Pub Date : 2022-09-20DOI: 10.46235/1028-7221-1149-lmo
Y.A. Sarycheva, A. A. Tokareva, A. G. Shechtman, T. Panfilova, Yu. S. Pimenova, R. Mitrofanov, B. Frolov
Antimutagenic effect of the plant triterpenoid miliacin was studied, in order to characterize its protective properties in a model of acute irradiation immunosuppression using outbred male mice. Ionizing irradiation at different doses (0.5; 1.0; 2.0; 4.0 Gy) was used for experimental (miliacin-treated), and control animals that received the miliacin solvent. Miliacin was administered three times intraperitoneally at a single dose of 4.0 mg/kg with 24-hour intervals between injections. The last dose was applied 1 day before irradiation. Myelokaryocytes served as test objects, the analysis of which was carried out 24 hours after irradiation. Miliacin had a certain protective effect by limiting the post-radiation myeloablation, reducing the number of aberrant cells and the total number of aberrations. Protective effect of triterpenoids showed inverse relation to the radiation dose, being most pronounced at the dose of 0.5 Gy. Higher values of chromatid aberrations at radiation doses of 1.0 and 2.0 Gy in animals from the experimental group versus control mice, probably, due to anti-apoptotic effect of the triterpenoid, thus ensuring higher survival rates of mutated cells with severe damage to their genome. The protective effect of miliacin at 24 hours after radiation exposure may indicate its effect on the primary radiochemical stage of radiation injury. It is suggested that the mechanism of protective action of triterpenoid is mediated by its previously shown antioxidant activity, due to its ability to stabilize membranes and normalize expression of genes encoding antioxidant protection enzymes. Thus, the antimutagenic activity of miliacin after irradiation is an important characteristic of its immunoprotective effect during the radiation-induced immunosuppression. With respect to its ability to limit the mutagenic effect, miliacin may be classified as a weak radioprotective antimutagen with a protection efficiency of 20-40% at the dose range of 0.5 to 1.0 Gray.
{"title":"Limited mutagenesis of myelokaryocytes following acute external irradiation as a protective mechanism of miliacin in radiation-induced immunosuppression","authors":"Y.A. Sarycheva, A. A. Tokareva, A. G. Shechtman, T. Panfilova, Yu. S. Pimenova, R. Mitrofanov, B. Frolov","doi":"10.46235/1028-7221-1149-lmo","DOIUrl":"https://doi.org/10.46235/1028-7221-1149-lmo","url":null,"abstract":"Antimutagenic effect of the plant triterpenoid miliacin was studied, in order to characterize its protective properties in a model of acute irradiation immunosuppression using outbred male mice. Ionizing irradiation at different doses (0.5; 1.0; 2.0; 4.0 Gy) was used for experimental (miliacin-treated), and control animals that received the miliacin solvent. Miliacin was administered three times intraperitoneally at a single dose of 4.0 mg/kg with 24-hour intervals between injections. The last dose was applied 1 day before irradiation. Myelokaryocytes served as test objects, the analysis of which was carried out 24 hours after irradiation. Miliacin had a certain protective effect by limiting the post-radiation myeloablation, reducing the number of aberrant cells and the total number of aberrations. Protective effect of triterpenoids showed inverse relation to the radiation dose, being most pronounced at the dose of 0.5 Gy. Higher values of chromatid aberrations at radiation doses of 1.0 and 2.0 Gy in animals from the experimental group versus control mice, probably, due to anti-apoptotic effect of the triterpenoid, thus ensuring higher survival rates of mutated cells with severe damage to their genome. The protective effect of miliacin at 24 hours after radiation exposure may indicate its effect on the primary radiochemical stage of radiation injury. It is suggested that the mechanism of protective action of triterpenoid is mediated by its previously shown antioxidant activity, due to its ability to stabilize membranes and normalize expression of genes encoding antioxidant protection enzymes. Thus, the antimutagenic activity of miliacin after irradiation is an important characteristic of its immunoprotective effect during the radiation-induced immunosuppression. With respect to its ability to limit the mutagenic effect, miliacin may be classified as a weak radioprotective antimutagen with a protection efficiency of 20-40% at the dose range of 0.5 to 1.0 Gray.","PeriodicalId":21524,"journal":{"name":"Russian Journal of Immunology","volume":"7 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75539940","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-09-20DOI: 10.46235/1028-7221-1103-ira
E. Starostina, S. V. Sharabrin, A. Rudometov, V. R. Litvinova, M. B. Borgoyakova, S. Bazhan, A. Ilyichev, L. Karpenko
Constant antigenic drift of circulating influenza viruses leads to inefficiency of seasonal influenza vaccines, thus requiring annual re-design of these vaccines. Therefore, the development of a universal influenza vaccine is of particular relevance. A promising line of research in this area is to design the immunogens consisting of conserved protein fragments from different influenza viral strains. The aim of this work was to assess immunogenicity of DNA vaccines and mRNA vaccines encoding artificial antigens consisting of conserved hemagglutinin stem fragments and conserved M2 protein. We have obtained DNA vaccine constructs encoding artificial immunogens AgH1, AgH3, and AgM2, which contained conserved fragments of the hemagglutinin stalk from the two subtypes of influenza A H1N1 and H3N2, and conserved M2 protein. These DNA vaccines were used as templates for the synthesis of mRNA vaccines. To assess immunogenicity of the obtained constructs, BALB/c mice were immunized with DNA and mRNA vaccines by i/m administration. Assessment of the humoral immune response was carried out by ELISA, using influenza viruses A/Aichi/2/68(H3N2), A/California/07/2009 as antigens and the ULTRIX vaccine containing purified antigens of H1N1 and H3N2 influenza viruses. T cell immune response was assessed using two methods: intracellular cytokine staining (ICS) and ELISpot. ICS was performed to determine CD8+ and CD4+T-lymphocytes producing IFN. ELISpot was carried out using the mouse IFN ELISpot kit (BD). A peptide mixture which included composition of the target antigens, was used for cell stimulation. The results showed that the designed DNA vaccine constructs induce virus-specific humoral and cellular responses in immunized BALB/c mice. Intramuscular administration of the naked mRNA vaccine constructs induced a weak humoral immune response, thus suggesting a need for further work to improve the delivery approaches.
{"title":"Immune response against DNA- and mRNA vaccines encoding artificial influenza virus immunogens","authors":"E. Starostina, S. V. Sharabrin, A. Rudometov, V. R. Litvinova, M. B. Borgoyakova, S. Bazhan, A. Ilyichev, L. Karpenko","doi":"10.46235/1028-7221-1103-ira","DOIUrl":"https://doi.org/10.46235/1028-7221-1103-ira","url":null,"abstract":"Constant antigenic drift of circulating influenza viruses leads to inefficiency of seasonal influenza vaccines, thus requiring annual re-design of these vaccines. Therefore, the development of a universal influenza vaccine is of particular relevance. A promising line of research in this area is to design the immunogens consisting of conserved protein fragments from different influenza viral strains. The aim of this work was to assess immunogenicity of DNA vaccines and mRNA vaccines encoding artificial antigens consisting of conserved hemagglutinin stem fragments and conserved M2 protein. We have obtained DNA vaccine constructs encoding artificial immunogens AgH1, AgH3, and AgM2, which contained conserved fragments of the hemagglutinin stalk from the two subtypes of influenza A H1N1 and H3N2, and conserved M2 protein. These DNA vaccines were used as templates for the synthesis of mRNA vaccines. To assess immunogenicity of the obtained constructs, BALB/c mice were immunized with DNA and mRNA vaccines by i/m administration. Assessment of the humoral immune response was carried out by ELISA, using influenza viruses A/Aichi/2/68(H3N2), A/California/07/2009 as antigens and the ULTRIX vaccine containing purified antigens of H1N1 and H3N2 influenza viruses. T cell immune response was assessed using two methods: intracellular cytokine staining (ICS) and ELISpot. ICS was performed to determine CD8+ and CD4+T-lymphocytes producing IFN. ELISpot was carried out using the mouse IFN ELISpot kit (BD). A peptide mixture which included composition of the target antigens, was used for cell stimulation. The results showed that the designed DNA vaccine constructs induce virus-specific humoral and cellular responses in immunized BALB/c mice. Intramuscular administration of the naked mRNA vaccine constructs induced a weak humoral immune response, thus suggesting a need for further work to improve the delivery approaches.","PeriodicalId":21524,"journal":{"name":"Russian Journal of Immunology","volume":"34 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88409628","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-09-20DOI: 10.46235/1028-7221-1152-ceo
M. Osikov, N. V. Kaigorodtseva, M. Boyko, A. Fedosov
Multiple effects of ozone are a prerequisite for its use in the complex therapy of inflammatory bowel diseases. Our aim was to analyze the effects of ozone and 5-aminosalicylic acid (5-ASA) in oxazolone-induced colitis (OIC) upon the innate immunity functions. OIC was modeled in 72 male Wistar rats weighing 24020 g by a two-stage application of oxazolone solution. Ozone at a dose of 0.05 mg/kg, as a part of ozone-oxygen mixture, was injected intraperitoneally every 24 hours. The laboratory indices were studied on the days 2, 4, 6. In blood samples, the number of leukocytes, differential leukocyte counts, absorptive capacity of blood neutrophils and their oxygen-dependent metabolism were studied. In homogenate of the colon mucosa, IL-17 concentration was measured. In OIC, on days 2, 4, and 6, the number of blood leukocytes was increased, with a predominance of lymphocytes, monocytes, neutrophils and their absorption capacity; the concentration of IL-17 increased in the areas of colonic damage. On days 2 and 4, the activity and intensity of the spontaneous NBT test is increased like as activity and intensity of induced NBT-test on days 2 and 6. Administration of ozone reduced the content of blood leukocytes, lymphocytes, neutrophils on days 2 and 6, like as their absorption capacity on days 2 and 4, along with a decrease in NCT-reducing ability on days 6, and decreased IL-17 concentration in the area of colonic damage on days 4 and 6. The effects of ozone administration in OIC, if compared with 5-ASA, are less pronounced on days 2 and 4, with respect to decreased number of monocytes, neutrophils in blood and their absorption capacity on days 2, like as IL-17 concentration in the area of colonic damage on days 4 and 6.
{"title":"Comparative effect of ozone exposure and 5-aminosalicylic acid in oxazolon-induced colitis upon the indices of innate immunity","authors":"M. Osikov, N. V. Kaigorodtseva, M. Boyko, A. Fedosov","doi":"10.46235/1028-7221-1152-ceo","DOIUrl":"https://doi.org/10.46235/1028-7221-1152-ceo","url":null,"abstract":"Multiple effects of ozone are a prerequisite for its use in the complex therapy of inflammatory bowel diseases. Our aim was to analyze the effects of ozone and 5-aminosalicylic acid (5-ASA) in oxazolone-induced colitis (OIC) upon the innate immunity functions. \u0000OIC was modeled in 72 male Wistar rats weighing 24020 g by a two-stage application of oxazolone solution. Ozone at a dose of 0.05 mg/kg, as a part of ozone-oxygen mixture, was injected intraperitoneally every 24 hours. The laboratory indices were studied on the days 2, 4, 6. In blood samples, the number of leukocytes, differential leukocyte counts, absorptive capacity of blood neutrophils and their oxygen-dependent metabolism were studied. In homogenate of the colon mucosa, IL-17 concentration was measured. \u0000In OIC, on days 2, 4, and 6, the number of blood leukocytes was increased, with a predominance of lymphocytes, monocytes, neutrophils and their absorption capacity; the concentration of IL-17 increased in the areas of colonic damage. On days 2 and 4, the activity and intensity of the spontaneous NBT test is increased like as activity and intensity of induced NBT-test on days 2 and 6. Administration of ozone reduced the content of blood leukocytes, lymphocytes, neutrophils on days 2 and 6, like as their absorption capacity on days 2 and 4, along with a decrease in NCT-reducing ability on days 6, and decreased IL-17 concentration in the area of colonic damage on days 4 and 6. The effects of ozone administration in OIC, if compared with 5-ASA, are less pronounced on days 2 and 4, with respect to decreased number of monocytes, neutrophils in blood and their absorption capacity on days 2, like as IL-17 concentration in the area of colonic damage on days 4 and 6.","PeriodicalId":21524,"journal":{"name":"Russian Journal of Immunology","volume":"34 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87082891","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-09-20DOI: 10.46235/1028-7221-1117-foi
I. D. Magamedov, L. Pivovarova, S. P. Nokhrin, V. Soroka, O. Ariskina, I. V. Osipova, I. M. Radjabov, K. Fomin, S. L. Potskhor-ogly, L. V. Kolichenko, E. Markelova, O. Goncharova
Lymphocyte-to-platelet adhesion during hypoxia, tissue damage, activation of inflammation and coagulation is associated with expression of ICAM-1 membrane molecules by blood and tissue cells. At the same time, the platelet adhesion receptors determine their adherence to endothelium and recruited lymphocytes. Moreover, the role of platelets in pathogenesis of ischemic cardiovascular diseases comprises their ability to modulate both hemostasis and inflammatory reactions, which are accompanied by secretion of inflammatory mediators and some factors that promote recruitment of leukocytes to tissue damage sites. Creatine kinase activity is a sensitive marker of tissue damage and tissue ischemia. The purpose of the present study was to assess the effect of anti-inflammatory therapy with dexamethasone upon the intensity of inflammation and adhesive properties of lymphocytes, number of platelets in peripheral blood of the patients with acute lower limb ischemia (ALLI), as well as to evaluate the effectiveness of treatment. To study the effect of anti-inflammatory therapy, a group of 32 patients treated with dexamethasone was selected; the control group was represented by 71 patients with basic therapy, the comparison group consisted of 15 volunteers. After revascularization, all patients received antiplatelet and anticoagulant therapy. Dexamethasone infusions were carried out as a course of 4 to 6 days after reconstructive surgery. In all patients, the content of C-reactive protein in blood, the activity of creatine kinase, the content of platelets and, especially, of enlarged platelets were determined. The numbers of lymphocytes expressing ICAM-1 (CD54+) adhesion molecules were counted using immunocytochemical technique. The studies were performed before surgery and 1, 3, 5, 7, 10 days after surgery. During exacerbation of the limb ischemia and damage to endothelium, the accumulation of cytolysis products was noted. Expression of adhesion molecules was increased both on endotheliocytes and on inflammation effector cells, i.e., leukocytes and platelets. The adhesion molecules transmit the activating signal inside the cell, thus promoting adhesion of leukocytes and platelets to endothelium, lymphocytic-platelet adhesion, formation of parietal thrombi, and possible occlusion of damaged vessels. Increased expression of adhesion molecules is associated with activation of metabolism, inflammation, coagulation and oxidative stress. It may stimulate all hematopoietic lineages, including platelets. The involvement level of cellular reactions in pathogenesis of the disease affects effectiveness and duration of treatment, risk of recurrent thrombosis and lethal outcome. Anti-inflammatory therapy with dexamethasone contributed to earlier remission, it was associated with lower frequency of infectious and thrombotic complications, decreased mortality, and reduced duration of treatment. Inflammation, adhesiveness of effector cells and thrombosis are important fact
{"title":"Factors of inflammatory, adhesiveness and thrombosis in acute lower limb ischemia and dexamethasone therapy","authors":"I. D. Magamedov, L. Pivovarova, S. P. Nokhrin, V. Soroka, O. Ariskina, I. V. Osipova, I. M. Radjabov, K. Fomin, S. L. Potskhor-ogly, L. V. Kolichenko, E. Markelova, O. Goncharova","doi":"10.46235/1028-7221-1117-foi","DOIUrl":"https://doi.org/10.46235/1028-7221-1117-foi","url":null,"abstract":"Lymphocyte-to-platelet adhesion during hypoxia, tissue damage, activation of inflammation and coagulation is associated with expression of ICAM-1 membrane molecules by blood and tissue cells. At the same time, the platelet adhesion receptors determine their adherence to endothelium and recruited lymphocytes. Moreover, the role of platelets in pathogenesis of ischemic cardiovascular diseases comprises their ability to modulate both hemostasis and inflammatory reactions, which are accompanied by secretion of inflammatory mediators and some factors that promote recruitment of leukocytes to tissue damage sites. Creatine kinase activity is a sensitive marker of tissue damage and tissue ischemia. The purpose of the present study was to assess the effect of anti-inflammatory therapy with dexamethasone upon the intensity of inflammation and adhesive properties of lymphocytes, number of platelets in peripheral blood of the patients with acute lower limb ischemia (ALLI), as well as to evaluate the effectiveness of treatment. \u0000To study the effect of anti-inflammatory therapy, a group of 32 patients treated with dexamethasone was selected; the control group was represented by 71 patients with basic therapy, the comparison group consisted of 15 volunteers. After revascularization, all patients received antiplatelet and anticoagulant therapy. Dexamethasone infusions were carried out as a course of 4 to 6 days after reconstructive surgery. In all patients, the content of C-reactive protein in blood, the activity of creatine kinase, the content of platelets and, especially, of enlarged platelets were determined. The numbers of lymphocytes expressing ICAM-1 (CD54+) adhesion molecules were counted using immunocytochemical technique. The studies were performed before surgery and 1, 3, 5, 7, 10 days after surgery. \u0000During exacerbation of the limb ischemia and damage to endothelium, the accumulation of cytolysis products was noted. Expression of adhesion molecules was increased both on endotheliocytes and on inflammation effector cells, i.e., leukocytes and platelets. The adhesion molecules transmit the activating signal inside the cell, thus promoting adhesion of leukocytes and platelets to endothelium, lymphocytic-platelet adhesion, formation of parietal thrombi, and possible occlusion of damaged vessels. Increased expression of adhesion molecules is associated with activation of metabolism, inflammation, coagulation and oxidative stress. It may stimulate all hematopoietic lineages, including platelets. The involvement level of cellular reactions in pathogenesis of the disease affects effectiveness and duration of treatment, risk of recurrent thrombosis and lethal outcome. Anti-inflammatory therapy with dexamethasone contributed to earlier remission, it was associated with lower frequency of infectious and thrombotic complications, decreased mortality, and reduced duration of treatment. \u0000Inflammation, adhesiveness of effector cells and thrombosis are important fact","PeriodicalId":21524,"journal":{"name":"Russian Journal of Immunology","volume":"38 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80979770","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-09-20DOI: 10.46235/1028-7221-1155-eoc
T. Radygina, D. Kuptsova, S. Petrichuk, E. Semikina, A. Fisenko
Purinergic system plays an important role in functional regulation of immune system. Extracellular ATP belongs to non-infectious danger signals (DAMP), has a pro-inflammatory effect and modulates the immune response. The end product of ATP breakdown, adenosine, plays an important role in limiting the inflammatory response. The activity of ectonucleotidase CD39 and CD73 supports the balance of ATP and adenosine at the site of inflammation. CD39 and CD73 expression is characterized by high variability. From the literature data, appropriate studies were carried out either in transgenic mice, or with adult healthy donors. The number of cells expressing CD39 and CD73 in T lymphocyte populations has not been evaluated in healthy children. Hence, our aim was to study the features of CD39 and CD73 expression in various subpopulations of CD4+ lymphocytes in apparently healthy children of different ages. We examined 45 healthy children aged 3.7 to 17.5 years (Me (Q0.25-Q0.75) 12.4 (10-16.1). The numbers of CD4+ cells, regulatory T lymphocytes (Treg CD4+CD25highCD127low), activated T helpers (Tact CD4+CD25+CD127high), and Th17 lymphocytes (CD4+CD161+CD3+) expressing CD39 and CD73 were evaluated by the flow cytometry. The number of cells expressing CD39 and CD73 depends on specific cell subpopulation. The highest content of CD39+ cells was observed in Tregs, and maximal amounts of CD73+ cells were found among Tact subset. In the Th17 lymphocyte subpopulation, there was no significant difference between the number of cells expressing CD39 and CD73. E have also shown an increase in the relative number of Th17 cells expressing CD73, along with age-dependent decrease in the relative number of Tact cells expressing CD39. An age-dependent decrease in absolute values with age was revealed for Treg, CD39+Treg, CD73+Treg, CD39+Th17, thus being consistent with age-related decrease in absolute numbers of lymphocytes and CD4+ cells. The obtained data concerning specific pattern of ectonucleotidases expression in functionally different populations of CD4+ lymphocytes should be taken into account when studying children with immune-mediated diseases from different age groups.
{"title":"Expression of CD39 and CD73 ectonucleotidases in CD4+ lymphocyte populations in healthy children","authors":"T. Radygina, D. Kuptsova, S. Petrichuk, E. Semikina, A. Fisenko","doi":"10.46235/1028-7221-1155-eoc","DOIUrl":"https://doi.org/10.46235/1028-7221-1155-eoc","url":null,"abstract":"Purinergic system plays an important role in functional regulation of immune system. Extracellular ATP belongs to non-infectious danger signals (DAMP), has a pro-inflammatory effect and modulates the immune response. The end product of ATP breakdown, adenosine, plays an important role in limiting the inflammatory response. The activity of ectonucleotidase CD39 and CD73 supports the balance of ATP and adenosine at the site of inflammation. CD39 and CD73 expression is characterized by high variability. From the literature data, appropriate studies were carried out either in transgenic mice, or with adult healthy donors. The number of cells expressing CD39 and CD73 in T lymphocyte populations has not been evaluated in healthy children. Hence, our aim was to study the features of CD39 and CD73 expression in various subpopulations of CD4+ lymphocytes in apparently healthy children of different ages. \u0000We examined 45 healthy children aged 3.7 to 17.5 years (Me (Q0.25-Q0.75) 12.4 (10-16.1). The numbers of CD4+ cells, regulatory T lymphocytes (Treg CD4+CD25highCD127low), activated T helpers (Tact CD4+CD25+CD127high), and Th17 lymphocytes (CD4+CD161+CD3+) expressing CD39 and CD73 were evaluated by the flow cytometry. \u0000The number of cells expressing CD39 and CD73 depends on specific cell subpopulation. The highest content of CD39+ cells was observed in Tregs, and maximal amounts of CD73+ cells were found among Tact subset. In the Th17 lymphocyte subpopulation, there was no significant difference between the number of cells expressing CD39 and CD73. E have also shown an increase in the relative number of Th17 cells expressing CD73, along with age-dependent decrease in the relative number of Tact cells expressing CD39. An age-dependent decrease in absolute values with age was revealed for Treg, CD39+Treg, CD73+Treg, CD39+Th17, thus being consistent with age-related decrease in absolute numbers of lymphocytes and CD4+ cells. \u0000The obtained data concerning specific pattern of ectonucleotidases expression in functionally different populations of CD4+ lymphocytes should be taken into account when studying children with immune-mediated diseases from different age groups.","PeriodicalId":21524,"journal":{"name":"Russian Journal of Immunology","volume":"38 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74033335","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-09-20DOI: 10.46235/1028-7221-1139-cao
D. Stashkevich, S. Belyaeva, A. V. Evdokimov
Ulcerative colitis and irritable bowel syndrome are multifactorial disorders with genetic predisposition. Recent studies suggest that the mucosal immune activation, increased intestinal permeability, and altered host-microbiota interactions may modulate innate immune response, thus contributing to immunopathogenesis of these diseases. Toll-like receptors (TLR) are considered to play the main role in genetic susceptibility to the conditions. The mechanisms for regulating activity of Toll-like receptors are represented by single-nucleotide gene polymorphisms (SNPs), thus producing allelotypes with different biological effects. Among all known TLRs, TLR2 is the most actively studied. The TLR2 gene is located on the long arm of the chromosome 4 and contains the genetic variant leading to the substitution of arginine for glutamine (Arg753Gln) in TLR2 protein. Meanwhile, the most studied SNP of TLR6 is located at the C745T position causing Pro249Ser amino acid substitution in the protein. The present work aimed for analysis of distribution of alleles, genotypes and haplotype combinations of the TLR2 and TLR6 SNPs, and their associations with predisposal for ulcerative colitis and irritable bowel syndrome in Russians from Chelyabinsk Region. The following methods were used in the study: isolation of DNA samples from whole blood, genotyping of the studied gene polymorphisms using PCR with electrophoretic detection. The frequencies of two-locus haplotypes formed by SNPs TLR2 TLR6 were calculated with Arlequin ver 3.5 software. Comparison of two populational samples for predisposition to UC and IBS was carried out using standard immunogenetic criteria. Significance of differences was set at p 0.05. Results: Analysis of the data showed the association of specific alleles and genotypes, but not TLR2 TLR6 haplotypes, with predisposition to the studied diseases. The Arg753Gln gene polymorphism of TLR2 was shown to be significant for a predisposition to ulcerative colitis, and SNP Pro249Ser TLR6 is associated with susceptibility to irritable bowel syndrome in Russians from the Chelyabinsk Region.
{"title":"Comparative assessment of genetic polymorphism of Toll-like 2 and 6 receptors predisposing for non-specific ulcerative colitis and irritable bowel syndrome in Russians from Chelyabinsk Region","authors":"D. Stashkevich, S. Belyaeva, A. V. Evdokimov","doi":"10.46235/1028-7221-1139-cao","DOIUrl":"https://doi.org/10.46235/1028-7221-1139-cao","url":null,"abstract":"Ulcerative colitis and irritable bowel syndrome are multifactorial disorders with genetic predisposition. Recent studies suggest that the mucosal immune activation, increased intestinal permeability, and altered host-microbiota interactions may modulate innate immune response, thus contributing to immunopathogenesis of these diseases. Toll-like receptors (TLR) are considered to play the main role in genetic susceptibility to the conditions. The mechanisms for regulating activity of Toll-like receptors are represented by single-nucleotide gene polymorphisms (SNPs), thus producing allelotypes with different biological effects. Among all known TLRs, TLR2 is the most actively studied. The TLR2 gene is located on the long arm of the chromosome 4 and contains the genetic variant leading to the substitution of arginine for glutamine (Arg753Gln) in TLR2 protein. Meanwhile, the most studied SNP of TLR6 is located at the C745T position causing Pro249Ser amino acid substitution in the protein. The present work aimed for analysis of distribution of alleles, genotypes and haplotype combinations of the TLR2 and TLR6 SNPs, and their associations with predisposal for ulcerative colitis and irritable bowel syndrome in Russians from Chelyabinsk Region. The following methods were used in the study: isolation of DNA samples from whole blood, genotyping of the studied gene polymorphisms using PCR with electrophoretic detection. The frequencies of two-locus haplotypes formed by SNPs TLR2 TLR6 were calculated with Arlequin ver 3.5 software. Comparison of two populational samples for predisposition to UC and IBS was carried out using standard immunogenetic criteria. Significance of differences was set at p 0.05. Results: Analysis of the data showed the association of specific alleles and genotypes, but not TLR2 TLR6 haplotypes, with predisposition to the studied diseases. The Arg753Gln gene polymorphism of TLR2 was shown to be significant for a predisposition to ulcerative colitis, and SNP Pro249Ser TLR6 is associated with susceptibility to irritable bowel syndrome in Russians from the Chelyabinsk Region.","PeriodicalId":21524,"journal":{"name":"Russian Journal of Immunology","volume":"58 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"77308094","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-09-20DOI: 10.46235/1028-7221-1148-eog
S. S. Lazarev, M. Bochkova, V. Timganova, M. Rayev
Graphene oxide (GO) is a promising material, which is likely to find applications in the fields of medicine and biotechnology. However, the current knowledge of its influence on human organism is limited. Even less information is available on the effects of GO on the cell lines widely used in biotechnology. The aim of this work is to describe the interaction between GO nanoparticles and BAP3 hybridoma cells which produce anti-human-PSG1 IgG, in vitro. We studied the effect of GO nanoparticles on cell viability and the intensity of internalization (adhesion) of nanoparticles by the cells. We used GO nanoparticles of different size, with surface being functionalized by linear or branched PEG (GO-PEG). The PEG coating level was 20% (by mass). The following nanoparticle concentrations were used: 5 g/mL and 25 g/mL. The BAP3 cells were cultured in a 48-well cell culture plates in serum-free DCCM-1 media in the presence of GO nanoparticles. The cells were cultured for 24 hours at 37 С and 5% СО2. Cell viability was assessed by a flow cytometer utilizing Zombie Aqua (ZA) staining. Internalization (adhesion) of nanoparticles was monitored using a flow cytometer by GO fluorescense in the samples (ex = 488 nm). Moreover, interactions between hybridoma cells and GO nanoparticles were visualized by EVOS M5000 visualization system, which included an inverted fluorescent microscope. We demonstrated that GO nanoparticles possess a cytotoxic effect when applied at high concentration (25 g/mL). The highest cytotoxic effect is caused by GO nanoparticles coated with linear PEG. The degree of nanoparticle internalization (adhesion) was shown to be significantly lower when the particles were present at lower (5 g/mL) concentration. Internalization (adhesion) of nanoparticles of smaller size was more abundant. Furthermore, these nanoparticles were shown to have a stronger cytotoxic effect compared to larger particles. In general, cytotoxicity of GO nanoparticles decreases with increasing size, which is especially evident if the fact that the mean effective diameter of the nanoparticles coated with branched PEG is considered larger than their linear PEG-coated counterparts. The data obtained allow us to draw a correlation between the cytotoxic effect of GO nanoparticles and the level of their internalization (adhesion) by the cells. In general, this work concerns some novel aspects of interaction between GO nanoparticles and hybridoma cells.
{"title":"Effect of graphene oxide nanoparticles on viability of BAP3 hybridoma cells","authors":"S. S. Lazarev, M. Bochkova, V. Timganova, M. Rayev","doi":"10.46235/1028-7221-1148-eog","DOIUrl":"https://doi.org/10.46235/1028-7221-1148-eog","url":null,"abstract":"Graphene oxide (GO) is a promising material, which is likely to find applications in the fields of medicine and biotechnology. However, the current knowledge of its influence on human organism is limited. Even less information is available on the effects of GO on the cell lines widely used in biotechnology. The aim of this work is to describe the interaction between GO nanoparticles and BAP3 hybridoma cells which produce anti-human-PSG1 IgG, in vitro. We studied the effect of GO nanoparticles on cell viability and the intensity of internalization (adhesion) of nanoparticles by the cells. We used GO nanoparticles of different size, with surface being functionalized by linear or branched PEG (GO-PEG). The PEG coating level was 20% (by mass). The following nanoparticle concentrations were used: 5 g/mL and 25 g/mL. The BAP3 cells were cultured in a 48-well cell culture plates in serum-free DCCM-1 media in the presence of GO nanoparticles. The cells were cultured for 24 hours at 37 С and 5% СО2. Cell viability was assessed by a flow cytometer utilizing Zombie Aqua (ZA) staining. Internalization (adhesion) of nanoparticles was monitored using a flow cytometer by GO fluorescense in the samples (ex = 488 nm). Moreover, interactions between hybridoma cells and GO nanoparticles were visualized by EVOS M5000 visualization system, which included an inverted fluorescent microscope. \u0000We demonstrated that GO nanoparticles possess a cytotoxic effect when applied at high concentration (25 g/mL). The highest cytotoxic effect is caused by GO nanoparticles coated with linear PEG. The degree of nanoparticle internalization (adhesion) was shown to be significantly lower when the particles were present at lower (5 g/mL) concentration. Internalization (adhesion) of nanoparticles of smaller size was more abundant. Furthermore, these nanoparticles were shown to have a stronger cytotoxic effect compared to larger particles. In general, cytotoxicity of GO nanoparticles decreases with increasing size, which is especially evident if the fact that the mean effective diameter of the nanoparticles coated with branched PEG is considered larger than their linear PEG-coated counterparts. The data obtained allow us to draw a correlation between the cytotoxic effect of GO nanoparticles and the level of their internalization (adhesion) by the cells. In general, this work concerns some novel aspects of interaction between GO nanoparticles and hybridoma cells.","PeriodicalId":21524,"journal":{"name":"Russian Journal of Immunology","volume":"57 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84842975","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-09-01DOI: 10.46235/1028-7221-1112-rbc
T. A. Bondarenko, E. Ivanova, A. V. Bekpergenova, I. N. Chaynikova, O. E. Chelpachenko, Igor A. Nikiforov, I. A. Zdvizhkova
Cytokines and chemokines, as well as gut microsymbionts, are sufficient participants in the intercellular communications, thus supporting homeostasis of gut mucosa. However, these components may be of key significance for intestinal inflammation and damage to epithelial barrier. This work expands the understanding of the relationships between intestinal microbial communities and the local cytokine network of the host. The paper presents the results of the correlation analysis between total microbial number of intestinal microsymbionts and the level of pro- (TNF, IFN, IL-8) and anti-inflammatory cytokines (IL-10, IL- 1ra) in coprofiltrates obtained from clinically healthy people examined for gut dysbiosis. Determination of cytokines in coprofiltrates was carried out by ELISA technique (JSC Vector-Best, Russia). The study of 65 microsymbiocenoses of the human gut was carried out by classical bacteriological methods. Identification of obligate-anaerobic, facultative-anaerobic bacteria and fungi was carried out by time-of-flight mass spectrometry using MALDI TOF-MS Microflex LT series (Bruker Daltonians, Germany). These studies have revealed the leading role of associations between enterobacteria, fungi and representatives of the Staphylococcus genus in gut dysbiosis. In general composition of the obligate-anaerobic association, we have observed a change of consortia from several types of Bifidobacteria and Lactobacilli in eubiotic state to a monoid variant in dysbiosis. At the same time, the number of associations that included Clostridia was increased. The analysis of correlations between cytokine indices and the number of gut microbiota showed persistance of significant associations established during eubiosis under dysbiosis conditions, with an increase in their correlation coefficient: Bifidobacterium spp., Enterobacteriaceae, Staphylococcus spp., Candida spp. and TNF. At the same time, in dysbiosis, the direction of the connections changed, and new correlations were determined: for Staphylococcus spp. and IFN; Staphylococcus spp. and IL-8; Enterobacteriaceae and IL-1ra, IFN. The established features of correlations between indices of microsymbiocenosis and quantitative changes in cytokines allow us to consider the number, composition of microsymbiocenosis and cytokine profile as factors that may affect the state of gut homeostasis in eu- and dysbiosis.
{"title":"Relationships between cytokines and the amounts of microsymbionts in microecological disorders of the human intestine","authors":"T. A. Bondarenko, E. Ivanova, A. V. Bekpergenova, I. N. Chaynikova, O. E. Chelpachenko, Igor A. Nikiforov, I. A. Zdvizhkova","doi":"10.46235/1028-7221-1112-rbc","DOIUrl":"https://doi.org/10.46235/1028-7221-1112-rbc","url":null,"abstract":"Cytokines and chemokines, as well as gut microsymbionts, are sufficient participants in the intercellular communications, thus supporting homeostasis of gut mucosa. However, these components may be of key significance for intestinal inflammation and damage to epithelial barrier. This work expands the understanding of the relationships between intestinal microbial communities and the local cytokine network of the host. The paper presents the results of the correlation analysis between total microbial number of intestinal microsymbionts and the level of pro- (TNF, IFN, IL-8) and anti-inflammatory cytokines (IL-10, IL- 1ra) in coprofiltrates obtained from clinically healthy people examined for gut dysbiosis. Determination of cytokines in coprofiltrates was carried out by ELISA technique (JSC Vector-Best, Russia). The study of 65 microsymbiocenoses of the human gut was carried out by classical bacteriological methods. Identification of obligate-anaerobic, facultative-anaerobic bacteria and fungi was carried out by time-of-flight mass spectrometry using MALDI TOF-MS Microflex LT series (Bruker Daltonians, Germany). These studies have revealed the leading role of associations between enterobacteria, fungi and representatives of the Staphylococcus genus in gut dysbiosis. In general composition of the obligate-anaerobic association, we have observed a change of consortia from several types of Bifidobacteria and Lactobacilli in eubiotic state to a monoid variant in dysbiosis. At the same time, the number of associations that included Clostridia was increased. The analysis of correlations between cytokine indices and the number of gut microbiota showed persistance of significant associations established during eubiosis under dysbiosis conditions, with an increase in their correlation coefficient: Bifidobacterium spp., Enterobacteriaceae, Staphylococcus spp., Candida spp. and TNF. At the same time, in dysbiosis, the direction of the connections changed, and new correlations were determined: for Staphylococcus spp. and IFN; Staphylococcus spp. and IL-8; Enterobacteriaceae and IL-1ra, IFN. The established features of correlations between indices of microsymbiocenosis and quantitative changes in cytokines allow us to consider the number, composition of microsymbiocenosis and cytokine profile as factors that may affect the state of gut homeostasis in eu- and dysbiosis.","PeriodicalId":21524,"journal":{"name":"Russian Journal of Immunology","volume":"29 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74064420","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-09-01DOI: 10.46235/1028-7221-1153-doc
I. Kritsky, V. Zurochka, Desheng Hu, A. Sarapultsev
Serological assays, being rapid and relatively inexpensive methods for detecting COVID-19, may play an important role in combating the SARS-CoV-2 pandemic. The aim of the present study was to assess dynamics of changes in the number of seropositive patients for SARS-CoV-2 antibodies over 2.5 years of the evolving COVID-19 pandemic. The study included 6051 persons (2840 women and 3211 males). Their mean age was 41.680.17 years (MSEM). At the time of this survey, all participants were residents of the Chelyabinsk region. General information was collected over the period from 06/01/2020 to 01/18/2022. Seropositivity for SARS-C0V-2 was assessed by test kits for IgG, IgM and IgA antibodies (JSC Vector-Best, Novosibirsk, Russia) against SARS-CoV-2 using indirect two-stage enzyme immunoassay (ELISA). Over the entire period, 27 cases were seronegative (20.45%); 99 samples were positive for IgA to SARS-CoV-2 (75%), and 6 samples (4.55%) yielded questionable ELISA results. IgG testing for SARS-Cov-2 antibodies was negative in 2433 cases (42.35%); 3245 samples (56.48%) were positive, and 67 specimens provided (1.17%) doubtful results using ELISA tests. IgM antibodies were not revealed in 2710 (70.41%) cases; 996 were positive (25.88%), and 143 specimens (3.72%) yielded doubtful results by ELISA technique. In general, the highest proportion of positive results was found in class A immunoglobulins. The wave-like distribution of the density among all antibody-positive patients was revealed, which, however, was not associated with peak values of COVID-19 morbidity in Chelyabinsk Region. Most waves of seroprevalence were detected before the waves of SARS-CoV-2 infection. A positive relationship was established between IgG and IgM seropositivity against SARS-CoV-2 with age and female gender. Conclusion. In general, serological testing and regular monitoring of antibodies against SARS-CoV-2 may play an important role in assessing its prevalence during the coronavirus pandemic and immune response to the infection at a population level.
{"title":"Dynamics of changes in the number of SARS-CoV-2 seropositive patients over two years of the COVID-19 pandemic","authors":"I. Kritsky, V. Zurochka, Desheng Hu, A. Sarapultsev","doi":"10.46235/1028-7221-1153-doc","DOIUrl":"https://doi.org/10.46235/1028-7221-1153-doc","url":null,"abstract":"Serological assays, being rapid and relatively inexpensive methods for detecting COVID-19, may play an important role in combating the SARS-CoV-2 pandemic. The aim of the present study was to assess dynamics of changes in the number of seropositive patients for SARS-CoV-2 antibodies over 2.5 years of the evolving COVID-19 pandemic. The study included 6051 persons (2840 women and 3211 males). Their mean age was 41.680.17 years (MSEM). At the time of this survey, all participants were residents of the Chelyabinsk region. General information was collected over the period from 06/01/2020 to 01/18/2022. Seropositivity for SARS-C0V-2 was assessed by test kits for IgG, IgM and IgA antibodies (JSC Vector-Best, Novosibirsk, Russia) against SARS-CoV-2 using indirect two-stage enzyme immunoassay (ELISA). Over the entire period, 27 cases were seronegative (20.45%); 99 samples were positive for IgA to SARS-CoV-2 (75%), and 6 samples (4.55%) yielded questionable ELISA results. IgG testing for SARS-Cov-2 antibodies was negative in 2433 cases (42.35%); 3245 samples (56.48%) were positive, and 67 specimens provided (1.17%) doubtful results using ELISA tests. IgM antibodies were not revealed in 2710 (70.41%) cases; 996 were positive (25.88%), and 143 specimens (3.72%) yielded doubtful results by ELISA technique. In general, the highest proportion of positive results was found in class A immunoglobulins. The wave-like distribution of the density among all antibody-positive patients was revealed, which, however, was not associated with peak values of COVID-19 morbidity in Chelyabinsk Region. Most waves of seroprevalence were detected before the waves of SARS-CoV-2 infection. A positive relationship was established between IgG and IgM seropositivity against SARS-CoV-2 with age and female gender. Conclusion. In general, serological testing and regular monitoring of antibodies against SARS-CoV-2 may play an important role in assessing its prevalence during the coronavirus pandemic and immune response to the infection at a population level.","PeriodicalId":21524,"journal":{"name":"Russian Journal of Immunology","volume":"187 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79759958","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-09-01DOI: 10.46235/1028-7221-1110-ped
E. D. Gavrilova, E. N. Demchenko, E. Goiman, O. Chumasova, N. Volskiy, A. Sizikov, V. Kozlov
Neutrophilic leukocytes play a key role for the joint damage in development of rheumatoid arthritis (RA). The specific death mode of these cells (netosis) may be an important reason of increase of cell-free DNA (cfDNA) in peripheral blood of the RA patients. Of great interest would be studies of alleged relationships between the of blood cfDNA contents being able of playing the role of an auto-antigen participating in the initiation of autoimmune reactions, and indices of neutrophil activation in this immunopathological disorder. The aim of the present study was to determine the levels of cfDNA in blood plasma of patients with RA depending on the clinical course of the disease, and to evaluate possible relationships between this index and activation of neutrophilic leukocytes. The study was conducted on 28 conditionally healthy donors and 63 patients with RA from the Rheumatology Department at the Clinic of Immunopathology (Novosibirsk). The level of cfDNA was determined using PicoGreen fluorescent dye. Neutrophils from the peripheral blood of donors and patients with rheumatoid arthritis were isolated in a Ficoll-Urografin density gradient. Neutrophilic leukocytes accounted for more than 98% of the fraction of isolated cells, and their viability was 99%. A portion of freshly isolated neutrophils was stimulated by phorbol myristate acetate. Concentration of myeloperoxidase in blood plasma of donors and patients with RA was determined using the Human MPO ELISA kit. It has been shown that the increased concentration of extracellular DNA in blood plasma of RA patients correlates with an higher degree of disease activity, and this parameter may serve as a relatively independent indicator of the disease intensity. A correlation was found between the level of cfDNA and common biochemical markers used to assess the activity of disease, i.e., DAS-28 and C-reactive protein levels in serum (p 0.05). Decrease of cfDNA concentrations is detected during treatment of the RA patients. This is due to the expected prognosis, i.e., a decreased manifestation of the disease, which also means correct administration of therapy. A relationship was found between the level of cfDNA and blood myeloperoxidase concentration in RA patients. The data obtained during the study suggest a possible connection between increased concentration of extracellular DNA, and activation of neutrophilic leukocytes in rheumatoid arthritis, with increased netosis in the affected joints.
{"title":"Plasma extracellular DNA and neutrophilic leukocyte activity in patients with rheumatoid arthritis","authors":"E. D. Gavrilova, E. N. Demchenko, E. Goiman, O. Chumasova, N. Volskiy, A. Sizikov, V. Kozlov","doi":"10.46235/1028-7221-1110-ped","DOIUrl":"https://doi.org/10.46235/1028-7221-1110-ped","url":null,"abstract":"Neutrophilic leukocytes play a key role for the joint damage in development of rheumatoid arthritis (RA). The specific death mode of these cells (netosis) may be an important reason of increase of cell-free DNA (cfDNA) in peripheral blood of the RA patients. Of great interest would be studies of alleged relationships between the of blood cfDNA contents being able of playing the role of an auto-antigen participating in the initiation of autoimmune reactions, and indices of neutrophil activation in this immunopathological disorder. The aim of the present study was to determine the levels of cfDNA in blood plasma of patients with RA depending on the clinical course of the disease, and to evaluate possible relationships between this index and activation of neutrophilic leukocytes. The study was conducted on 28 conditionally healthy donors and 63 patients with RA from the Rheumatology Department at the Clinic of Immunopathology (Novosibirsk). The level of cfDNA was determined using PicoGreen fluorescent dye. Neutrophils from the peripheral blood of donors and patients with rheumatoid arthritis were isolated in a Ficoll-Urografin density gradient. Neutrophilic leukocytes accounted for more than 98% of the fraction of isolated cells, and their viability was 99%. A portion of freshly isolated neutrophils was stimulated by phorbol myristate acetate. Concentration of myeloperoxidase in blood plasma of donors and patients with RA was determined using the Human MPO ELISA kit. It has been shown that the increased concentration of extracellular DNA in blood plasma of RA patients correlates with an higher degree of disease activity, and this parameter may serve as a relatively independent indicator of the disease intensity. A correlation was found between the level of cfDNA and common biochemical markers used to assess the activity of disease, i.e., DAS-28 and C-reactive protein levels in serum (p 0.05). Decrease of cfDNA concentrations is detected during treatment of the RA patients. This is due to the expected prognosis, i.e., a decreased manifestation of the disease, which also means correct administration of therapy. A relationship was found between the level of cfDNA and blood myeloperoxidase concentration in RA patients. The data obtained during the study suggest a possible connection between increased concentration of extracellular DNA, and activation of neutrophilic leukocytes in rheumatoid arthritis, with increased netosis in the affected joints.","PeriodicalId":21524,"journal":{"name":"Russian Journal of Immunology","volume":"22 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80198985","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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