Pub Date : 2023-06-01DOI: 10.15789/1563-0625-pao-2773
I. Maslennikova, I. Nekrasova, M. V. Kuznetsova
Recurrent urinary tract infections (UTIs) are associated primarily with the ability of Escherichia coli to form biofilms. The interaction of neutrophils, factors of innate immunity, with microorganisms in biofilms is difficult compared to planktonic forms due to the lack of direct contact, as well as due to the antiphagocytic action of the extracellular matrix of biofilms. The purpose of this study was evaluation of neutrophils phagocytic and oxidative activity during interaction with biofilms of uropathogenic E. coli (UPEC) DL82 and R44. Peripheral blood neutrophils from healthy men were isolated using ficoll-urographin double gradient, incubated for 1 h with bacterial cells from biofilms or their supernatants, then leukocytes functional activity was evaluated. Phagocytic activity of neutrophils was determined by the degree of bioluminescence inhibition of bioluminescent strain E. coli K12 TG1 lux+ (pXen) upon their absorption by neutrophils. Production of extracellular reactive oxygen species (ROS) was analyzed by the intensity of luminol-dependent chemiluminescence in spontaneous and stimulated by E. coli K12 variants. Significance of differences was determined using Student’s t-test at p < 0.05. It was found that neutrophils interaction with UPEC biofilm cells or supernatants did not affect the phagocytic activity. E. coli DL82 supernatants reduce neutrophils spontaneous ROS production compared to control and biofilm cells. E. coli R44 supernatants with a low virulence potential did not affect ROS production, while biofilm cells stimulated it. When assessing stimulated ROS production, exposure to R44 strain supernatants did not cause a decrease in neutrophils activation in response to an external stimulus (E. coli K12 cells). Preliminary contact of neutrophils with E. coli R44 bacteria resulted in a high and prolonged level of ROS production compared to the control. Neutrophils interaction with DL82 cells resulted in a higher level of ROS compared to supernatants, however a subsequent rapid depletion of neutrophils oxidative potential was observed. Thus, cells and supernatants of UPEC biofilms can determine the activation of neutrophils.
{"title":"Phagocytosis and oxidative activity of neutrophils after interaction with uropathogenic Escherichia coli biofilms","authors":"I. Maslennikova, I. Nekrasova, M. V. Kuznetsova","doi":"10.15789/1563-0625-pao-2773","DOIUrl":"https://doi.org/10.15789/1563-0625-pao-2773","url":null,"abstract":"Recurrent urinary tract infections (UTIs) are associated primarily with the ability of Escherichia coli to form biofilms. The interaction of neutrophils, factors of innate immunity, with microorganisms in biofilms is difficult compared to planktonic forms due to the lack of direct contact, as well as due to the antiphagocytic action of the extracellular matrix of biofilms. The purpose of this study was evaluation of neutrophils phagocytic and oxidative activity during interaction with biofilms of uropathogenic E. coli (UPEC) DL82 and R44. Peripheral blood neutrophils from healthy men were isolated using ficoll-urographin double gradient, incubated for 1 h with bacterial cells from biofilms or their supernatants, then leukocytes functional activity was evaluated. Phagocytic activity of neutrophils was determined by the degree of bioluminescence inhibition of bioluminescent strain E. coli K12 TG1 lux+ (pXen) upon their absorption by neutrophils. Production of extracellular reactive oxygen species (ROS) was analyzed by the intensity of luminol-dependent chemiluminescence in spontaneous and stimulated by E. coli K12 variants. Significance of differences was determined using Student’s t-test at p < 0.05. It was found that neutrophils interaction with UPEC biofilm cells or supernatants did not affect the phagocytic activity. E. coli DL82 supernatants reduce neutrophils spontaneous ROS production compared to control and biofilm cells. E. coli R44 supernatants with a low virulence potential did not affect ROS production, while biofilm cells stimulated it. When assessing stimulated ROS production, exposure to R44 strain supernatants did not cause a decrease in neutrophils activation in response to an external stimulus (E. coli K12 cells). Preliminary contact of neutrophils with E. coli R44 bacteria resulted in a high and prolonged level of ROS production compared to the control. Neutrophils interaction with DL82 cells resulted in a higher level of ROS compared to supernatants, however a subsequent rapid depletion of neutrophils oxidative potential was observed. Thus, cells and supernatants of UPEC biofilms can determine the activation of neutrophils.","PeriodicalId":37835,"journal":{"name":"Medical Immunology (Russia)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75199904","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 : 2023-06-01DOI: 10.15789/1563-0625-git-2778
E. Orlova, O. Loginova, O. Gorbunova, S. Shirshev
Galectin-9 is a b-galactoside binding lectin with expressed immunoregulatory activity. During pregnancy galectin-9 is produced by trophoblast cells and regulates the function of natural killer (NK) cells at the maternal-fetal interface via binding to Tim-3 (T-cell Ig and mucin domain-containing protein 3) molecules. Natural killer (NK) lymphocytes belong to the innate lymphoid cells, which have a cytotoxic effect on target cells and are capable of producing a large number of regulatory factors (cytokines, chemokines). Decidual NK have a tolerant phenotype and play a leading role in the regulation of invasive trophoblast growth and provide peripheral immune tolerance in the area of uteroplacental contact. Peripheral NK cells express Tim-3 molecules. Galectin-9 concentration is increased in peripheral blood during physiologic pregnancy. At pregnancy phenotype and functions of peripheral NK cells are changed to maintain the maternal–fetal immune tolerance. Peripheral NK cells migrate to the maternal-fetal interface and are transformed into a decidual NK-like phenotype cells. Galectin-9 concentration is decreased in women with a complicated pregnancy and miscarriage. However the galectin-9 effects on different NK cell subpopulations of peripheral blood are not investigated. Therefore, we studied the galectin-9 influence on phenotype transformation and Tim-3 expression of NK cells isolated from peripheral blood of healthy non-pregnant fertile women. CD56+NK cells were obtained by immunomagnetic separation and cultivated in vitro during 72 hours with cytokines (IL-2 and IL-15). Galectin-9 (5 ng/mL) and anti-Tim-3 (10 mg) antibodies were added to the NK cultures. Galectin-9 concentration is corresponded to its level during first trimester of physiologic pregnancy. The number of regulatory NK (CD16-CD56bright), cytotoxic NK (CD16+CD56dim/-) cells and Tim-3 expression on different NK subpopulations were assessed by flow cytometry. It was found that Tim-3 was expressed on all subpopulations of peripheral blood NK cells (CD16-CD56brightNK, CD16+CD56dimNK, CD16+CD56-NK). Incubation with galectin-9 increased the expression of Tim-3 on regulatory CD16-CD56brightNK cells and did not change on cytotoxic CD16+CD56dim/-NK cells. Galectin-9 reduced the percentage of cytotoxic CD16+CD56dimNK in culture, but did not influence the number of regulatory CD16-CD56bright NK and cytotoxic CD16+CD56-NK cells. Thus, galectin-9 regulates Tim-3 molecule expression and NK cell subpopulation distributions in vitro culture.
{"title":"Galectin-9 influences the Tim-3 molecule expression in natural killer different subpopulations","authors":"E. Orlova, O. Loginova, O. Gorbunova, S. Shirshev","doi":"10.15789/1563-0625-git-2778","DOIUrl":"https://doi.org/10.15789/1563-0625-git-2778","url":null,"abstract":"Galectin-9 is a b-galactoside binding lectin with expressed immunoregulatory activity. During pregnancy galectin-9 is produced by trophoblast cells and regulates the function of natural killer (NK) cells at the maternal-fetal interface via binding to Tim-3 (T-cell Ig and mucin domain-containing protein 3) molecules. Natural killer (NK) lymphocytes belong to the innate lymphoid cells, which have a cytotoxic effect on target cells and are capable of producing a large number of regulatory factors (cytokines, chemokines). Decidual NK have a tolerant phenotype and play a leading role in the regulation of invasive trophoblast growth and provide peripheral immune tolerance in the area of uteroplacental contact. Peripheral NK cells express Tim-3 molecules. Galectin-9 concentration is increased in peripheral blood during physiologic pregnancy. At pregnancy phenotype and functions of peripheral NK cells are changed to maintain the maternal–fetal immune tolerance. Peripheral NK cells migrate to the maternal-fetal interface and are transformed into a decidual NK-like phenotype cells. Galectin-9 concentration is decreased in women with a complicated pregnancy and miscarriage. However the galectin-9 effects on different NK cell subpopulations of peripheral blood are not investigated. Therefore, we studied the galectin-9 influence on phenotype transformation and Tim-3 expression of NK cells isolated from peripheral blood of healthy non-pregnant fertile women. CD56+NK cells were obtained by immunomagnetic separation and cultivated in vitro during 72 hours with cytokines (IL-2 and IL-15). Galectin-9 (5 ng/mL) and anti-Tim-3 (10 mg) antibodies were added to the NK cultures. Galectin-9 concentration is corresponded to its level during first trimester of physiologic pregnancy. The number of regulatory NK (CD16-CD56bright), cytotoxic NK (CD16+CD56dim/-) cells and Tim-3 expression on different NK subpopulations were assessed by flow cytometry. It was found that Tim-3 was expressed on all subpopulations of peripheral blood NK cells (CD16-CD56brightNK, CD16+CD56dimNK, CD16+CD56-NK). Incubation with galectin-9 increased the expression of Tim-3 on regulatory CD16-CD56brightNK cells and did not change on cytotoxic CD16+CD56dim/-NK cells. Galectin-9 reduced the percentage of cytotoxic CD16+CD56dimNK in culture, but did not influence the number of regulatory CD16-CD56bright NK and cytotoxic CD16+CD56-NK cells. Thus, galectin-9 regulates Tim-3 molecule expression and NK cell subpopulation distributions in vitro culture.","PeriodicalId":37835,"journal":{"name":"Medical Immunology (Russia)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72911376","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 : 2023-06-01DOI: 10.15789/1563-0625-apo-2678
I. Myagdieva, T. Abakumova, D. Dolgova, O. Gorshkov, T. Gening
The role of neutrophils in kidney cancer is currently being studied. Their role in carcinogenesis is ambiguous. As one of the most abundant blood leukocytes, neutrophils play an important role in cancer progression through multiple mechanisms, including promotion of angiogenesis, immunosuppression, and cancer metastasis. Neutrophils synthesize and release pro-angiogenic factors that are able to directly or indirectly stimulate the growth and migration of endothelial cells, which in turn causes the formation of new blood vessels from pre-existing ones. The production of various factors by neutrophils, including proangiogenic ones, is mediated by the expression of the genes of these molecules. Functional heterogeneity is characterized by differences in neutrophil gene expression patterns. The aim of this study was to evaluate the angiogenic potential of circulating neutrophils in kidney cancer. The object of the study were blood neutrophils of patients with verified clear cell kidney cancer at stage I (T1N0M0G1, n = 28, median age 60), stage II (T2N0M0G2, n = 15, median age 61) and stage III (T3N0M0G2, n = 15, median age 63) before surgery. The control group consisted of apparently healthy donors (n = 15, median age 54). Serum levels of IL-8 and VEGF-A were assessed by enzyme immunoassay. Expression of the CXCL8 and VEGF-A genes in circulating neutrophils was determined by reverse transcription quantitative PCR. As a result of our study, an increase in the level of IL-8 and VEGF-A in the blood serum of patients with kidney cancer in all studied groups compared with the control group was revealed. We observed a direct correlation between serum levels of IL-8 and VEGF-A in patients with kidney cancer (r = 0.429; p = 0.016), which confirms the relationship of these angiogenic factors. A significant increase in CXCL8 gene expression by circulating neutrophils was found in patients on II (2.91, Q0.25-Q0.75: (1.296-4.99), p = 0.02) and III (1.93, Q0.25-Q0.75: (0.755-11.36, p = 0.014) stages of kidney cancer compared with the control group (1.50, Q0.25-Q0.75: (0.80-4.05)). However, VEGF-A gene expression by circulating neutrophils did not differ from those in the control group. Blood neutrophils in kidney cancer exercise their angiogenic potential through the production of IL-8.
中性粒细胞在肾癌中的作用目前正在研究中。它们在癌变中的作用尚不明确。作为最丰富的血液白细胞之一，中性粒细胞通过促进血管生成、免疫抑制和肿瘤转移等多种机制在肿瘤进展中发挥重要作用。中性粒细胞合成并释放促血管生成因子，这些因子能够直接或间接刺激内皮细胞的生长和迁移，从而使原有血管形成新血管。中性粒细胞产生各种因子，包括促血管生成因子，是由这些分子的基因表达介导的。功能异质性的特征是中性粒细胞基因表达模式的差异。本研究的目的是评估肾癌中循环中性粒细胞的血管生成潜能。研究对象为术前确诊的透明细胞肾癌I期(T1N0M0G1, n = 28，中位年龄60)、II期(T2N0M0G2, n = 15，中位年龄61)和III期(T3N0M0G2, n = 15，中位年龄63)患者的血液中性粒细胞。对照组由表面健康的供体组成(n = 15，中位年龄54)。采用酶免疫分析法检测血清IL-8和VEGF-A水平。通过反转录定量PCR检测循环中性粒细胞中CXCL8和VEGF-A基因的表达。我们的研究结果显示，与对照组相比，所有研究组肾癌患者血清中IL-8和VEGF-A水平均有所升高。我们观察到肾癌患者血清IL-8和VEGF-A水平直接相关(r = 0.429;P = 0.016)，证实了这些血管生成因子之间的关系。II期(2.91,Q0.25-Q0.75: (1.296-4.99)， p = 0.02)和III期(1.93,Q0.25-Q0.75: (0.755-11.36, p = 0.014)肾癌患者与对照组(1.50,Q0.25-Q0.75:(0.80-4.05))相比，CXCL8基因通过循环中性粒细胞表达显著增加。然而，循环中性粒细胞的VEGF-A基因表达与对照组没有差异。肾癌中的血液中性粒细胞通过产生IL-8来发挥其血管生成潜能。
{"title":"Angiogenic potential of circulating peripheral blood neutrophils in kidney cancer","authors":"I. Myagdieva, T. Abakumova, D. Dolgova, O. Gorshkov, T. Gening","doi":"10.15789/1563-0625-apo-2678","DOIUrl":"https://doi.org/10.15789/1563-0625-apo-2678","url":null,"abstract":"The role of neutrophils in kidney cancer is currently being studied. Their role in carcinogenesis is ambiguous. As one of the most abundant blood leukocytes, neutrophils play an important role in cancer progression through multiple mechanisms, including promotion of angiogenesis, immunosuppression, and cancer metastasis. Neutrophils synthesize and release pro-angiogenic factors that are able to directly or indirectly stimulate the growth and migration of endothelial cells, which in turn causes the formation of new blood vessels from pre-existing ones. The production of various factors by neutrophils, including proangiogenic ones, is mediated by the expression of the genes of these molecules. Functional heterogeneity is characterized by differences in neutrophil gene expression patterns. The aim of this study was to evaluate the angiogenic potential of circulating neutrophils in kidney cancer. The object of the study were blood neutrophils of patients with verified clear cell kidney cancer at stage I (T1N0M0G1, n = 28, median age 60), stage II (T2N0M0G2, n = 15, median age 61) and stage III (T3N0M0G2, n = 15, median age 63) before surgery. The control group consisted of apparently healthy donors (n = 15, median age 54). Serum levels of IL-8 and VEGF-A were assessed by enzyme immunoassay. Expression of the CXCL8 and VEGF-A genes in circulating neutrophils was determined by reverse transcription quantitative PCR. As a result of our study, an increase in the level of IL-8 and VEGF-A in the blood serum of patients with kidney cancer in all studied groups compared with the control group was revealed. We observed a direct correlation between serum levels of IL-8 and VEGF-A in patients with kidney cancer (r = 0.429; p = 0.016), which confirms the relationship of these angiogenic factors. A significant increase in CXCL8 gene expression by circulating neutrophils was found in patients on II (2.91, Q0.25-Q0.75: (1.296-4.99), p = 0.02) and III (1.93, Q0.25-Q0.75: (0.755-11.36, p = 0.014) stages of kidney cancer compared with the control group (1.50, Q0.25-Q0.75: (0.80-4.05)). However, VEGF-A gene expression by circulating neutrophils did not differ from those in the control group. Blood neutrophils in kidney cancer exercise their angiogenic potential through the production of IL-8.","PeriodicalId":37835,"journal":{"name":"Medical Immunology (Russia)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73906263","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 : 2023-06-01DOI: 10.15789/1563-0625-rok-2751
O. Gorbunova, S. Shirshev
Pregnancy is a phenomenon of natural semi-allogeneic transplantation, since the fetus is half alien due to the expression of paternal antigens. It was found that the hypothalamic hormone kisspeptin during pregnancy is produced by the syncytiotrophoblast of the placenta and participates in the formation of a new specific hormonal background. Several forms of the hormone circulate in the blood of pregnant women: kisspeptin-10, kisspeptin-14 and kisspeptin-54 (according to the number of amino acid residues in the hormone molecule), but the main active form is kisspeptin-54. The main mechanism for the formation of immune tolerance during pregnancy is the induction of the expression of the enzyme indolamine-2,3- dioxygenase (IDO) by antigen-presenting cells of peripheral blood, resulting in the catalysis of tryptophan (Trp) to kynurenins (KYN) blocking the activation and causing apoptosis of cytotoxic CD8+T lymphocytes in the zone of contact of maternal immune cells with placental-fetal complex antigens. In addition, during pregnancy, an important role is assigned to the process of apoptosis, since activated cells can be potentially dangerous for the developing fetus. Immunocompetent blood cells express a specific membrane receptor of kisspeptin (KISS-1R). Since kisspeptin-54 enters the systemic circulation only during pregnancy, the hormone has an effect on immune cells only during this period.The aim of this work was to evaluate the effect of kisspeptin-54 in concentrations comparable to its levelduring physiological pregnancy on IDO activity and apoptosis of peripheral blood lymphocytes.Peripheral blood mononuclear cells (PBMC) obtained from 10 healthy non-pregnant women of reproductive age (from 23 to 32 years) were used as the object of the study. Lymphocyte apoptosis was assessed in PBMC suspension by staining with annexin-V and propidium iodide. The determination of the number of cells in the early and late stages of apoptosis was carried out in the isolated gate of lymphocytes. IDO activity in PBMC was determined spectrophotometrically by changes in the concentration of KYN, the first stable metabolite of the Trp decay pathway.It was found that kisspeptin-54 at a concentration of 4.6 pM corresponding to the second trimester of pregnancy significantly enhances the activity of IDO, increases the number of cells in the early and late stages of apoptosis. Thus, kisspeptin-54 is an important mechanism for controlling these processes during pregnancy, aimed at protecting the semi-allogeneic fetus from adverse immune reactions of the mother and the favorable development of pregnancy.
{"title":"Regulation of kisspeptin-54 activity of indolamine-2,3-dioxygenase and apoptosis of peripheral blood lymphocytes","authors":"O. Gorbunova, S. Shirshev","doi":"10.15789/1563-0625-rok-2751","DOIUrl":"https://doi.org/10.15789/1563-0625-rok-2751","url":null,"abstract":"Pregnancy is a phenomenon of natural semi-allogeneic transplantation, since the fetus is half alien due to the expression of paternal antigens. It was found that the hypothalamic hormone kisspeptin during pregnancy is produced by the syncytiotrophoblast of the placenta and participates in the formation of a new specific hormonal background. Several forms of the hormone circulate in the blood of pregnant women: kisspeptin-10, kisspeptin-14 and kisspeptin-54 (according to the number of amino acid residues in the hormone molecule), but the main active form is kisspeptin-54. The main mechanism for the formation of immune tolerance during pregnancy is the induction of the expression of the enzyme indolamine-2,3- dioxygenase (IDO) by antigen-presenting cells of peripheral blood, resulting in the catalysis of tryptophan (Trp) to kynurenins (KYN) blocking the activation and causing apoptosis of cytotoxic CD8+T lymphocytes in the zone of contact of maternal immune cells with placental-fetal complex antigens. In addition, during pregnancy, an important role is assigned to the process of apoptosis, since activated cells can be potentially dangerous for the developing fetus. Immunocompetent blood cells express a specific membrane receptor of kisspeptin (KISS-1R). Since kisspeptin-54 enters the systemic circulation only during pregnancy, the hormone has an effect on immune cells only during this period.The aim of this work was to evaluate the effect of kisspeptin-54 in concentrations comparable to its levelduring physiological pregnancy on IDO activity and apoptosis of peripheral blood lymphocytes.Peripheral blood mononuclear cells (PBMC) obtained from 10 healthy non-pregnant women of reproductive age (from 23 to 32 years) were used as the object of the study. Lymphocyte apoptosis was assessed in PBMC suspension by staining with annexin-V and propidium iodide. The determination of the number of cells in the early and late stages of apoptosis was carried out in the isolated gate of lymphocytes. IDO activity in PBMC was determined spectrophotometrically by changes in the concentration of KYN, the first stable metabolite of the Trp decay pathway.It was found that kisspeptin-54 at a concentration of 4.6 pM corresponding to the second trimester of pregnancy significantly enhances the activity of IDO, increases the number of cells in the early and late stages of apoptosis. Thus, kisspeptin-54 is an important mechanism for controlling these processes during pregnancy, aimed at protecting the semi-allogeneic fetus from adverse immune reactions of the mother and the favorable development of pregnancy.","PeriodicalId":37835,"journal":{"name":"Medical Immunology (Russia)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79338198","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 : 2023-06-01DOI: 10.15789/1563-0625-cat-2792
E. Orlova, O. Loginova
Monocytes play an important role in the systemic immune defense against pathogens and maintaining physiological pregnancy. During pregnancy peripheral monocytes migrate into the decidua and form the pool of decidual macrophages which participate in the formation and development of placental tissues. The population of peripheral blood monocytes is phenotypically and functionally heterogeneous. In humans, there are different monocyte subsets depending on the expression of CD14 and CD16. CD56-positive monocytes are found in healthy women. Their number is positively correlated with body mass index, body fat. Tim-3 (T cell Ig and mucin domain-containing protein 3) expression is observed in peripheral monocytes during pregnancy. It is known that peripheral monocyte functions effectively change at pregnancy to form the immune tolerance at the maternal-fetal interface and the systemic immune defense against pathogens. However, the monocyte phenotype shift during pregnancy remain poorly understood. Therefore, the aim of the study was to evaluate the CD56 and Tim-3 expressions in monocyte subsets in human pregnancy. Peripheral blood mononuclear cells were isolated from peripheral blood of pregnant women (gestational age 29 weeks (28-31) by density gradient centrifugation and analyzed by flow cytometry. Peripheral blood of healthy non-pregnant fertile women (in follicular phase of the menstrual cycle) aged 21-29 years was studied as control. Pregnant women had a lower percentage of classical CD14hi/CD16- monocytes in comparison with non-pregnant. The percentages of intermediate (CD14hi/CD16+) and non-classical (CD14low/CD16+) monocytes did not change. The CD56 molecule expression was observed in all monocyte subsets in pregnant and non-pregnant women. Pregnant women had a higher percentage of CD56-positive classical (CD14hiCD16-) and non-classical (CD14lowCD16+) monocytes than non-pregnant. The percentage of CD56-positive intermediate (CD14hiCD16+) monocytes did not change. The percentages of double-positive CD56+Tim-3+ classical (CD14hiCD16-) and non-classical (CD14lowCD16+) monocytes were increased in pregnant women. The numbers of double-positive CD56+Tim-3+intermediate (CD14hiCD16+) monocytes did not change. Thus, the CD56 and Tim-3 expressions in different monocyte subsets were changed in human pregnancy.
{"title":"CD56 and Tim-3 molecule expression in different monocyte subsets in physiological pregnancy","authors":"E. Orlova, O. Loginova","doi":"10.15789/1563-0625-cat-2792","DOIUrl":"https://doi.org/10.15789/1563-0625-cat-2792","url":null,"abstract":"Monocytes play an important role in the systemic immune defense against pathogens and maintaining physiological pregnancy. During pregnancy peripheral monocytes migrate into the decidua and form the pool of decidual macrophages which participate in the formation and development of placental tissues. The population of peripheral blood monocytes is phenotypically and functionally heterogeneous. In humans, there are different monocyte subsets depending on the expression of CD14 and CD16. CD56-positive monocytes are found in healthy women. Their number is positively correlated with body mass index, body fat. Tim-3 (T cell Ig and mucin domain-containing protein 3) expression is observed in peripheral monocytes during pregnancy. It is known that peripheral monocyte functions effectively change at pregnancy to form the immune tolerance at the maternal-fetal interface and the systemic immune defense against pathogens. However, the monocyte phenotype shift during pregnancy remain poorly understood. Therefore, the aim of the study was to evaluate the CD56 and Tim-3 expressions in monocyte subsets in human pregnancy. Peripheral blood mononuclear cells were isolated from peripheral blood of pregnant women (gestational age 29 weeks (28-31) by density gradient centrifugation and analyzed by flow cytometry. Peripheral blood of healthy non-pregnant fertile women (in follicular phase of the menstrual cycle) aged 21-29 years was studied as control. Pregnant women had a lower percentage of classical CD14hi/CD16- monocytes in comparison with non-pregnant. The percentages of intermediate (CD14hi/CD16+) and non-classical (CD14low/CD16+) monocytes did not change. The CD56 molecule expression was observed in all monocyte subsets in pregnant and non-pregnant women. Pregnant women had a higher percentage of CD56-positive classical (CD14hiCD16-) and non-classical (CD14lowCD16+) monocytes than non-pregnant. The percentage of CD56-positive intermediate (CD14hiCD16+) monocytes did not change. The percentages of double-positive CD56+Tim-3+ classical (CD14hiCD16-) and non-classical (CD14lowCD16+) monocytes were increased in pregnant women. The numbers of double-positive CD56+Tim-3+intermediate (CD14hiCD16+) monocytes did not change. Thus, the CD56 and Tim-3 expressions in different monocyte subsets were changed in human pregnancy.","PeriodicalId":37835,"journal":{"name":"Medical Immunology (Russia)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81662666","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 : 2023-06-01DOI: 10.15789/1563-0625-svp-2735
Y. Filippova, A. Alekseeva, E. V. Devyatova, K. A. Rusakova, A. Burmistrova
The autism spectrum disorders (ASD) are now widely accepted as a pervasive, complex, heterogeneous neurodevelopmental disorders with multiple etiologies, subtypes, and developmental trajectories. There are no available and effective biomarkers for them. Immune dysfunction is seen as an important risk factor contributing to the neurodevelopmental deficit in ASD, and is signified, among other things, by an imbalance of cytokines in the brain and on the periphery. In recent years, saliva has been proposed as a biological material for diagnosing ASD, due to the accessibility and non-invasiveness of the method for its production. However, the question of whether salivary cytokine levels may be used as effective early biomarkers for autism requires further research, including saliva versus plasma/serum comparisons.Aim: a comparative analysis of the levels of cytokines: IL-6, IFNγ, TNFα, IL-1β, IL-4, IL-10, in saliva and blood plasma to identify possible markers of ASD and their severity in children.The study included 11 children with typical neurodevelopment (TDC) and 55 children with ASD, among whom 37 children had mild or moderate autism (according to CARS), and 18 children had severe autism. Samples of unstimulated mixed saliva and venous blood were simultaneously collected from all children. Salivary concentrations of cytokines: IL-6, IFNγ, TNFα, IL-1β, IL-4, IL-10 were determined by multiplex Luminex™ analysis. Plasma levels of cytokines were assessed by ELISA. Differences between groups were tested using the Kruskal-Wallis U-test with post-hoc Conover-Inman comparisons, between samples (saliva/ plasma) are using the Wilcoxon signed-rank test. The correlation between the concentrations of cytokines in plasma and saliva was determined using linear regression by the RMA method.In all examined groups, the levels of IL-6, IFNγ and IL-10 in saliva were significantly lower, and TNFα, IL-1β and IL-4 were higher than the corresponding levels of the same cytokines in plasma. Regardless of health/ disease status, no significant correlations were found between salivary and plasma cytokine levels in children. IL-1β levels were significantly lower and IL-10 levels were higher in the saliva of both groups of children with ASD compared with TDC. No significant differences in salivary cytokine concentrations were found between children with mild and severe ASD.Thus, salivary cytokines can be used as markers of ASD in children, but not the severity of the condition. The absence of correlations in the levels of some pro/anti-inflammatory cytokines between saliva and blood plasma may probably indicate a special immunological status of an ecological niche, the oral cavity.
{"title":"Saliva versus plasma cytokines as possible predictors of autism severity","authors":"Y. Filippova, A. Alekseeva, E. V. Devyatova, K. A. Rusakova, A. Burmistrova","doi":"10.15789/1563-0625-svp-2735","DOIUrl":"https://doi.org/10.15789/1563-0625-svp-2735","url":null,"abstract":"The autism spectrum disorders (ASD) are now widely accepted as a pervasive, complex, heterogeneous neurodevelopmental disorders with multiple etiologies, subtypes, and developmental trajectories. There are no available and effective biomarkers for them. Immune dysfunction is seen as an important risk factor contributing to the neurodevelopmental deficit in ASD, and is signified, among other things, by an imbalance of cytokines in the brain and on the periphery. In recent years, saliva has been proposed as a biological material for diagnosing ASD, due to the accessibility and non-invasiveness of the method for its production. However, the question of whether salivary cytokine levels may be used as effective early biomarkers for autism requires further research, including saliva versus plasma/serum comparisons.Aim: a comparative analysis of the levels of cytokines: IL-6, IFNγ, TNFα, IL-1β, IL-4, IL-10, in saliva and blood plasma to identify possible markers of ASD and their severity in children.The study included 11 children with typical neurodevelopment (TDC) and 55 children with ASD, among whom 37 children had mild or moderate autism (according to CARS), and 18 children had severe autism. Samples of unstimulated mixed saliva and venous blood were simultaneously collected from all children. Salivary concentrations of cytokines: IL-6, IFNγ, TNFα, IL-1β, IL-4, IL-10 were determined by multiplex Luminex™ analysis. Plasma levels of cytokines were assessed by ELISA. Differences between groups were tested using the Kruskal-Wallis U-test with post-hoc Conover-Inman comparisons, between samples (saliva/ plasma) are using the Wilcoxon signed-rank test. The correlation between the concentrations of cytokines in plasma and saliva was determined using linear regression by the RMA method.In all examined groups, the levels of IL-6, IFNγ and IL-10 in saliva were significantly lower, and TNFα, IL-1β and IL-4 were higher than the corresponding levels of the same cytokines in plasma. Regardless of health/ disease status, no significant correlations were found between salivary and plasma cytokine levels in children. IL-1β levels were significantly lower and IL-10 levels were higher in the saliva of both groups of children with ASD compared with TDC. No significant differences in salivary cytokine concentrations were found between children with mild and severe ASD.Thus, salivary cytokines can be used as markers of ASD in children, but not the severity of the condition. The absence of correlations in the levels of some pro/anti-inflammatory cytokines between saliva and blood plasma may probably indicate a special immunological status of an ecological niche, the oral cavity.","PeriodicalId":37835,"journal":{"name":"Medical Immunology (Russia)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85073962","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 : 2023-06-01DOI: 10.15789/1563-0625-ibc-2810
G. Jiemuratova
Hematopoiesis is a complex process that requires a specific set of blood components to function properly. Blood diseases can result from imbalances or deficiencies in these components. The body has physiological sensors that respond to environmental changes by maintaining elemental homeostasis. A deficiency in one micronutrient can lead to imbalances in others. The purpose of this study was to investigate the role and interaction of copper, cobalt, and iron in hematopoiesis and to determine the prevalence of anemia in children living in the Aral Sea region.A total of 1120 children and adolescents were examined, and their physical development was measured using anthropometric measurements and laboratory tests. Hair samples were analyzed to determine the children's micronutrient status. The results revealed that 78% of the children had a decrease in hemoglobin, and anemia was more prevalent in adolescents. A correlation was found between high growth and increased levels of erythrocytes and hemoglobin. The study also identified the most common hypomicroelementoses in the Aral Sea region, including copper deficiency in 98.4% of cases, cobalt deficiency in 92.1%, and zinc deficiency in 57.8%.The study also analyzed the ratio of trace elements, revealing an increased Fe/Cu and Fe/Cu ratio in all age groups. Imbalances and deficiencies in copper, cobalt, zinc, and manganese were found to contribute to the development of anemia in children. Hair analysis for trace elements was shown to be significant in the differential diagnosis and treatment of children with anemia.In conclusion, the study highlights the importance of maintaining a proper balance of trace elements in hematopoiesis. Deficiencies in copper, cobalt, zinc, and manganese can contribute to anemia in children, and hair analysis can be used to diagnose and treat the condition. Further research is needed to better understand the role of trace elements in hematopoiesis and their impact on human health.
{"title":"Interplay between copper and cobalt in hematopoiesis and the impact of their deficiency on anemia development","authors":"G. Jiemuratova","doi":"10.15789/1563-0625-ibc-2810","DOIUrl":"https://doi.org/10.15789/1563-0625-ibc-2810","url":null,"abstract":"Hematopoiesis is a complex process that requires a specific set of blood components to function properly. Blood diseases can result from imbalances or deficiencies in these components. The body has physiological sensors that respond to environmental changes by maintaining elemental homeostasis. A deficiency in one micronutrient can lead to imbalances in others. The purpose of this study was to investigate the role and interaction of copper, cobalt, and iron in hematopoiesis and to determine the prevalence of anemia in children living in the Aral Sea region.A total of 1120 children and adolescents were examined, and their physical development was measured using anthropometric measurements and laboratory tests. Hair samples were analyzed to determine the children's micronutrient status. The results revealed that 78% of the children had a decrease in hemoglobin, and anemia was more prevalent in adolescents. A correlation was found between high growth and increased levels of erythrocytes and hemoglobin. The study also identified the most common hypomicroelementoses in the Aral Sea region, including copper deficiency in 98.4% of cases, cobalt deficiency in 92.1%, and zinc deficiency in 57.8%.The study also analyzed the ratio of trace elements, revealing an increased Fe/Cu and Fe/Cu ratio in all age groups. Imbalances and deficiencies in copper, cobalt, zinc, and manganese were found to contribute to the development of anemia in children. Hair analysis for trace elements was shown to be significant in the differential diagnosis and treatment of children with anemia.In conclusion, the study highlights the importance of maintaining a proper balance of trace elements in hematopoiesis. Deficiencies in copper, cobalt, zinc, and manganese can contribute to anemia in children, and hair analysis can be used to diagnose and treat the condition. Further research is needed to better understand the role of trace elements in hematopoiesis and their impact on human health.","PeriodicalId":37835,"journal":{"name":"Medical Immunology (Russia)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80212790","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 : 2023-06-01DOI: 10.15789/1563-0625-feo-2734
V. V. Vlasova, L. Korolevskaya, O. Loginova, N. Shmagel, E. V. Saidakova
Infection with hepatitis C virus (HCV) is common among HIV-positive patients, with up to 50% of them being coinfected in Russia. While highly active antiretroviral therapy (HAART) suppresses HIV replication and restores the immune system of HIV-infected subjects, HCV coinfection interferes with CD4+T cell regeneration and increases the risk of patients’ morbidity and mortality. During HAART, HIVinfection progression and the immune system restoration efficiency largely depend on immune activation and CD4+T cell exhaustion. This study determined the level of activation, exhaustion, and cytokine production in CD4+T cells obtained from the peripheral blood of HAART-treated HIV/HCV coinfected and HIV monoinfected subjects. The study comprised 11 HIV/HCV coinfected individuals and 10 HIV monoinfected patients receiving HAART for more than two years, with a control group of 10 volunteers without the signs of HIV or HCV infections. Compared with healthy controls, HIV/HCV coinfected patients had an increased frequency of activated CD38+HLA-DR+ CD4+T lymphocytes (p < 0.05), a higher level of CD4+T cell exhaustion determined according to the TIGIT expression density per cell (p < 0.05), and a greater proportion of interferon-gamma (IFNγ)-producing CD4+T lymphocytes following activation (p < 0.05). The frequency of IFNγ-producing CD4+T cells in the donors’ blood positively correlated with the proportion of activated CD4+T cells (R = 0.514, p < 0.01). Despite having a large number of IFNγ-producing CD4+T lymphocytes, the HIV/HCV coinfected patients’ average production of IFNγ by CD4+T cells was significantly lower than that in healthy controls (p < 0.05). The IFNγ production in CD4+T lymphocytes did not depend on activation (p > 0.05). However, a negative correlation was established between the IFNγ production and the level of CD4+T cell exhaustion (R = -0.400, p < 0.05). The letter was also found to inversely correlate with the CD4+T cell counts in the donors’ peripheral blood (R = -0.598, p < 0.01). These data suggest that HCV coinfection leads to pronounced functional exhaustion of CD4+T cells and may aggravate the course of HIVinfection in patients receiving HAART.
丙型肝炎病毒(HCV)感染在艾滋病毒阳性患者中很常见，在俄罗斯，多达50%的患者合并感染。虽然高活性抗逆转录病毒疗法(HAART)抑制HIV复制并恢复HIV感染者的免疫系统，但HCV合并感染干扰CD4+T细胞再生并增加患者发病率和死亡率的风险。在HAART治疗期间，hiv感染的进展和免疫系统的恢复效率在很大程度上取决于免疫激活和CD4+T细胞衰竭。本研究测定了经haart治疗的HIV/HCV合并感染者和HIV单感染者外周血中CD4+T细胞的激活、衰竭和细胞因子产生水平。该研究包括11名HIV/HCV合并感染者和10名接受HAART治疗超过两年的HIV单感染者，对照组为10名没有HIV或HCV感染迹象的志愿者。与健康对照组相比，HIV/HCV共感染患者激活CD38+HLA-DR+ CD4+T淋巴细胞的频率增加(p < 0.05)，根据每个细胞的TIGIT表达密度确定的CD4+T细胞耗竭水平更高(p < 0.05)，激活后产生干扰素γ (IFNγ)的CD4+T淋巴细胞比例更高(p < 0.05)。供者血液中产生ifn γ的CD4+T细胞频率与CD4+T细胞活化比例呈正相关(R = 0.514, p < 0.01)。HIV/HCV共感染患者虽然有大量产生IFNγ的CD4+T淋巴细胞，但CD4+T细胞平均产生IFNγ的水平明显低于健康对照组(p < 0.05)。CD4+T淋巴细胞中IFNγ的产生不依赖于活化(p > 0.05)。然而，IFNγ的产生与CD4+T细胞衰竭水平呈负相关(R = -0.400, p < 0.05)。字母与献血者外周血CD4+T细胞计数呈负相关(R = -0.598, p < 0.01)。这些数据表明，HCV合并感染导致CD4+T细胞明显的功能衰竭，并可能加重HAART患者的hiv感染过程。
{"title":"Functional exhaustion of CD4+T cells in HIV/HCV coinfected HAART-treated patients","authors":"V. V. Vlasova, L. Korolevskaya, O. Loginova, N. Shmagel, E. V. Saidakova","doi":"10.15789/1563-0625-feo-2734","DOIUrl":"https://doi.org/10.15789/1563-0625-feo-2734","url":null,"abstract":"Infection with hepatitis C virus (HCV) is common among HIV-positive patients, with up to 50% of them being coinfected in Russia. While highly active antiretroviral therapy (HAART) suppresses HIV replication and restores the immune system of HIV-infected subjects, HCV coinfection interferes with CD4+T cell regeneration and increases the risk of patients’ morbidity and mortality. During HAART, HIVinfection progression and the immune system restoration efficiency largely depend on immune activation and CD4+T cell exhaustion. This study determined the level of activation, exhaustion, and cytokine production in CD4+T cells obtained from the peripheral blood of HAART-treated HIV/HCV coinfected and HIV monoinfected subjects. The study comprised 11 HIV/HCV coinfected individuals and 10 HIV monoinfected patients receiving HAART for more than two years, with a control group of 10 volunteers without the signs of HIV or HCV infections. Compared with healthy controls, HIV/HCV coinfected patients had an increased frequency of activated CD38+HLA-DR+ CD4+T lymphocytes (p < 0.05), a higher level of CD4+T cell exhaustion determined according to the TIGIT expression density per cell (p < 0.05), and a greater proportion of interferon-gamma (IFNγ)-producing CD4+T lymphocytes following activation (p < 0.05). The frequency of IFNγ-producing CD4+T cells in the donors’ blood positively correlated with the proportion of activated CD4+T cells (R = 0.514, p < 0.01). Despite having a large number of IFNγ-producing CD4+T lymphocytes, the HIV/HCV coinfected patients’ average production of IFNγ by CD4+T cells was significantly lower than that in healthy controls (p < 0.05). The IFNγ production in CD4+T lymphocytes did not depend on activation (p > 0.05). However, a negative correlation was established between the IFNγ production and the level of CD4+T cell exhaustion (R = -0.400, p < 0.05). The letter was also found to inversely correlate with the CD4+T cell counts in the donors’ peripheral blood (R = -0.598, p < 0.01). These data suggest that HCV coinfection leads to pronounced functional exhaustion of CD4+T cells and may aggravate the course of HIVinfection in patients receiving HAART.","PeriodicalId":37835,"journal":{"name":"Medical Immunology (Russia)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87963232","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 : 2023-06-01DOI: 10.15789/1563-0625-doa-2824
E. Volosnikova, D. Shcherbakov, V. V. Ermolaev, N. V. Volkova, O. Kaplina, M. B. Borgoyakova, E. Danilenko
The use of modern subunit vaccines involves adjuvant introduction into their composition. Currently, the search for new and improvement of existing adjuvant systems is actively underway. Squalene- based adjuvants are well-known and approved in a number of countries for clinical use in influenza vaccines. Our study was devoted to the development of an adjuvant composition on the basis of squalene. The resulting adjuvants were composed in a form of oil emulsion containing a hydrophilic and hydrophobic phase. The stability of the emulsion was achieved by treating it with ultrasound at a frequency of 22 kHz. Particle sizes of the obtained emulsions were examined with the use of an electron microscope. The particle size was calculated to be 50-80 nm for the majority of particles (84%). Adjuvant activity was evaluated in 100 male Balb/C mice, weighing 16-18 g. To assess the humoral immune response, immunization was performed twice, with a 14-day interval, by intramuscular injection of 200 mL per animal. The receptor-binding domain (RBD) of the surface S protein of the severe acute respiratory syndrome coronavirus 2 (Delta variant (B.1.617.2)) or ovalbumin (OVA) from chicken eggs were used as antigens. RBD was administered at a dose of 50 mg/animal; OVA was administered at two doses (1 mg or 5 mg/animal). An antigen with aluminum hydroxide was used as a positive control; a saline solution was used as a negative control. The effectiveness of the obtained adjuvants was determined by measuring the titers of specific antibodies in mouse sera in ELISA assays using the recombinant RBD of SARS-CoV-2 S-protein or ovalbumin from chicken eggs. It was shown that the use of squalene-based adjuvants increased the antigens’ immunogenicity. The average titers of specific antibodies against RBD in the experimental group were 4 times higher than in the group immunized with RBD adjuvanted with aluminum hydroxide. An increase in immunogenicity of the antigen adjuvanted with squalene was also observed in the experimental OVA-group. Thus, it was shown that the developed squalene-based adjuvant compositions could be an alternative to the traditional adjuvants based on aluminum salts.
{"title":"Development of a vaccine adjuvant based on squalene and study of its adjuvant properties","authors":"E. Volosnikova, D. Shcherbakov, V. V. Ermolaev, N. V. Volkova, O. Kaplina, M. B. Borgoyakova, E. Danilenko","doi":"10.15789/1563-0625-doa-2824","DOIUrl":"https://doi.org/10.15789/1563-0625-doa-2824","url":null,"abstract":"The use of modern subunit vaccines involves adjuvant introduction into their composition. Currently, the search for new and improvement of existing adjuvant systems is actively underway. Squalene- based adjuvants are well-known and approved in a number of countries for clinical use in influenza vaccines. Our study was devoted to the development of an adjuvant composition on the basis of squalene. The resulting adjuvants were composed in a form of oil emulsion containing a hydrophilic and hydrophobic phase. The stability of the emulsion was achieved by treating it with ultrasound at a frequency of 22 kHz. Particle sizes of the obtained emulsions were examined with the use of an electron microscope. The particle size was calculated to be 50-80 nm for the majority of particles (84%). Adjuvant activity was evaluated in 100 male Balb/C mice, weighing 16-18 g. To assess the humoral immune response, immunization was performed twice, with a 14-day interval, by intramuscular injection of 200 mL per animal. The receptor-binding domain (RBD) of the surface S protein of the severe acute respiratory syndrome coronavirus 2 (Delta variant (B.1.617.2)) or ovalbumin (OVA) from chicken eggs were used as antigens. RBD was administered at a dose of 50 mg/animal; OVA was administered at two doses (1 mg or 5 mg/animal). An antigen with aluminum hydroxide was used as a positive control; a saline solution was used as a negative control. The effectiveness of the obtained adjuvants was determined by measuring the titers of specific antibodies in mouse sera in ELISA assays using the recombinant RBD of SARS-CoV-2 S-protein or ovalbumin from chicken eggs. It was shown that the use of squalene-based adjuvants increased the antigens’ immunogenicity. The average titers of specific antibodies against RBD in the experimental group were 4 times higher than in the group immunized with RBD adjuvanted with aluminum hydroxide. An increase in immunogenicity of the antigen adjuvanted with squalene was also observed in the experimental OVA-group. Thus, it was shown that the developed squalene-based adjuvant compositions could be an alternative to the traditional adjuvants based on aluminum salts.","PeriodicalId":37835,"journal":{"name":"Medical Immunology (Russia)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89145369","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 : 2023-06-01DOI: 10.15789/1563-0625-scp-2694
N. Lazareva, I. Kudryavtsev, O. Baranova, D. Isakov, M. Serebriakova, A. A. Bazhanov, N. A. Arsentieva, N. Liubimova, T. Ses’, M. Ilkovich, A. Totolian
Immune cell hyperactivation along with cytokines they overproduce plays an important role in sarcoidosis and related disease pathogenesis. A central place in the immunopathogenesis of sarcoidosis is held by diverse cell-mediated reactions governed by T helper (Th) cell populations including Th17 subsets and relevant signature cytokines. We studied peripheral blood plasma samples of the patients with sarcoidosis (n = 123): 18% with acute and 82% with chronic course. The control group — samples from healthy volunteers (n = 43). T cell subset composition was assessed by flow cytometry. Cytokine concentrations (pg/mL) were measured by multiplex analysis using xMAP technology (Luminex). The level of “classical” Th17 turned out to be significantly reduced in acute vs chronic sarcoidosis: 28.3% vs 33.3% (p = 0.046). The level of “double-positive” Th17 (DP Th17) was significantly increased in chronic and acute vs control group: 31.7% and 34.2% vs 26.2% (p < 0.001 in both cases), without differences patient inter-group; “non-classical” Th17.1 were shown to have significantly reduced level only in chronic vs healthy subjects: 27.9% and 35.9% (p < 0.001). Clinical and laboratory diagnostic characteristics for blood DP Th17 levels in CD45RA-negative Th effector memory cells in sarcoidosis: in acute sarcoidosis vs healthy subjects, they were characterized by sensitivity — 82%; specificity — 71%, whereas in chronic: 67% and 56%, respectively. In patients with sarcoidosis vs healthy subjects were found to have significantly increased level of IL-12 (p70) — 1.3 vs 0.56, p = 0.028; IL-17A — 1.5 vs 0.43, p < 0.001; IFNγ — 4.1 vs 1.1, p < 0.001; TNFα — 21.7 vs 6.7, p < 0.001. Thus, CCR6+ Th17 and DP Th17 subsets and relevant signature cytokines are important in diagnostics of sarcoidosis of varying clinical course: a direct correlation was shown between the level of angiotensin-converting enzyme activity and percentage of memory DP Th17; disease progression vs regression had significantly reduced absolute number of total CD45RA- memory and CM Th17; extrapulmonary manifestations had a significantly increased percentage of DP Th17 CD45RA- and EM DP Th17; in chronic sarcoidosis are significantly increased concentration of IL-17A, IFNγ, IL-12 and positively correlation between IFNγ and the activity of angiotensin-converting enzyme.
免疫细胞的过度活化及其产生的细胞因子在结节病及其相关疾病的发病机制中起重要作用。结节病的免疫发病机制是由辅助性T细胞群(包括Th17亚群和相关的特征细胞因子)控制的多种细胞介导的反应所控制的。我们研究了123例结节病患者的外周血血浆样本:18%为急性病程，82%为慢性病程。对照组——健康志愿者样本(n = 43)。流式细胞术检测T细胞亚群组成。细胞因子浓度(pg/mL)采用多重分析xMAP技术(Luminex)测定。“经典”Th17水平在急性结节病和慢性结节病中显著降低:28.3%比33.3% (p = 0.046)。“双阳性”Th17 (DP Th17)水平在慢性和急性组与对照组相比显著升高:分别为31.7%和34.2%对26.2% (p < 0.001)，组间无差异;“非经典”Th17.1水平仅在慢性受试者和健康受试者中显著降低:27.9%和35.9% (p < 0.001)。结节病患者cd45ra阴性Th效应记忆细胞血液中DP Th17水平的临床和实验室诊断特征:急性结节病患者与健康受试者的敏感性为82%;特异性为71%，而慢性:分别为67%和56%。结节病患者与健康人相比IL-12水平显著升高(p70)——1.3 vs 0.56, p = 0.028;IL-17A - 1.5 vs 0.43, p < 0.001;IFNγ - 4.1 vs 1.1, p < 0.001;TNFα - 21.7 vs 6.7, p < 0.001。因此，CCR6+ Th17和DP Th17亚群以及相关的特征细胞因子在不同临床病程的结节病诊断中很重要:血管紧张素转换酶活性水平与记忆DP Th17百分比之间存在直接相关性;疾病进展与消退显著降低了总CD45RA-记忆和CM Th17的绝对数量;肺外表现DP Th17、CD45RA-和EM DP Th17的比例显著增加;慢性结节病患者IL-17A、IFNγ、IL-12浓度显著升高，且IFNγ与血管紧张素转换酶活性呈正相关。
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