionally, data from 148 patients with immune-inflammatory rheumatic diseases were analyzed to study chronic SI. The reactivity level (RL) was calculated using the author’s method to assess the intensity of SIR. The development of SI was verified based on the measured concentrations of IL-6, IL-8, IL-10, TNFα, CRP, D-dimers, cortisol, myoglobin, and troponin I. Criteria for leukocyte activation phenomenon were sIL-2R > 700 units/mL, β2-microglobulin > 3000 ng/mL, and ECP > 10 ng/mL.
Based on the study results, TNFα, ECP, and β2-microglobulin were identified as the most stable indices for dividing groups with and without systemic vasculitis in both acute and chronic diseases. These indices indicate the typical character of systemic leukocyte activation and can be used to assess the intensity of the described phenomenon when integrated into the SI score. However, it is doubtful that these indices can be independently applied as criteria for the complex process of SI and even the SIR phenomenon. This is because the registration of threshold values for two criteria simultaneously during the acute process of SI was only observed in 40.2% of cases, and in chronic processes, it was only observed in 23.7%. These values were comparable to the group without acute SI. Additionally, all three additional criteria were only determined in acute SI in 20.5% of cases and in chronic SI in 3.4%.
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Inhibition of TLR4 and NLRP3 leads to the exacerbation of IgE specific antibodies in mouse allergic models based on subcutaneous or intranasal immunization respectively
Abstract
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.
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Allergy in HIV-Infected patients (on the example of clinical observation)
Abstract
The article is devoted to the study of the causes of the formation of allergic diseases in patients with HIV infection on the example of a clinical case. Patient C., 35 years old, was under medical supervision at the Victory Clinic from 2013 to 2015 with manifestations of skin allergies. Over the next four years, the patient received therapy for exacerbation of atopic dermatitis, followed a diet with the exception of dairy products and beef. The patient’s condition could be assessed as satisfactory; relapses of atopic dermatitis were noted 2 times after a violation of the diet. In 2017, after an unprotected contact, the patient noticed: weakness, weight loss, dry skin and enlarged peripheral lymph nodes. In June 2017, the patient turned to the district pediatrician, who prescribed a referral for a general blood and urine test, a blood test for AIDS and hepatitis. As a result of the examination, the patient was diagnosed with HIV infection. The patient was examined and treated at the Yakutsk AIDS center. Since August 2017, the patient has been worried about nasal congestion, sneezing, watery eyes, and headaches. In early September 2017, the patient turned to an allergist-immunologist. An examination was conducted: a blood test was taken for allergoscreen panel No. 1 and an immunogram, rhinocytogram. The following results were obtained: according to allergoscreen No. 1, allergy to birch 3.0, milk 3.2, wheat flour 2.8. Rhinocytogram data from 09/28/2017: neutrophils 67 per-field and eosinophils 10 per-field. People infected with the human immunodeficiency virus (HIV) have high levels of allergic conditions, including allergic rhinitis (hay fever), drug allergies and asthma. The HIV virus infects and destroys CD4+T cells, a type of white blood cell. This leads to a change in immune function, which contributes to the development of allergies, infections, cancer and other immune problems. In a patient with HIV infection, after the pathology is detected, the formation and transformation of allergic diseases is observed. The formation of allergopathology in HIV patients is associated with a reduced level of CD4+ cells, which is one of the factors contributing to the development of allergy transformation. The treatment of allergic diseases such as: bronchial asthma, atopic dermatitis, allergic rhinitis and allergic urticaria in patients with HIV is the same as in patients not infected with HIV. Oral administration of glucocorticosteroids should be avoided due to the immunosuppressive effects of this group of drugs.
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Identification of IgE-mediated sensitization in children by enzyme immunoassay and immunofluorescence methods
Abstract
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.
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Soluble receptor for advanced glycation endproducts (sRAGE) as a new biomarker of asthma in children: a brief review of the literature and our own findings
Abstract
In recent years, the receptor of glycosylation end products (RAGE) has increasingly attracted the attention of researchers and plays an important role in various in various diseases associated with tissue destruction, metabolic and neurodegenerative processes, infections, immune reactions and inflammation of various origins. The study of the soluble form of this receptor (sRAGE) as a potential biomarker for the diagnosis and monitoring of bronchial asthma (BA) in children is becoming particularly relevant.
This article presents a brief review of the literature on the role of RAGE and its ligands in the pathogenesis of respiratory diseases, and analyzes our own data on the study of sRAGE levels in children with AD.
We examined 101 children aged 7-18 years suffering from AD referred to the Children’s Allergy Center (Krasnoyarsk, Russia). Children with AD were divided into three groups: controlled (n = 45), partially controlled (n = 19) and uncontrolled asthma (n = 37) according to GINA-2023. The control group consisted of 92 age- and sex-matched children (virtually healthy children without signs of infection or allergy). Serum sRAGE levels were determined by magnetic immunoassay (MAGPIX, Luminex, USA; Merk Millipore, USA). Data are presented as median (25-75% quartiles). The Kruskal–Wallis test was used.
Serum sRAGE levels were reduced only in uncontrolled AD in the examined children (Figure 1, p (Kruskal–Wallis test) = 0.001). In addition, serum sRAGE levels were reduced in the asthma exacerbation groups, regardless of exacerbation status caused by viral infection or allergens.
Thus, serum sRAGE levels are closely related to asthma control and exacerbation status in children with AD in and may be used as a novel marker of loss of disease control and possibly as a potential target for therapeutic intervention.
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Lymphocyte subsets in epicardial, thymic and subcutaneous adipose tissue during advanced coronary atherosclerosis in patients with coronary artery disease
Abstract
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+CD25hiFoxP3+ and CD4+CD25lowFoxP3+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+CD25lowTreg 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+CD25hiTreg with FoxP3 nuclear translocation compared to patients in group 1. The level of nuclear translocation of FoxP3 in CD4+CD25hiTreg cells in TAT was inversely related to the proportion of CD8+T lymphocytes (rs = -0.653; p = 0.012) and NK cells (rs = -0.723; p = 0.003) in TAT, and directly – to the proportion of double positive CD4+CD8+T lymphocytes in TAT (rs = 0.567; p = 0.034) and the value of the waist-to-hip ratio (rs = -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.
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Proatherogenic proteomic profile of LDL isolated from plasma of patients with diabetes mellitus: immunological aspects
Abstract
Atherosclerosis is a vascular disease, which is based on a chronic inflammatory process, including complex interactions between blood components, as well as the cells that form the vessel wall, and lipid metabolism in general. Low-density lipoproteins (LDL) play a key role in pathogenesis of atherosclerosis. When the permeability of the endothelial layer of the vessel is impaired, LDL can penetrate into the intramural space and lead to excessive cholesterol accumulation by the cells from the intima subendothelial layer, leading to the onset of the inflammatory process and to the formation of foam cells, the main morphological component of the atherosclerotic plaque. However, the level of LDL in the patient’s blood plasma alone is not a key indicator of the development of atherosclerosis. Numerous studies point out the role of oxidized modifications of LDL (oxLDL) in the increased local accumulation of cholesterol in vascular wall cells, however, recent works have shown conflicting results regarding the role of oxLDL in the development of atherosclerosis. We hypothesize that other components of LDL may also influence the progression of atherosclerosis. It is a well-known fact that patients with diabetes mellitus (DM) suffer from cardiovascular diseases, in particular atherosclerosis, more often than patients without diagnosed DM and other autoimmune diseases, and the disease progresses faster. The purpose of this study was to identify risk biomarkers in LDL groups that indicate the relationship of the immune system with the development of atherosclerosis in such patients. LDL was isolated from patients and healthy donors using continuous ultracentrifugation with solutions of different densities, and LDL protein profile samples were measured using gas chromatography-mass spectrometry. We found 9 proteins that had a statistically significant difference between the samples (DM and control). In the diabetes group, the content of the antimicrobial peptide cathelicidin and lipopolysaccharide-binding protein was almost 2 times higher compared to the control. These proteins may be involved in the development of inflammation, leading to the progression of atherosclerosis. At the same time, a decrease in immunoglobulins and complement components (C9 and Complement C1s subcomponent) associated with LDL may contribute to the development of atherosclerosis.
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Effect of statin administration on the inflammatory response of monocytes in patients with atherosclerosis
Abstract
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.
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Dynamic features of the humoral immunity in patients with acute coronary syndrome depending on the content of B lymphocytes with the CD3-CD19+CD5+ phenotype, who have and have not COVID-19
Abstract
The purpose of the work was to study the dynamic features of the humoral immunity in patients with acute coronary syndrome depending on the content of B lymphocytes with the CD3-CD19+CD5+ phenotype, who had and did not have COVID-19. We analyzed data on humoral immunity in men aged 40 to 65 years with acute coronary syndrome, who had and did not have COVID-19, depending on the content of B lymphocytes with the CD3-CD19+CD5+ phenotype. All patients underwent coronary angiography with further stenting of the coronary arteries. The values of lymphocytes with the CD45+CD3-CD19+ phenotype and the presence or absence of COVID-19 previously were taken into account. Using the flow cytometry method, 2 groups were gated using the pan-leukocyte marker CD45+: the subgroup of B lymphocytes CD3-CD19+CD5+ and the subgroup of B lymphocytes CD3-CD19+CD5-, B lymphocytes (CD45+CD3-CD19+). Levels of total IgA, IgG, IgM, C1-inhibitor, C3a and C5a complement components by enzyme immunoassay.
After coronary artery stenting, B lymphocytes (CD3-CD19+CD5+) significantly increased in the group of people with COVID-19, with initially low levels of these cells (p < 0.01) and normal cells (p < 0.05). In individuals who had COVID-19 and did not have this disease, with initially decreased CD3-CD19+CD5+ cells, B lymphocytes (CD45) increased significantly (p < 0.001 and p < 0.05, respectively) over time (p < 0.05+CD3-CD19+), B lymphocytes (CD3-CD19+CD5-). In the group of patients who recovered from COVID-19 with normal CD3-CD19+CD5+ cells, the relative number of B-lymphocytes decreased over time (p < 0.01) (CD45+CD3-CD19+) and CD3-CD19+CD5- cells. In persons who had previously suffered from COVID-19, with increased CD3-CD19+CD5+ cells, the number of B-lymphocytes (CD45+) significantly (p < 0.05) decreased over time CD3-CD19+), in those who did not have COVID-19 with high above-mentioned cells, immunoglobulin A significantly (p < 0.05) increased. The most severe clinically was the group of people with low CD3-CD19+CD5+B cells and COVID-19 in the anamnesis.
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Effect of interval hypoxytherapy on the immune status of hypertensive patients
Abstract
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.
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Interrelations between iron content and immune dysfunctions in frequently ill children of the Aral Sea Region
Abstract
The widespread occurrence of immune-dependent diseases and disturbances in iron metabolism, as well as their co-occurrence and the impact of iron deficiency on the immune system, underscore the relevance of this study. Modern data also indicate the involvement of iron in regulating cytokine production and protein synthesis by lymphocytes. In addition to the age-related characteristics of the child’s immune system, a significant factor in the development of inadequate course of the infectious process is the disruption of immune system adaptation, which can be caused by frequent infections, other somatic diseases such as anemia, and environmental factors. The aim of this study is to assess the relationship between iron levels and immune dysfunction in children with frequent acute respiratory diseases living in the Aral Sea region. A total of 127 children, who are native residents living at the epicenter of the ecological crisis, were examined. The children’s ages ranged from 1 to 5 years old. Among them, there were 105 frequently ill children (with 6 or more episodes of acute respiratory infections per year) with iron-deficiency anemia (IDA), and 22 healthy children of the same age. All children underwent clinical examination and standard laboratory tests, including complete blood count, relative content and ratio of T and B lymphocytes, as well as the levels of pro- and anti-inflammatory cytokines (IL-2 and 4) and ferritin in the serum.
According to the results of the conducted research, it was found that among frequently ill children with iron-deficiency anemia (IDA), there is a moderate positive correlation between the relative quantity of T lymphocytes and the hemoglobin level (r = +0.62), as well as a strong positive correlation with the serum iron level (r = +0.78). This indicates a significant influence of iron deficiency on the level of cellular immunity. Clinical data also indicate a higher risk of respiratory infections development in children with IDA compared to the control group. Ferritin, hemoglobin, lymphocytes, and cytokines act as positive regulators of hematopoiesis in IDA conditions and contribute to the compensation of anemia during iron deficiency replenishment.
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Ratio of immune indices in women of the semi-arid region
Abstract
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.
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Immunotherapy is a new opportunity for managing patients with “thin” endometrium in cycles of transfer of thawed embryos
Abstract
The trend of recent decades has been the search for therapeutic effects on the so-called “thin” endometrium. However, to date there is no optimal management scheme for patients with “thin” endometrium in assisted reproductive technology programs. Aim: to evaluate the effectiveness of using recombinant IL-2 before cryotransfer in women of late reproductive age with “thin” endometrium and repeated implantation failure after transfer of good quality embryos.
The study included 63 patients of late reproductive age with “thin” endometrium. The first group consisted of 25 patients of late reproductive age with a history of unsuccessful in vitro fertilization programs with good quality embryos. Cryotransfer to the patients of the first group was carried out in a modified natural cycle. The second group included 38 women of late reproductive age with a history of unsuccessful in vitro fertilization programs with good quality embryos. Cryotransfer for women of the second group was carried out in a cycle with hormone replacement therapy. Recombinant IL-2 in the cycle before cryotransfer were prescribed to all women. In both groups, implantation, clinical pregnancy, and live birth rates following administration of recombinant IL-2 were valuable. In both groups, the implantation rate, clinical pregnancy rate and live birth rate were assessed after administration of recombinant IL-2. Statistical data processing was performed using application software package Statistica 6.0 (StatSoft, Ink. 1994-2001), adapted for biomedical research. p < 0.05 was considered significant.
After recombinant IL-2 treatment of patients with infertility and thin endometrium, endometrial thickness increases statistically significantly in the first two menstrual cycles after treatment, so transfer of thawed embryos should be planned in the first two menstrual cycles after immunotherapy.
In order to increase the thickness of the endometrium during cryotransfer in a modified natural cycle, as well as in a cycle with replacement hormone therapy, recombinant IL-2 can be used in women with “thin” endometrium with repeated unsuccessful implantation attempts when transferring good quality embryos.
Cryotransfer should be performed in the first two menstrual cycles after immunotherapy.
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Transfusion of allogenic cord blood to patients with multiple myeloma: safety analysis and influence on the immunophenotype of blood lymphocytes in the early period
Abstract
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.
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Pleiotropicity of immunomodulating effects of synthetic thymic hexapeptide in chronic infectious and inflammatory diseases of the genital tract in immunocompromized women
Abstract
A reason for a protracted course and frequent recurrence of pelvic inflammatory diseases (PID) is dysfunction of the immune system, which dictates the need of effective immunotherapeutic approaches. Objective: to clarify the features of immune system functioning, the interferon system, the profile of cytokines during the period of exacerbation of PID in women; to develop targeted immunotherapy using hexapeptide with assessment of its effectiveness. The studied immunocompromised women 20-40 years old with exacerbation of PID were divided into study groups (SG): SG1 (n = 22), SG2 (n = 70) before treatment; SG1a, SG1 receiving standard therapy; SG2a, SG2, receiving standard therapy and hexapeptide (HP, Imunofan®). The parameters of cellular and humoral immunity, levels of serum interferons, cytokines were determined. During exacerbation of PID, a decrease in IFNα was in both groups and IFNγ, more pronounced in SG1. In SG1, the levels of IL-6, TNFα were increased, the level of IL-8 was decreased. In SG2, the level of IL-6 was increased. An increase in IL-4, IL-10 was noted in both groups. Against the background of IFNα, IFNγ deficiency and cytokine imbalance, a significant decrease in T lymphocytes, T helper cells, B lymphocytes was observed in both study groups. After standard treatment, there was no recovery of IFNα, IFNγ. Only TNFα decreased, the levels of IL-1β, IL-6 were increased at the end of treatment, there was no increase in IL-4, IL-10 and restoration of cellular and humoral immunity. It leads to maintaining chronic inflammation and exacerbation of PID within 3 months after treatment. After immunotherapy including fHP, IFNα was restored in 50% cases. There was a decrease in IL-1β, TNFα without significant changes in the levels of IL-8, IL-18. Resolution of inflammation was accompanied by an increase in IL-10. The modulating effect of fHP on interferons and cytokines contributed to the restoration of T cell and humoral immunity, which is related to the onset of clinical remission of PID at an earlier time and an increase in the duration of the inter-relapse period. The positive effectiveness of immunotherapy with Imunofan® is due to its pleiotropic effects, which provides significant advantages compared to the use of standard therapy only.
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Level of some matrix metalloproteinases in blood serum in relation to their genetic polymorphism depending on age
Abstract
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.
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Frequency of polymorphism of the C-589T IL4 and G-1082A IL10 genes in children with new coronavirus infection
Abstract
Immune responses are involved in the initiation and development of COVID-19. Compared with mild COVID-19 cases, serum interleukins levels were significantly increased in severe and critical COVID-19 patients. Increases in IL-4 and IL-10 are associated with disease severity. The C-589T polymorphism of IL4 gene is associated with increased promoter activity of gene. Allelic variants GA and GG-1082 of IL10 gene have greater activity compared to the AA-1082 IL10. The purpose was to study the frequency of occurrence of polymorphic variants of the C-589T IL4 and G-1082A IL10 genes in children with a new coronavirus infection in the population of Crimea. Methods. Examined: 68 patients aged 0 to 14 years with a diagnosis of moderate and severe COVID-19, 100 adult patients (45.6±6.14 years). The frequency of occurrence of polymorphisms of the C-589T IL4 and G-1082A IL10 genes was analyzed. Results. The CC and CT variants of the C-589T IL4 in the group of children were more common than the TT C-589T IL4 variants. Statistically significant differences in the frequency of occurrence of the mutant CC genotype of the C-589T IL4 were shown between groups of children and adults. Statistical analysis did not reveal any genotype differences with disease in either children or adults. Analysis of the prevalence of G-1082A IL10 genotypes showed that in children the most common is GA. The same trend is typical for adults and control group. Conclusions. Analysis of the odds ratio showed that there was no pathogenetic significance between the polymorphism of the C-589T IL4 and G-1082A IL10 genes and the risk of developing a new coronavirus infection in the studied groups.
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Autoimmune mediated suicidal behavior
Abstract
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.
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Central effects of ex vivo caffeine-modulated immune cells in the mechanisms of editing depressive-like behavior
Abstract
Depression is a serious medical and social problem due to its high prevalence, involvement of people of working age and lack of highly effective therapy. Social stressors contribute to the prevalence of depression. The COVID-19 pandemic and the associated rules of social distancing, military clashes, and a deteriorating economic situation can lead to a painful “breakdown” of socio-biological adaptation mechanisms and contribute to an increase in the prevalence of depressive disorders, which, according to WHO forecasts, may take second place in the world by 2030, which leads to interest in studying this problem and finding new effective approaches to therapy. Decreased cognitive function in depressive disorders is caused by neuroinflammatory and neurodegenerative changes. The latter are predominantly recorded in the hippocampus, numerous changes in the plasticity of which have been observed both in patients with clinical depression and in rodent models of depression. There is also a sufficient amount of data on the significant role of immune cells and their cytokines in depression, including in the development of behavioral phenotype. We have previously shown that spleen cells of depressive-like mice after ex vivo treatment with caffeine, a psychoactive drug with a wide range of immunomodulatory properties, change their functional activity and, after intravenous administration to syngeneic depressive-like recipients, have editing depressive-like behavior effect. The purpose of this work was to investigate the central effects of caffeine-modulated spleen cells in the mechanisms of editing depressive-like behavior. It was found that in depressive-like recipients after transplantation of syngeneic caffeine-modulated splenocytes, there is an increase in the density of neurons in the CA1 and CA3 zones of the hippocampus, accompanied by BDNF level increase in the hippocampus and prefrontal cortex against the background of a decrease of a number of pro-inflammatory (IL-1β, IL-6, IFNγ and TNFα) and increased of anti-inflammatory (IL-10 and IL-4) cytokines in brain structures pathogenetically significant for the state of depression. The mechanisms of the identified structural and functional changes in the recipient’s brain after the caffeine-modulated splenocytes transplantation are discussed, including their possible direct influence, confirmed by visualization of transplanted cells in the brain parenchyma of depressed-like recipients.
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Neurocytokine blood profile of veterans of modern combat conflicts with post-traumatic stress disorder
Abstract
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.
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Distribution of the T cell element of the immune system in post-COVID patients with first identified hyperglycemia
Abstract
Post-COVID syndrome combines many adverse outcomes with often new-onset conditions, including type 2 diabetes mellitus (DM). The relevance of the study is the insufficient data on T cell immunity in patients with hyperglycemia after COVID-19. The purpose of this study was to identify the characteristics of T cell immunity in individuals with post-COVID syndrome and newly diagnosed carbohydrate metabolism disorders (CDM) in the post-COVID period. The study included 100 patients (32 men and 68 women) aged from 38 to 70 years (mean age (M±m) was 59.40±8.80 years) with a verified post-COVID-19 condition. All patients were divided into three groups: group 1 – patients with concomitant newly diagnosed impaired glucose tolerance (IGT) in the post-COVID period (prior to SARS-CoV-2 infection, no increase in glycemia was detected according to primary medical documentation) (n = 30), group 2 – patients with concomitant newly diagnosed type 2 diabetes mellitus (n = 30), and group 3 – patients with the development of post-COVID syndrome without hyperglycemia (n = 40). During the study, the following parameters were determined: CD45+, CD3+ (T lymphocytes), CD45+, CD3+, CD4+ (helper inducers), CD45+, CD3+, CD8+ (cytotoxic T lymphocytes), CD45+, CD3+, CD4+, CD25+, CD127- (T regulatory cells), CD45+, CD3+, CD4+, CD25+ (T lymphocytes – early activation), CD45+, CD3+, HLA-DR+ (T lymphocytes – late activation). Infection with SARS-CoV-2 with the subsequent development of post-COVID syndrome and persistent hyperglycemia (IGT, type 2 diabetes) in the post-COVID period was accompanied by a higher level of the total number of T lymphocytes. A study of the T lymphocyte population revealed changes in the subpopulation composition. In the group of patients with CDM, a significant increase in T cytotoxic and T regulatory cells was noted. More pronounced changes in the subpopulation composition were identified in the group of patients with newly diagnosed type 2 diabetes in the post-COVID period: an increase in the relative and absolute numbers of T helper cells and T cells of late activation of lymphocytes. Disorders of carbohydrate metabolism associated with COVID-19 are accompanied by changes in the subpopulation composition of T lymphocytes. In individuals with newly diagnosed type 2 diabetes in the post-COVID period, hyperactivation of the T cell component of the immune system was revealed. Heterogeneous data were obtained in individuals with newly diagnosed impaired glucose tolerance, which requires further study of this group of patients.
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Activity of cellular immunity in patients with nutrition-dependent diseases (obesity, type 2 diabetes)
Abstract
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.
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Systemic imbalance of TGF-β isoforms in patients with various manifestations of diabetic retinopathy
Abstract
Transforming growth factor-β (TGF-β) is a group of structurally similar multifunctional regulatory proteins that exhibit pleiotropy in various tissues of the body. Considering that TGF-β isoforms show tropism to various cells, and in diabetic retinopathy both a neurodegenerative process and damage to microvessels can be simultaneously realized, we believe that the study of the content of TGF-β1, TGF-β2, and TGF-β3 in patients with various manifestations of diabetic retinopathy (DR) is relevant. The aim of the study was to determine the levels of TGF-β1, TGF-β2, and TGF-β3 in the blood serum in patients with neurodegenerative and vascular manifestations of diabetic retinopathy against the background of type 2 diabetes mellitus. The study involved 80 patients diagnosed with type 2 diabetes mellitus and 30 healthy volunteers. Based on ophthalmoscopy and optical coherence tomography (OCT), patients with type 2 diabetes mellitus were divided into 4 groups: Group 1 (n = 12) – patients without vascular symptoms of fundus DR and without OCT signs of retinal neurodegeneration; Group 2 (n = 28) – patients without vascular symptoms of fundus DR and the presence of OCT signs of retinal neurodegeneration; Group 3 (n = 10) – patients with vascular symptoms of non-proliferative DR and without OCT signs of retinal neurodegeneration; and Group 4 (n = 30) – patients with vascular symptoms of non-proliferative DR and the presence of OCT signs of retinal neurodegeneration. The serum content of TGF-β1 in all groups was within the reference values throughout the study. The serum concentration of TGF-β2 in all groups was higher in comparison with the control group. The content of TGF-β3 in the blood serum of patients with neurodegenerative manifestations of diabetic retinopathy (groups 2 and 4) was significantly lower than the reference values. A more pronounced deficiency was observed in group 4, whose patients had both vascular and neurodegenerative signs of DR. The presented study showed the presence of a systemic imbalance in the levels of TGF-β1, TGF-β2, and TGF-β3 in vascular and neuronal manifestations of diabetic retinopathy. The results obtained confirm the data that changes in the production of cytokines from the TGF-β family in one pathological process can be multidirectional.
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Relationship between serum uric acid levels and complement functional activity
Abstract
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 (Vlys) 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.
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Relationship of the phenotype of peripheral blood lymphocytes and signs of osteopenia in patients with chronic lymphocytic leukemia
Abstract
Chronic lymphocytic leukemia (CLL) is the most common leukemia among adults in Western countries, characterized by the development of a number of complications, including osteoporosis, which is a prerequisite for studying its predictors. The purpose of the work is to investigate the relationship between immunophenotyping indicators of blood lymphocytes and osteodensitometry indicators in CLL. The study was conducted on 48 male patients with CLL aged 50-70 years with an average disease duration of 12 months and 14 apparently healthy men of the corresponding age (group 1). In the blood, CD5+, CD19+, CD20+, CD22+, CD23+, CD43+, and CD200+ lymphocytes were differentiated in the blood using flow cytometry. The international FRAX questionnaire was used to assess the probability of fracture. Osteodensitometry assessed bone mineral density (BMD), T-score (standard deviation, SD), Z-score (SD) in the lumbar spine (LS), proximal femoral neck (FNA), proximal femoral bone (PFB). Patients with CLL were included in group 2 (n = 34) with BMD within normal limits (T- and Z-score > – 1.0 SD) or group 3 (n = 14) with signs of osteopenia (OP) (T- and Z-score from – 1.0 SD up to – 2.5 SD). In patients with CLL in group 3, compared with patients with CLL in group 2, there is a significant increase in the number of lymphocytes in the blood expressing markers CD5+, CD19+, CD20+, CD22+, CD23+, CD43+, and CD200+. In patients with CLL in group 3, signs of AP in the SPBC increase with the increase in lymphocytes with the CD5+, CD19+, CD20+, CD22+, CD23+, CD43+, CD200+ phenotype in the blood. Signs of AP in the PBC increase with the increase in lymphocytes with the CD5+, CD19+, CD20+, CD23+, CD43+ phenotype in the blood. The strongest connections were found between the number of lymphocytes in the blood with the markers CD5+, CD19+, CD23+, CD43+ and the T-score, Z-score, and BMD in the FNA. The data obtained are a prerequisite for further study of the interaction between the clone of tumor lymphocytes in CLL and cells involved in bone tissue remodeling to identify the mechanism of development of OP, osteoporosis and the risk of bone fractures, and changes in the quantitative composition of the lymphocyte phenotype in the blood can be considered as potential laboratory predictors of a decrease in BMD.
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Prognostic significance of hepatocyte growth factor in non-metastatic colorectal cancer
Abstract
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.
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Dental anomalies in congenital error of immunity
Abstract
Developmental disorders of the teeth and maxillofacial region are part of a symptoms complex that often occur with inborn errors of the immune system. The purpose of the study is to identify the frequency of various dentofacial anomalies occurrence in patients with diagnosed immunodeficiencies.
The study involved 64 patients of the Sverdlovsk Regional Clinical Hospital No. 1 and the Regional Children’s Clinical Hospital (Ekaterinburg) with various diagnoses: combined immunodeficiencies, antibody defects, autoinflammatory disorders, defects in the number and functions of phagocytes, defects in immune dysregulation, complement defects, and unverified diagnosis with signs of immunodeficiency. All patients underwent examination of the oral cavity, hard dental tissues, periodontal tissue and jaw bone tissue.
It was found that an anomaly in jaw size is characteristic of all studied patient groups, except for the group with complement defects. Along with this, such pathology as quantitative and qualitative changes in teeth was identified in four classification groups: “combined immunodeficiencies” (25% of cases from the subgroup), “antibody defects” (31% of cases from the subgroup), “phagocyte defects” (10 % of cases from the subgroup), and “immune dysregulation defects” (50% of cases from the subgroup). The same frequency of occurrence of tissue repair defects: in “combined immunodeficiencies” (16.6% of cases from the subgroup), “autoinflammatory disorders” (26.7% of cases from the subgroup), “phagocyte defects” (10% of cases from the subgroup), in “unverified diagnoses” (20% of cases from the subgroup). The highest percentage of quantitative manifestations of such anomalies as jaw size (26% of cases from the entire study group) and facial dysmorphism (27% of cases from the entire study group). In patients with “combined immunodeficiencies”, all the dental anomalies described in the work were identified. However, the highest percentage of people in this group had delayed teething (33.3%) and facial dysmorphism (33%). The only patient diagnosed with “complement defects” had no dental anomalies. The minimum number of manifestations of the studied anomalies was observed in groups with “antibody defects” and “immune dysregulation defects”.
The accumulated experience in describing various dental diagnoses will contribute to the creation of dental criteria for the diagnosis of primary immunodeficiencies.
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Immunology of the “sensible world” and of the “intelligible world”: the evolutionary decline and the “constructed reality”
Abstract
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 18th 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.
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