Pub Date : 2020-12-01DOI: 10.15789/1563-0625-trb-2032
O. Smirnova, V. Tsukanov, A. Sinyakov, O. Moskalenko, N. G. Elmanova, E. S. Ovcharenko
Chronic atrophic gastritis and gastric cancer represent distinct steps of one pathogenic process. The risk of developing cancer of the stomach is directly proportional to the degree of atrophic changes simultaneously detected in antral segment and in the body of the stomach. The role of immune system in transformation of precancerous diseases into cancer is beyond doubt. During development of the malignant disease, the changes in lipid peroxidation systems – antioxidant defense become significant and contribute to the progression of the tumor and the development of metastases. A simultaneous study of lipid peroxidation and antioxidant defense indices along with phagocytic activity will allow us to evaluate relative contribution of these processes to development of chronic atrophic gastritis and gastric cancer. Purpose of the present study was to assess correlations between the lipid peroxidation indices, i.e. antioxidant protection, and chemiluminescent activity of neutrophilic granulocytes and monocytes in chronic atrophic gastritis and gastric cancer. Forty patients with chronic gastritis, 22 patients with chronic atrophic gastritis and 40 patients with gastric cancer were examined. The control group consisted of 50 practically healthy age-matched volunteers. Evaluation of spontaneous and induced production of reactive oxygen species by neutrophils and monocytes was carried out by chemiluminescent analysis. The parameters of lipid peroxidation/ antioxidant protection were determined by spectrophotometric methods. Statistical data processing was carried out using the Statistica v. 8.0 program (StatSoft Inc., USA). The normal distribution of indices was tested using the Kolmogorov–Smirnov method (adjusted by Lillefors). Quantitative indicators, given the normal distribution, were described using the median (Me) and interquartile scatter (Q0.25-Q0.75). To study statistical significance of differences between quantitative characteristics, the Mann–Whitney test was used. To study strength of relationships of these indicators, the Pearson rank correlation coefficient (r) was calculated. The critical significance level (p) when testing statistical hypotheses was taken equal to 0.05. Correlation analysis showed that the weight of positive correlations increases in patients with chronic atrophic gastritis, and it decreases in patients with gastric cancer, the strength of the correlation dependence and new relationships appear between chemiluminescent activity of neutrophils and monocytes in a spontaneous and induced state, and the amounts of malonic dialdehyde, enzyme activities of superoxide dismutase and catalase. In chronic atrophic gastritis and gastric cancer we have established the features of correlation patterns between lipid peroxidation/antioxidant protection indices, and activity of neutrophils and monocytes.
慢性萎缩性胃炎和胃癌是一个不同的致病过程。发生胃癌的风险与同时在胃窦段和胃体检测到的萎缩变化程度成正比。免疫系统在癌前病变转化为癌症中的作用是毋庸置疑的。在恶性疾病的发展过程中,脂质过氧化系统-抗氧化防御的变化变得显著,并有助于肿瘤的进展和转移的发展。同时研究脂质过氧化和抗氧化防御指数以及吞噬活性将使我们能够评估这些过程在慢性萎缩性胃炎和胃癌发展中的相对贡献。本研究旨在探讨慢性萎缩性胃炎和胃癌中脂质过氧化指标(抗氧化保护)与中性粒细胞和单核细胞化学发光活性的相关性。对40例慢性胃炎、22例慢性萎缩性胃炎和40例胃癌患者进行了检查。对照组由50名年龄相当的健康志愿者组成。通过化学发光分析评估中性粒细胞和单核细胞自发和诱导产生的活性氧。采用分光光度法测定脂质过氧化/抗氧化保护参数。统计数据处理使用Statistica v. 8.0程序(StatSoft Inc., USA)。各项指标的正态分布采用Kolmogorov-Smirnov方法进行检验(经Lillefors校正)。定量指标,鉴于正态分布,用中位数(Me)和四分位数散点(Q0.25-Q0.75)来描述。为研究数量特征间差异的统计学意义,采用Mann-Whitney检验。为了研究这些指标之间的关系强度,我们计算了Pearson秩相关系数(r)。检验统计假设时的临界显著性水平(p)为0.05。相关性分析显示,慢性萎缩性胃炎患者的正相关权值增加,胃癌患者的正相关权值降低,自发和诱导状态下中性粒细胞和单核细胞的化学发光活性与丙二醛含量、超氧化物歧化酶和过氧化氢酶活性之间的相关依赖强度增强,出现了新的关系。在慢性萎缩性胃炎和胃癌中,我们建立了脂质过氧化/抗氧化保护指数与中性粒细胞和单核细胞活性的相关模式特征。
{"title":"Typical relationships between the indexes of lipid peroxidation/ antioxidant protection and immune regulation in the patients with chronic atrophic gastritis and stomach cancer","authors":"O. Smirnova, V. Tsukanov, A. Sinyakov, O. Moskalenko, N. G. Elmanova, E. S. Ovcharenko","doi":"10.15789/1563-0625-trb-2032","DOIUrl":"https://doi.org/10.15789/1563-0625-trb-2032","url":null,"abstract":"Chronic atrophic gastritis and gastric cancer represent distinct steps of one pathogenic process. The risk of developing cancer of the stomach is directly proportional to the degree of atrophic changes simultaneously detected in antral segment and in the body of the stomach. The role of immune system in transformation of precancerous diseases into cancer is beyond doubt. During development of the malignant disease, the changes in lipid peroxidation systems – antioxidant defense become significant and contribute to the progression of the tumor and the development of metastases. A simultaneous study of lipid peroxidation and antioxidant defense indices along with phagocytic activity will allow us to evaluate relative contribution of these processes to development of chronic atrophic gastritis and gastric cancer. Purpose of the present study was to assess correlations between the lipid peroxidation indices, i.e. antioxidant protection, and chemiluminescent activity of neutrophilic granulocytes and monocytes in chronic atrophic gastritis and gastric cancer. Forty patients with chronic gastritis, 22 patients with chronic atrophic gastritis and 40 patients with gastric cancer were examined. The control group consisted of 50 practically healthy age-matched volunteers. Evaluation of spontaneous and induced production of reactive oxygen species by neutrophils and monocytes was carried out by chemiluminescent analysis. The parameters of lipid peroxidation/ antioxidant protection were determined by spectrophotometric methods. Statistical data processing was carried out using the Statistica v. 8.0 program (StatSoft Inc., USA). The normal distribution of indices was tested using the Kolmogorov–Smirnov method (adjusted by Lillefors). Quantitative indicators, given the normal distribution, were described using the median (Me) and interquartile scatter (Q0.25-Q0.75). To study statistical significance of differences between quantitative characteristics, the Mann–Whitney test was used. To study strength of relationships of these indicators, the Pearson rank correlation coefficient (r) was calculated. The critical significance level (p) when testing statistical hypotheses was taken equal to 0.05. Correlation analysis showed that the weight of positive correlations increases in patients with chronic atrophic gastritis, and it decreases in patients with gastric cancer, the strength of the correlation dependence and new relationships appear between chemiluminescent activity of neutrophils and monocytes in a spontaneous and induced state, and the amounts of malonic dialdehyde, enzyme activities of superoxide dismutase and catalase. In chronic atrophic gastritis and gastric cancer we have established the features of correlation patterns between lipid peroxidation/antioxidant protection indices, and activity of neutrophils and monocytes.","PeriodicalId":85139,"journal":{"name":"Medical immunology (London, England)","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67112336","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 : 2020-12-01DOI: 10.15789/1563-0625-nip-1978
V. V. Zhelezko, I. Novikova
The article presents the data on assessment of functional features of neutrophils in 34 patients with systemic lupus erythematosus (SLE). Development of neutrophil extracellular traps (NETs) was evaluated in cell cultures incubated in vitro for 30 and 150 minutes (basal levels, NETBAS30 and NETBAS150, respectively), and in the presence of heat-inactivated S. аureus (strain ATCC 25923, 108 CFU/ml) (stimulated levels, NETST30 and NETST150, respectively). NET looks like thin free-lying extracellular fibrillar structures, 2-3 times exceeding the size of unchanged granulocyte. The result was expressed as percentage and relative amount of extracellular traps per 100 counted leukocytes. Phagocytic activity of neutrophils was evaluated as phagocytosis of S. аureus by counting the percentage of neutrophils that engulfed phagocytic index of microbial particles (PI); the average number of phagocytosed objects per neutrophil phagocytic number (PC). ROS-producing activity was determined in the reduction of Nitroblue Tetrazolium tested in spontaneous and stimulated S. аureus variants (NBTBAS and NBTST, respectively). The result was expressed as the percentage of formazan-positive cells per 100 white blood cells. Nitroxide-producing properties were determined using the Crow (1999) method in spontaneous and stimulated samples for the accumulation of the nitrated amino acid tyrosine (3-nitrothyrosine, 3-NTBAS, and 3-NTST, respectively). We revealed a decrease in ROS production, phagocytosis and NO-forming activity of neutrophils associated with increased netosis. Activation of the netosis was observed in cell cultures without stimulation, indicating the in vivo formation of networks in SLE. The NET increase is most pronounced in the patients with lupus nephritis (p < 0.05), and in remission of the disease (p < 0.05). We have revealed a correlation of NET formation parameters with duration and degree of SLE activity (rs = -0.6; p = 0.001, and rs = 0.39; p = 0.02, respectively); autoantibody titers (anti-dsDNA and ANA) (rs = 0.67; р = 0.047 and rs = 0.59; р = 0.034, respectively); prothrombin complex activity (rs = 0.6; p = 0.036), as well and urea and creatinine levels (rs = 0.47; p = 0.037 and rs = 0.39; p = 0.048, respectively). The parameters of NETs can be considered a promising biomarker for verifying the diagnosis of SLE, evaluation of clinical activity, disease severity, and predicting the development of complications.
{"title":"Netosis in patients with systemic lupus erythematosus","authors":"V. V. Zhelezko, I. Novikova","doi":"10.15789/1563-0625-nip-1978","DOIUrl":"https://doi.org/10.15789/1563-0625-nip-1978","url":null,"abstract":"The article presents the data on assessment of functional features of neutrophils in 34 patients with systemic lupus erythematosus (SLE). Development of neutrophil extracellular traps (NETs) was evaluated in cell cultures incubated in vitro for 30 and 150 minutes (basal levels, NETBAS30 and NETBAS150, respectively), and in the presence of heat-inactivated S. аureus (strain ATCC 25923, 108 CFU/ml) (stimulated levels, NETST30 and NETST150, respectively). NET looks like thin free-lying extracellular fibrillar structures, 2-3 times exceeding the size of unchanged granulocyte. The result was expressed as percentage and relative amount of extracellular traps per 100 counted leukocytes. Phagocytic activity of neutrophils was evaluated as phagocytosis of S. аureus by counting the percentage of neutrophils that engulfed phagocytic index of microbial particles (PI); the average number of phagocytosed objects per neutrophil phagocytic number (PC). ROS-producing activity was determined in the reduction of Nitroblue Tetrazolium tested in spontaneous and stimulated S. аureus variants (NBTBAS and NBTST, respectively). The result was expressed as the percentage of formazan-positive cells per 100 white blood cells. Nitroxide-producing properties were determined using the Crow (1999) method in spontaneous and stimulated samples for the accumulation of the nitrated amino acid tyrosine (3-nitrothyrosine, 3-NTBAS, and 3-NTST, respectively). We revealed a decrease in ROS production, phagocytosis and NO-forming activity of neutrophils associated with increased netosis. Activation of the netosis was observed in cell cultures without stimulation, indicating the in vivo formation of networks in SLE. The NET increase is most pronounced in the patients with lupus nephritis (p < 0.05), and in remission of the disease (p < 0.05). We have revealed a correlation of NET formation parameters with duration and degree of SLE activity (rs = -0.6; p = 0.001, and rs = 0.39; p = 0.02, respectively); autoantibody titers (anti-dsDNA and ANA) (rs = 0.67; р = 0.047 and rs = 0.59; р = 0.034, respectively); prothrombin complex activity (rs = 0.6; p = 0.036), as well and urea and creatinine levels (rs = 0.47; p = 0.037 and rs = 0.39; p = 0.048, respectively). The parameters of NETs can be considered a promising biomarker for verifying the diagnosis of SLE, evaluation of clinical activity, disease severity, and predicting the development of complications.","PeriodicalId":85139,"journal":{"name":"Medical immunology (London, England)","volume":"22 1","pages":"987-992"},"PeriodicalIF":0.0,"publicationDate":"2020-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47678055","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 : 2020-12-01DOI: 10.15789/1563-0625-ita-2088
О. V. Berdyugina
Over recent years, the number of patients with tuberculosis has not decreased in the country and in worldwide. This is due to high resistance of the pathogen and changing mechanisms of bacterial perception by the human immune system thus requiring closer examination of the issue. Cell fusion during the formation of pulmonary tuberculous granuloma involves a large number of adhesive events. Importance of α1β1 integrin has been shown for the granuloma integrity during the chronic phase of infection. It has been proven that pulmonary tuberculous granuloma should be monitored, including with the detection of cells expressing CD11c, since they support the continuous priming of T cells at different stages of infection. The aim of this study was to answer the question, if there is a different expression of integrin receptors by immune cells from the patient’s peripheral blood at different stages of the existence of pulmonary tuberculous granuloma? The study involved 38 people: the first group (control) consisted of 15 practically healthy people; a second group included 11 subjects with pulmonary tuberculous granuloma; the condition was first diagnosed 2 to 10 months before the present study. A third group consisted of 12 patients with pulmonary tuberculous granuloma, with primary diagnosis established 12 to 219 months before this study. All the participants underwent a general clinical blood tests using a 5 Diff Mythic 22 AL analyzer (Cormay, Poland). The adhesion markers CD11b, CD11c were detected with a Coulter Epicx XL instrument (Beckman Coulter, USA). The following peripheral blood cell populations were determined: CD14- CD13lowCD11b+, CD14- CD13lowCD11c+, CD14+CD11b+, CD14+CD11c+, CD45+CD3- CD16+CD56+, CD45+CD3- CD16+CD56+CD11b+. Statistical processing of the results was performed in the Windows 10 operating environment (Microsoft Corp., USA), using Statistica v. 12.5 software (StatSoft, USA). Kruskal–Wallis one-way analysis of variance (pk-w), with differences significant at p < 0.017, as well as the Wald–Wolfowitz test (pw-w) at a significance level of p < 0.05 were used as criteria for assessing differences between the compared groups. In addition, cluster and factor analysis were implemented. When studying the role of β2-integrins, we have found that they play an important role in maintaining the existence of pulmonary tuberculous granuloma. An increase in total number of granulocytes, and CD11b-expressing granulocytes, a decrease in the population of lymphocytes, NK cells and NK cells expressing CD11c proved to be distinctive in cases of pulmonary tuberculous granuloma detected 0.5 years before the study. Characteristic changes observed in the study of peripheral blood in the patients with pulmonary tuberculous granuloma detected 9.5 years before the study were as follows: an increase in the leukocyte population, total monocyte number, as well as CD11band CD11c-expressing monocytes.
{"title":"Is there a dependence between expression of integrin receptors by peripheral blood immune cells and duration of tuberculous granuloma existence in the patients?","authors":"О. V. Berdyugina","doi":"10.15789/1563-0625-ita-2088","DOIUrl":"https://doi.org/10.15789/1563-0625-ita-2088","url":null,"abstract":"Over recent years, the number of patients with tuberculosis has not decreased in the country and in worldwide. This is due to high resistance of the pathogen and changing mechanisms of bacterial perception by the human immune system thus requiring closer examination of the issue. Cell fusion during the formation of pulmonary tuberculous granuloma involves a large number of adhesive events. Importance of α1β1 integrin has been shown for the granuloma integrity during the chronic phase of infection. It has been proven that pulmonary tuberculous granuloma should be monitored, including with the detection of cells expressing CD11c, since they support the continuous priming of T cells at different stages of infection. The aim of this study was to answer the question, if there is a different expression of integrin receptors by immune cells from the patient’s peripheral blood at different stages of the existence of pulmonary tuberculous granuloma? The study involved 38 people: the first group (control) consisted of 15 practically healthy people; a second group included 11 subjects with pulmonary tuberculous granuloma; the condition was first diagnosed 2 to 10 months before the present study. A third group consisted of 12 patients with pulmonary tuberculous granuloma, with primary diagnosis established 12 to 219 months before this study. All the participants underwent a general clinical blood tests using a 5 Diff Mythic 22 AL analyzer (Cormay, Poland). The adhesion markers CD11b, CD11c were detected with a Coulter Epicx XL instrument (Beckman Coulter, USA). The following peripheral blood cell populations were determined: CD14- CD13lowCD11b+, CD14- CD13lowCD11c+, CD14+CD11b+, CD14+CD11c+, CD45+CD3- CD16+CD56+, CD45+CD3- CD16+CD56+CD11b+. Statistical processing of the results was performed in the Windows 10 operating environment (Microsoft Corp., USA), using Statistica v. 12.5 software (StatSoft, USA). Kruskal–Wallis one-way analysis of variance (pk-w), with differences significant at p < 0.017, as well as the Wald–Wolfowitz test (pw-w) at a significance level of p < 0.05 were used as criteria for assessing differences between the compared groups. In addition, cluster and factor analysis were implemented. When studying the role of β2-integrins, we have found that they play an important role in maintaining the existence of pulmonary tuberculous granuloma. An increase in total number of granulocytes, and CD11b-expressing granulocytes, a decrease in the population of lymphocytes, NK cells and NK cells expressing CD11c proved to be distinctive in cases of pulmonary tuberculous granuloma detected 0.5 years before the study. Characteristic changes observed in the study of peripheral blood in the patients with pulmonary tuberculous granuloma detected 9.5 years before the study were as follows: an increase in the leukocyte population, total monocyte number, as well as CD11band CD11c-expressing monocytes.","PeriodicalId":85139,"journal":{"name":"Medical immunology (London, England)","volume":"22 1","pages":"867-878"},"PeriodicalIF":0.0,"publicationDate":"2020-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49345933","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 : 2020-12-01DOI: 10.15789/1563-0625-foc-2064
Lazareva Nm, O. Baranova, I. Kudryavtsev, N. A. Arsentieva, N. Liubimova, T. Ses’, M. Ilkovich, A. Totolian
Sarcoidosis is an inflammatory disease of unknown etiology with damage to the lungs and other organs characterized by development of necrosis-free epithelioid cell granulomas. Granulomatous inflammation characterized by the activation of different immune systems cells, in particular T lymphocytes, and the cytokines production. Our study was aimed at investigating the characteristics of the cytokine profile of blood plasma in patients with sarcoidosis. We studied peripheral blood plasma samples of patients with sarcoidosis (n = 52). The control blood samples were taken from healthy volunteers (n = 22). The level of 46 cytokines (pg/ml) was determined, as follows: IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL- 6, IL-7, IL-9, IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17A, IFNα2, IFNγ, TNFα, TNFβ, IL- 1ra, IL-10, EGF, FGF-2, Flt3 Ligand, G-CSF, GM-CSF, PDGF-AA, PDGF-AB / BB, TGFα, VEGF-A, sCD40L, CCL2, CCL3, CCL4, CCL5, CCL7, CCL11, CCL17, CCL20, CCL22, CXCL1, CXCL8, CXCL9, CXCL10, CXCL11, CXCL13, CX3CL1. Significantly higher levels of interleukins and some proinflammatory cytokines were found in the patients with sarcoidosis, i.e., IL-3, 0.70 vs 0.20, p = 0.003; IL-4, 14.37 vs 3.15, p = 0.009; IL-5, 1.06 vs 0.89, p < 0.001; IL-12 (p70), 1.27 vs 0.56, p = 0.028; IL-17A, 1.48 vs 0.43, p < 0.001; IFNα2, 41.79 vs 25.04, p = 0.003; IFNγ, 4.13 vs 1.14, p < 0.001; TNFα, 21.67 vs 6.70, p < 0.001; anti-inflammatory cytokine IL-10, 1.03 vs 0.45, p = 0.019; growth factors: FGF-2, 40.08 vs 30.58, p = 0.008, G-CSF, 24.18 vs 8.21, p = 0.006, and VEGF-A, 42.52 vs 26.76, p = 0.048; chemokines: CCL3, 3.86 vs 1.33, p < 0,001; CCL17, 78.24 vs 26.24, p < 0.001; CCL20, 7.19 vs 5.64, p = 0.021; CCL22, 660.60 vs 405.00, p < 0,001; CXCL9, 4013 vs 1142, p < 0,001; CXCL10, 565.90 vs 196.60, p < 0.001; CXCL11, 230.20 vs 121.10, p = 0.018; CX3CL1, 56.99 vs 5.16, p < 0.001. Peripheral blood chemokine CCL11 levels were significantly lower in patients compared to the group of healthy volunteers: 77.58 vs 124.70, p = 0.022. The features of the cytokine profile in patients with sarcoidosis may indicate their important role in the processes of formation and outcomes of granulomas. These issues require an additional detailed study, comparison with phenotypes, differential course and outcomes of the disease.
结节病是一种病因不明的炎症性疾病,对肺部和其他器官造成损害,其特征是形成无坏死的上皮样细胞肉芽肿。以激活不同免疫系统细胞,特别是T淋巴细胞和产生细胞因子为特征的肉芽肿性炎症。我们的研究旨在研究结节病患者血浆中细胞因子谱的特征。我们研究了结节病患者(n=52)的外周血血浆样本。对照血样取自健康志愿者(n=22)。测定46种细胞因子的水平(pg/ml),如下:IL-1α、IL-1β、IL-2、IL-3、IL-4、IL-5、IL-6、IL-7、IL-9、IL-12(p40)、IL-12、p70、IL-13、IL-15、IL-17A、IFNα2、IFNγ、TNFα、TNFβ、IL-11a、IL-10、EGF、FGF-2、Flt3配体、G-CSF、GM-CSF、PDGF-AA、PDGF-AB/BB、TGFα、VEGF-A、sCD40L、CCL2、CCL3、CCL4、CCL5、CCL7。CCL11、CCL17、CCL20、CCL22、CXCL1、CXCL8、CXCL9、CXCL10、CXCL11,CXCL13、CX3CL1。结节病患者的白细胞介素和一些促炎细胞因子水平显著较高,即IL-3,0.70比0.20,p=0.003;IL-4,14.37 vs 3.15,p=0.009;IL-5,1.06 vs 0.89,p<0.001;IL-12(p70),1.27 vs 0.56,p=0.028;IL-17A,1.48 vs 0.43,p<0.001;IFNα2,41.79 vs 25.04,p=0.003;IFNγ,4.13 vs 1.14,p<0.001;TNFα,21.67 vs 6.70,p<0.001;抗炎细胞因子IL-10,1.03 vs 0.45,p=0.019;生长因子:FGF-2,40.08 vs 30.58,p=0.008;G-CSF,24.18 vs 8.21,p=0.006;VEGF-A,42.52 vs 26.76,p=0.048;趋化因子:CCL3,3.86 vs 1.33,p<0001;CCL17,78.24 vs 26.24,p<0.001;CCL20,7.19对5.64,p=0.021;CCL22660.60对405.00,p<0001;CXCL9,4013对1142,p<0001;CXCL10,565.90 vs 196.60,p<0.001;CXCL11230.20对121.10,p=0.018;CX3CL1,56.99 vs 5.16,p<0.001。与健康志愿者组相比,患者的外周血趋化因子CCL11水平显著降低:77.58比124.70,p=0.022。结节病患者细胞因子谱的特征可能表明它们在肉芽肿的形成过程和结果中的重要作用。这些问题需要额外的详细研究,与表型、不同病程和疾病结果进行比较。
{"title":"Features of cytokine profile in patients with sarcoidosis","authors":"Lazareva Nm, O. Baranova, I. Kudryavtsev, N. A. Arsentieva, N. Liubimova, T. Ses’, M. Ilkovich, A. Totolian","doi":"10.15789/1563-0625-foc-2064","DOIUrl":"https://doi.org/10.15789/1563-0625-foc-2064","url":null,"abstract":"Sarcoidosis is an inflammatory disease of unknown etiology with damage to the lungs and other organs characterized by development of necrosis-free epithelioid cell granulomas. Granulomatous inflammation characterized by the activation of different immune systems cells, in particular T lymphocytes, and the cytokines production. Our study was aimed at investigating the characteristics of the cytokine profile of blood plasma in patients with sarcoidosis. We studied peripheral blood plasma samples of patients with sarcoidosis (n = 52). The control blood samples were taken from healthy volunteers (n = 22). The level of 46 cytokines (pg/ml) was determined, as follows: IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL- 6, IL-7, IL-9, IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17A, IFNα2, IFNγ, TNFα, TNFβ, IL- 1ra, IL-10, EGF, FGF-2, Flt3 Ligand, G-CSF, GM-CSF, PDGF-AA, PDGF-AB / BB, TGFα, VEGF-A, sCD40L, CCL2, CCL3, CCL4, CCL5, CCL7, CCL11, CCL17, CCL20, CCL22, CXCL1, CXCL8, CXCL9, CXCL10, CXCL11, CXCL13, CX3CL1. Significantly higher levels of interleukins and some proinflammatory cytokines were found in the patients with sarcoidosis, i.e., IL-3, 0.70 vs 0.20, p = 0.003; IL-4, 14.37 vs 3.15, p = 0.009; IL-5, 1.06 vs 0.89, p < 0.001; IL-12 (p70), 1.27 vs 0.56, p = 0.028; IL-17A, 1.48 vs 0.43, p < 0.001; IFNα2, 41.79 vs 25.04, p = 0.003; IFNγ, 4.13 vs 1.14, p < 0.001; TNFα, 21.67 vs 6.70, p < 0.001; anti-inflammatory cytokine IL-10, 1.03 vs 0.45, p = 0.019; growth factors: FGF-2, 40.08 vs 30.58, p = 0.008, G-CSF, 24.18 vs 8.21, p = 0.006, and VEGF-A, 42.52 vs 26.76, p = 0.048; chemokines: CCL3, 3.86 vs 1.33, p < 0,001; CCL17, 78.24 vs 26.24, p < 0.001; CCL20, 7.19 vs 5.64, p = 0.021; CCL22, 660.60 vs 405.00, p < 0,001; CXCL9, 4013 vs 1142, p < 0,001; CXCL10, 565.90 vs 196.60, p < 0.001; CXCL11, 230.20 vs 121.10, p = 0.018; CX3CL1, 56.99 vs 5.16, p < 0.001. Peripheral blood chemokine CCL11 levels were significantly lower in patients compared to the group of healthy volunteers: 77.58 vs 124.70, p = 0.022. The features of the cytokine profile in patients with sarcoidosis may indicate their important role in the processes of formation and outcomes of granulomas. These issues require an additional detailed study, comparison with phenotypes, differential course and outcomes of the disease.","PeriodicalId":85139,"journal":{"name":"Medical immunology (London, England)","volume":"22 1","pages":"993-1002"},"PeriodicalIF":0.0,"publicationDate":"2020-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45612652","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 : 2020-12-01DOI: 10.15789/1563-0625-roi-1986
S. Tereschenko, M. Smolnikova, E. Kasparov, E. Shakhtshneider, M. A. Malinchik, O. Konopleva, S. Smirnova
Bronchial asthma is a multifactorial disease, with both environmental factors and genetic predisposal affecting its development. A number of gene associations have been obtained between polymorphisms of cytokine genes produced by different types of immune cells and asthma development. Interleukin-13 is involved in allergic inflammation, increased bronchial hypersensitivity, regulation of eosinophil levels and IgE production by B cells, thus making it promising for studying IL13 gene polymorphisms in bronchial asthma coupled to development of the disease. The aim of this study was to investigate possible association between asthma and IL13 rs1800925 polymorphism in the children of Caucasian origin in Eastern Siberia. Four groups of patients with asthma were examined (mean age 12.8±1.2 years): with a controlled (n = 95) and uncontrolled course (n = 107), with severe (n = 71) and moderate severity (n = 131) diseases. The control group consisted of healthy individuals: children (n = 33) and adults (n = 102). DNA was isolated with sorbent method; genotyping was carried out using RT-PCR using specific oligonucleotide primers and fluorescent TaqMan probes. The allele and genotype frequencies were compared by the χ-square test using an online calculator. The odds ratio (OR) with a 95% confidence interval (CI) was performed to link genetic markers with pathological phenotypes. The CT IL13 rs1800925 genotype was shown to be associated with moderate asthma and cases of uncontrollable clinical course, whereas the TT genotype was associated with severe asthma. Thus, rs1800925 polymorphism of IL13 gene (the T* variant is known to be associated with increased IL-13 expression) may be associated with bronchial asthma in children. Our data are consistent with results of other authors. E.g., Liu Z. et al. revealed an association between rs1800925 IL13 and the risk of developing asthma in children, with CT and TT genotypes being more common in the patient group. Radhakrishnan A. et al., was studied rs1800925 IL13 in adult population of Malaysia and found that the T* allele frequency in the group of patients significantly exceeds the frequency of this allele in the control group. Thus, the results of our study showed that IL13 rs1800925 polymorphism is associated with bronchial asthma in children, especially, with level of its control and severity of the disease.
{"title":"Role of IL13 genetic polymorphism in the development of bronchial asthma in children","authors":"S. Tereschenko, M. Smolnikova, E. Kasparov, E. Shakhtshneider, M. A. Malinchik, O. Konopleva, S. Smirnova","doi":"10.15789/1563-0625-roi-1986","DOIUrl":"https://doi.org/10.15789/1563-0625-roi-1986","url":null,"abstract":"Bronchial asthma is a multifactorial disease, with both environmental factors and genetic predisposal affecting its development. A number of gene associations have been obtained between polymorphisms of cytokine genes produced by different types of immune cells and asthma development. Interleukin-13 is involved in allergic inflammation, increased bronchial hypersensitivity, regulation of eosinophil levels and IgE production by B cells, thus making it promising for studying IL13 gene polymorphisms in bronchial asthma coupled to development of the disease. The aim of this study was to investigate possible association between asthma and IL13 rs1800925 polymorphism in the children of Caucasian origin in Eastern Siberia. Four groups of patients with asthma were examined (mean age 12.8±1.2 years): with a controlled (n = 95) and uncontrolled course (n = 107), with severe (n = 71) and moderate severity (n = 131) diseases. The control group consisted of healthy individuals: children (n = 33) and adults (n = 102). DNA was isolated with sorbent method; genotyping was carried out using RT-PCR using specific oligonucleotide primers and fluorescent TaqMan probes. The allele and genotype frequencies were compared by the χ-square test using an online calculator. The odds ratio (OR) with a 95% confidence interval (CI) was performed to link genetic markers with pathological phenotypes. The CT IL13 rs1800925 genotype was shown to be associated with moderate asthma and cases of uncontrollable clinical course, whereas the TT genotype was associated with severe asthma. Thus, rs1800925 polymorphism of IL13 gene (the T* variant is known to be associated with increased IL-13 expression) may be associated with bronchial asthma in children. Our data are consistent with results of other authors. E.g., Liu Z. et al. revealed an association between rs1800925 IL13 and the risk of developing asthma in children, with CT and TT genotypes being more common in the patient group. Radhakrishnan A. et al., was studied rs1800925 IL13 in adult population of Malaysia and found that the T* allele frequency in the group of patients significantly exceeds the frequency of this allele in the control group. Thus, the results of our study showed that IL13 rs1800925 polymorphism is associated with bronchial asthma in children, especially, with level of its control and severity of the disease.","PeriodicalId":85139,"journal":{"name":"Medical immunology (London, England)","volume":"22 1","pages":"907-914"},"PeriodicalIF":0.0,"publicationDate":"2020-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44712394","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 : 2020-12-01DOI: 10.15789/1563-0625-sfg-2050
M. Smolnikova, A. A. Barilo, M. A. Malinchik, S. Smirnova
Psoriasis (PS) and psoriatic arthritis (PsA) are interrelated diseases that occur in approximately 30% of patients and are characterized by the presence of a systemic inflammatory reaction that occurs as a result of a violation of the functional state of the immune system. With the advent of new technologies, several new pro-inflammatory cytokines, such as IL-23, IL-31, and IL-33, which play an important role in the pathogenesis of the psoriatic process, have been discovered and characterized. It was determined that single nucleotide polymorphisms (SNPs) in the promoter regions of the IL23, IL31 and IL33 genes play an important role in controlling the expression of relevant cytokines involved in the immunopathogenesis of psoriatic disease. The purpose of the study: to analyze the distribution of genotypes and allelic variants of polymorphisms of the IL23A (rs2066808), IL23R (rs2201841), IL31 (rs7977932) and IL33 (rs7044343), in order to search for genetic markers of predisposition to psoriasis and psoriatic arthritis. Materials and methods. The genotyping of the patients was conducted: psoriasis (PS, n = 77), median age 31.0 years (27.0-43.0), psoriatic arthritis (PsA, n = 99), median age 49.0 years (39.0-56.0) and practically healthy residents of Krasnoyarsk (n = 103), a median age of 32.0 years (24.0-38.0). DNA was isolated from whole venous blood using a standard sorbent kit. Genotyping of single nucleotide polymorphisms IL23A (rs2066808), IL23R (rs2201841), IL31 (rs7977932), IL33 (rs7044343) was carried out using real-time PCR using specific oligonucleotide primers and fluorescentlylabeled probes. Results and discussion. The frequencies of allelic variants of the studied cytokine genes in the control group obtained during the study correspond to their distribution in Caucasoid populations – the alleles IL23A * T, IL23R * T, IL31 * C, IL33 * C prevail. When comparing the distribution frequency of allelic variants of the IL23A, IL23R, IL31, IL33 genes, we did not obtain statistically significant differences between patients and the control group. Conclusions. Despite the fact that when comparing the distribution frequency of allelic variants of the IL23A, IL23R, IL31, IL33 genes, we did not obtain statistically significant differences between the patients and the control group, there are results worthy of attention. So, in patients with PS, the frequency of the C * IL23A allelic variant (rs2066808) is lower than in the population sample, which may indicate its specific role in relation to the development of the disease. All this dictates the need to continue research with the assessment of other SNPs and increase the sample of patients in search of potential genetic markers of psoriatic disease.
{"title":"Search for genetic markers of predisposition to psoriasis and psoriatic arthritis","authors":"M. Smolnikova, A. A. Barilo, M. A. Malinchik, S. Smirnova","doi":"10.15789/1563-0625-sfg-2050","DOIUrl":"https://doi.org/10.15789/1563-0625-sfg-2050","url":null,"abstract":"Psoriasis (PS) and psoriatic arthritis (PsA) are interrelated diseases that occur in approximately 30% of patients and are characterized by the presence of a systemic inflammatory reaction that occurs as a result of a violation of the functional state of the immune system. With the advent of new technologies, several new pro-inflammatory cytokines, such as IL-23, IL-31, and IL-33, which play an important role in the pathogenesis of the psoriatic process, have been discovered and characterized. It was determined that single nucleotide polymorphisms (SNPs) in the promoter regions of the IL23, IL31 and IL33 genes play an important role in controlling the expression of relevant cytokines involved in the immunopathogenesis of psoriatic disease. The purpose of the study: to analyze the distribution of genotypes and allelic variants of polymorphisms of the IL23A (rs2066808), IL23R (rs2201841), IL31 (rs7977932) and IL33 (rs7044343), in order to search for genetic markers of predisposition to psoriasis and psoriatic arthritis. Materials and methods. The genotyping of the patients was conducted: psoriasis (PS, n = 77), median age 31.0 years (27.0-43.0), psoriatic arthritis (PsA, n = 99), median age 49.0 years (39.0-56.0) and practically healthy residents of Krasnoyarsk (n = 103), a median age of 32.0 years (24.0-38.0). DNA was isolated from whole venous blood using a standard sorbent kit. Genotyping of single nucleotide polymorphisms IL23A (rs2066808), IL23R (rs2201841), IL31 (rs7977932), IL33 (rs7044343) was carried out using real-time PCR using specific oligonucleotide primers and fluorescentlylabeled probes. Results and discussion. The frequencies of allelic variants of the studied cytokine genes in the control group obtained during the study correspond to their distribution in Caucasoid populations – the alleles IL23A * T, IL23R * T, IL31 * C, IL33 * C prevail. When comparing the distribution frequency of allelic variants of the IL23A, IL23R, IL31, IL33 genes, we did not obtain statistically significant differences between patients and the control group. Conclusions. Despite the fact that when comparing the distribution frequency of allelic variants of the IL23A, IL23R, IL31, IL33 genes, we did not obtain statistically significant differences between the patients and the control group, there are results worthy of attention. So, in patients with PS, the frequency of the C * IL23A allelic variant (rs2066808) is lower than in the population sample, which may indicate its specific role in relation to the development of the disease. All this dictates the need to continue research with the assessment of other SNPs and increase the sample of patients in search of potential genetic markers of psoriatic disease.","PeriodicalId":85139,"journal":{"name":"Medical immunology (London, England)","volume":"22 1","pages":"925-932"},"PeriodicalIF":0.0,"publicationDate":"2020-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48516955","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 : 2020-12-01DOI: 10.15789/1563-0625-eoi-2034
A. Miromanov, T. Zabello, N. Miromanova
Our objective was to study the effects of IL4-589C>T, FCGR2A-166His>Arg, DEFB1-20G>A, DEFB1-52G>A gene polymorphisms upon content of TNFα, IL-1β, IL-4, and IL-10 in primary osteoarthrosis of the hip joints. We performed a survey of 100 patients of Russian ethnicity (average age 61.3±8.5 years) with primary coxarthrosis at the stage III-IV who lived in the Trans-Baikal region. The control group (n = 100), were local residents, comparable by age (60±8.3 years), gender, habitation place and nationality. The exclusion criteria were as follows: close relationship; other types of osteoarthritis (post-traumatic, rheumatoid, metabolic, etc.); dysplastic syndromes and phenotypes; acute and chronic inflammatory diseases at the exacerbation stage; diabetes mellitus; osteoporosis; vascular diseases; obesity; malignant neoplasia; alcohol abuse. Along with clinical examination, the following laboratory methods were applied: immunological techniques, i.e., determination of TNFα, IL-1β, IL-4, IL-10; genetic testing using polymerase chain reaction, e.g., a point mutation of the IL4 gene at the 589(C>T) position, FCGR2A at 166(His>Arg) site, DEFB1 at the 20(G>A) and 52(G>A) positions. DNA from the peripheral blood of patients was used for the molecular genetic analysis. Radiographic examination was also carried out. The data were statistically processed using STATISTICA 6.1 software package (StatSoft, USA), Microsoft Office Excel 2019 for Windows 10. The differences were considered statistically significant at p ≤ 0.05. Results. The -589T/T genotype of IL4-589C>T gene polymorphism indirectly contributes to higher content of TNFα and IL-1β for primary osteoarthritis of the hip joints. The patients with -166Arg/Arg genotype have a 1.3-fold increase of certain cytokine concentrations, e.g., TNFα and IL-1β, as compared with -166His/Arg genotype, and, conversely, lower content of IL-4 and IL-10 (1.3- fold) in comparison with -166His/His genotype. The patients with -20A/A genotype showed higher levels of TNFα and IL-1β, respectively, 1.2 and 1.3 times, compared with -20G/G genotype, and 1.3 times versus the -20G/A genotype. Conclusions: 1. The presence of -589T/T genotype of the IL4-589C>T gene polymorphism and the -20A/A genotype of the DEFB1-20G>A gene polymorphism contributes to a high content of TNFα and IL-1β in the blood serum, and the carriage of -166His/His FCGR2A-166His>Arg gene polymorphism is associated with both higher level of TNFα, IL-1β, and a low concentration of IL-4, IL-10. 2. Complex carriers of FCGR2A166HisArg x DEFB152AA x DEFB120AA x IL4589TT genotypes in the patients with primary coxarthrosis increases the contents of TNFα, IL-1β cytokines by 1.5 and 1.7 times, respectively.
我们的目的是研究IL4-589C >t、FCGR2A-166His >arg、DEFB1-20G >a、DEFB1-52G >a基因多态性对原发性髋关节骨关节病患者TNFα、IL-1β、IL-4和IL-10含量的影响。我们对100名居住在贝加尔地区的俄罗斯族(平均年龄61.3±8.5岁)III-IV期原发性关节关节病患者进行了调查。对照组(n = 100)为当地居民,年龄(60±8.3岁)、性别、居住地、民族具有可比性。排除标准为:关系密切;其他类型的骨关节炎(创伤后、类风湿、代谢性等);发育不良综合征和表型;急性加重期急慢性炎性疾病;糖尿病;骨质疏松症;血管疾病;肥胖;恶性肿瘤;酒精滥用。结合临床检查,采用以下实验室方法:免疫学技术,即检测TNFα、IL-1β、IL-4、IL-10;利用聚合酶链反应进行基因检测,例如,il - 4基因在589(C>T)位点、FCGR2A在166(His>Arg)位点、DEFB1在20(G> a)和52(G> a)位点发生点突变。从患者外周血中提取DNA用于分子遗传学分析。同时进行x线检查。使用STATISTICA 6.1软件包(StatSoft, USA)、Microsoft Office Excel 2019 for Windows 10对数据进行统计处理。p≤0.05认为差异有统计学意义。结果。IL4-589C >t基因多态性的-589T/T基因型间接导致原发性髋关节骨关节炎中TNFα和IL-1β含量升高。与- 166his /Arg基因型患者相比,- 166arg /Arg基因型患者某些细胞因子如TNFα和IL-1β浓度增加1.3倍,相反,IL-4和IL-10含量较- 166his /His基因型低(1.3倍)。-20A/A基因型患者的TNFα和IL-1β水平分别比-20G/G基因型高1.2倍和1.3倍,比-20G/A基因型高1.3倍。结论:1。-589T/T基因型的IL4-589C>T基因多态性和-20A/A基因型的DEFB1-20G>A基因多态性的存在导致血清中TNFα和IL-1β含量高,-166His/His FCGR2A-166His>Arg基因多态性的携带与TNFα、IL-1β水平高和IL-4、IL-10浓度低有关。2. 原发性关节关节病患者中携带FCGR2A166HisArg x DEFB152AA x DEFB120AA x IL4589TT基因型的复合携带者,其TNFα、IL-1β细胞因子含量分别增加1.5倍和1.7倍。
{"title":"Effect of IL4-589C>T, FCGR2A-166His>Arg, DEFB1-20G>A, DEFB1-52G>A gene polymorphisms on TNFα, IL-1β, IL-4, and IL-10 contents in the patients with primary hip osteoarthrosis","authors":"A. Miromanov, T. Zabello, N. Miromanova","doi":"10.15789/1563-0625-eoi-2034","DOIUrl":"https://doi.org/10.15789/1563-0625-eoi-2034","url":null,"abstract":"Our objective was to study the effects of IL4-589C>T, FCGR2A-166His>Arg, DEFB1-20G>A, DEFB1-52G>A gene polymorphisms upon content of TNFα, IL-1β, IL-4, and IL-10 in primary osteoarthrosis of the hip joints. We performed a survey of 100 patients of Russian ethnicity (average age 61.3±8.5 years) with primary coxarthrosis at the stage III-IV who lived in the Trans-Baikal region. The control group (n = 100), were local residents, comparable by age (60±8.3 years), gender, habitation place and nationality. The exclusion criteria were as follows: close relationship; other types of osteoarthritis (post-traumatic, rheumatoid, metabolic, etc.); dysplastic syndromes and phenotypes; acute and chronic inflammatory diseases at the exacerbation stage; diabetes mellitus; osteoporosis; vascular diseases; obesity; malignant neoplasia; alcohol abuse. Along with clinical examination, the following laboratory methods were applied: immunological techniques, i.e., determination of TNFα, IL-1β, IL-4, IL-10; genetic testing using polymerase chain reaction, e.g., a point mutation of the IL4 gene at the 589(C>T) position, FCGR2A at 166(His>Arg) site, DEFB1 at the 20(G>A) and 52(G>A) positions. DNA from the peripheral blood of patients was used for the molecular genetic analysis. Radiographic examination was also carried out. The data were statistically processed using STATISTICA 6.1 software package (StatSoft, USA), Microsoft Office Excel 2019 for Windows 10. The differences were considered statistically significant at p ≤ 0.05. Results. The -589T/T genotype of IL4-589C>T gene polymorphism indirectly contributes to higher content of TNFα and IL-1β for primary osteoarthritis of the hip joints. The patients with -166Arg/Arg genotype have a 1.3-fold increase of certain cytokine concentrations, e.g., TNFα and IL-1β, as compared with -166His/Arg genotype, and, conversely, lower content of IL-4 and IL-10 (1.3- fold) in comparison with -166His/His genotype. The patients with -20A/A genotype showed higher levels of TNFα and IL-1β, respectively, 1.2 and 1.3 times, compared with -20G/G genotype, and 1.3 times versus the -20G/A genotype. Conclusions: 1. The presence of -589T/T genotype of the IL4-589C>T gene polymorphism and the -20A/A genotype of the DEFB1-20G>A gene polymorphism contributes to a high content of TNFα and IL-1β in the blood serum, and the carriage of -166His/His FCGR2A-166His>Arg gene polymorphism is associated with both higher level of TNFα, IL-1β, and a low concentration of IL-4, IL-10. 2. Complex carriers of FCGR2A166HisArg x DEFB152AA x DEFB120AA x IL4589TT genotypes in the patients with primary coxarthrosis increases the contents of TNFα, IL-1β cytokines by 1.5 and 1.7 times, respectively.","PeriodicalId":85139,"journal":{"name":"Medical immunology (London, England)","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67109367","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 : 2020-12-01DOI: 10.15789/1563-0625-uct-2058
D. G. Ponomarenko, E. Rakitina, M. Kostyuchenko, O. Logvinenko, A. Ryazanova, L. Aksenova, N. P. Buravtseva, I. Tyumentseva, S. A. Kurcheva, A. Kulichenko
We present the results of applying functional cytometric test of antigen-stimulated activation basophils to assess specific immunological reactivity in the people with anthrax, and immunized with anthrax vaccine. As a criterion for antigen-specific basophil activation, we measured expression of the CD63 membrane receptor, which reflects the process of anaphylactic basophil degranulation. To determine spontaneous and antigen-induced activation of basophils (CCR3+CD63+), a FlowCAST reagent kit (Buhlmann laboratories AG, Switzerland) was used. Anthraxin, an experimental anthrax allergen (a hydrolysate the Bacillus anthracis STI-1 strain), manufactured by the Stavropol Anti-Plague Institute, was used as a specific antigen. As based on clinical and experimental data, a threshold value of > 10% of anthraxin-activated (CCR3+CD63+) basophils was accepted for the in vitro immunodiagnostic CAST test, as a laboratory criterion for the subjects exhibiting specific immune response, i.e., IgE-mediated sensitization. It was shown that, in anthrax patients within one week after onset of the disease (3-7 days), a positive CAST result was obtained in 92.3% cases; the levels of specific basophil activation with anthraxin averaged 37.9% (12.01 ÷ 78.9%). Immunological examination of individuals three weeks (21 days) after vaccination against anthrax revealed CAST-positivity in all the vaccinated persons. Intensity of anthraxin-induced basophil activation the vaccinated subjects was ranged from 10.87 to 30.03%, averaging 17.86%. The overall values of spontaneous and specific activation ranged within 12.39 ÷ 41.46%. The study opens prospectives for implementation of basophil antigenic activation test in the Flow CAST format in diagnostics of anthrax and to identify specific immune rearrangements after vaccination in humans, as an index of actual vaccination rates. Usage of CAST test with anthraxin makes it possible to identify anthrax patients at the early stages (2-4 days after onset of the disease) including, among patients with an increased CCR3+CD63+ background values, evaluation of immunological efficiency in the cohorts at risk for vaccination. At the same time, it was found that a significant decrease in diagnostic sensitivity of CAST test could be observed in the patients immune to anthrax pathogen who received intensive antibacterial and pathogenetic therapy at the early stages of infection, including glucocorticosteroids (anti-inflammatory drugs) and desensitizing agents that inhibit the degree of hypersensitivity development and its expression.
{"title":"Using CAST-test to investigate human specific hypersensitivity to the anthrax pathogen","authors":"D. G. Ponomarenko, E. Rakitina, M. Kostyuchenko, O. Logvinenko, A. Ryazanova, L. Aksenova, N. P. Buravtseva, I. Tyumentseva, S. A. Kurcheva, A. Kulichenko","doi":"10.15789/1563-0625-uct-2058","DOIUrl":"https://doi.org/10.15789/1563-0625-uct-2058","url":null,"abstract":"We present the results of applying functional cytometric test of antigen-stimulated activation basophils to assess specific immunological reactivity in the people with anthrax, and immunized with anthrax vaccine. As a criterion for antigen-specific basophil activation, we measured expression of the CD63 membrane receptor, which reflects the process of anaphylactic basophil degranulation. To determine spontaneous and antigen-induced activation of basophils (CCR3+CD63+), a FlowCAST reagent kit (Buhlmann laboratories AG, Switzerland) was used. Anthraxin, an experimental anthrax allergen (a hydrolysate the Bacillus anthracis STI-1 strain), manufactured by the Stavropol Anti-Plague Institute, was used as a specific antigen. As based on clinical and experimental data, a threshold value of > 10% of anthraxin-activated (CCR3+CD63+) basophils was accepted for the in vitro immunodiagnostic CAST test, as a laboratory criterion for the subjects exhibiting specific immune response, i.e., IgE-mediated sensitization. It was shown that, in anthrax patients within one week after onset of the disease (3-7 days), a positive CAST result was obtained in 92.3% cases; the levels of specific basophil activation with anthraxin averaged 37.9% (12.01 ÷ 78.9%). Immunological examination of individuals three weeks (21 days) after vaccination against anthrax revealed CAST-positivity in all the vaccinated persons. Intensity of anthraxin-induced basophil activation the vaccinated subjects was ranged from 10.87 to 30.03%, averaging 17.86%. The overall values of spontaneous and specific activation ranged within 12.39 ÷ 41.46%. The study opens prospectives for implementation of basophil antigenic activation test in the Flow CAST format in diagnostics of anthrax and to identify specific immune rearrangements after vaccination in humans, as an index of actual vaccination rates. Usage of CAST test with anthraxin makes it possible to identify anthrax patients at the early stages (2-4 days after onset of the disease) including, among patients with an increased CCR3+CD63+ background values, evaluation of immunological efficiency in the cohorts at risk for vaccination. At the same time, it was found that a significant decrease in diagnostic sensitivity of CAST test could be observed in the patients immune to anthrax pathogen who received intensive antibacterial and pathogenetic therapy at the early stages of infection, including glucocorticosteroids (anti-inflammatory drugs) and desensitizing agents that inhibit the degree of hypersensitivity development and its expression.","PeriodicalId":85139,"journal":{"name":"Medical immunology (London, England)","volume":"22 1","pages":"1017-1024"},"PeriodicalIF":0.0,"publicationDate":"2020-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47907089","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 : 2020-12-01DOI: 10.15789/1563-0625-pal-2002
V. Neroev, N. Balatskaya, A. Novikova, M. V. Ryabina, P. A. Ilyukhin
Pathogenesis of retinal capillary hemangioma has not been sufficiently studied at the present time. Therefore, the study of cytokine levels in biological fluids seems to be very relevant in order to increase knowledge about the mechanisms of the disease development and searching for targeted therapies. The content of hematopoietic and vasoactive growth factors in blood serum, lacrimal fluid, and vitreous body was studied in patients with retinal capillary hemangioma. A total of 26 patients with retinal angiomatosis were examined. The samples of blood serum (n = 23) and lacrimal fluid (n = 10) from practically healthy people aged 22 to 46 (27.4±1.4 years) were used as a control. To perform comparative assessment of cytokine concentrations in the vitreous body of patients with retinal capillary hemangioma, were used samples of the vitreous body from 6 patients (average age 33±4.7 years; from 21 to 49 years) with rhegmatogenous retinal detachment. To measure the cytokine concentrations, we applied multiplex analysis technique using the xMAP platform with LuminexxPONENT 3.1 program and ProcartaPlex sets (eBioscience, Austria). A detailed characteristic of vasoactive factors in capillary retinal hemangioma was obtained as a result of this work. Some disorders in chemokine regulation were identified. There was a significant increase in serum concentrations of three vasoactive factors, i.e., PDGF-BB, HGF, and PIGF-1, with a decrease in chemokines (MCP-1, MIP-1α, and MIP-1β). The frequencies of PIGF-1 and MIP-1α detection also significantly differed from the control group. SCF was significantly more often determined in patients with retinal angiomatosis only at the systemic level. Correlations between PDGF-BB and PIGF-1, as well as PIGF-1 and MIP-1β were shown. A significant increase in VEGF-A, HGF, VEGF-D, as well as MCP-1 concentrations was shown in the lacrimal fluid. The inversion of PDGF-BB concentrations in serum and lacrimal fluid was noted. Analysis of intraocular cytokine levels revealed a significant increase in VEGF-A and HGF concentrations, with marked decrease in MIP-1α and MIP-1β. PDGF-BB in 100% of cases was determined only in vitreous body of patients with retinal angiomatosis. With respect to the revealed characteristic shifts of HGF/SF intraocular production in retinal capillary hemangioma, it seems relevant to search ways for its inhibition, thus providing potential basis for a new therapeutic strategy in treatment of retinal angiomatosis.
{"title":"Proven and less studied hematopoietic and vasoactive growth factors in retinal capillary hemangioma","authors":"V. Neroev, N. Balatskaya, A. Novikova, M. V. Ryabina, P. A. Ilyukhin","doi":"10.15789/1563-0625-pal-2002","DOIUrl":"https://doi.org/10.15789/1563-0625-pal-2002","url":null,"abstract":"Pathogenesis of retinal capillary hemangioma has not been sufficiently studied at the present time. Therefore, the study of cytokine levels in biological fluids seems to be very relevant in order to increase knowledge about the mechanisms of the disease development and searching for targeted therapies. The content of hematopoietic and vasoactive growth factors in blood serum, lacrimal fluid, and vitreous body was studied in patients with retinal capillary hemangioma. A total of 26 patients with retinal angiomatosis were examined. The samples of blood serum (n = 23) and lacrimal fluid (n = 10) from practically healthy people aged 22 to 46 (27.4±1.4 years) were used as a control. To perform comparative assessment of cytokine concentrations in the vitreous body of patients with retinal capillary hemangioma, were used samples of the vitreous body from 6 patients (average age 33±4.7 years; from 21 to 49 years) with rhegmatogenous retinal detachment. To measure the cytokine concentrations, we applied multiplex analysis technique using the xMAP platform with LuminexxPONENT 3.1 program and ProcartaPlex sets (eBioscience, Austria). A detailed characteristic of vasoactive factors in capillary retinal hemangioma was obtained as a result of this work. Some disorders in chemokine regulation were identified. There was a significant increase in serum concentrations of three vasoactive factors, i.e., PDGF-BB, HGF, and PIGF-1, with a decrease in chemokines (MCP-1, MIP-1α, and MIP-1β). The frequencies of PIGF-1 and MIP-1α detection also significantly differed from the control group. SCF was significantly more often determined in patients with retinal angiomatosis only at the systemic level. Correlations between PDGF-BB and PIGF-1, as well as PIGF-1 and MIP-1β were shown. A significant increase in VEGF-A, HGF, VEGF-D, as well as MCP-1 concentrations was shown in the lacrimal fluid. The inversion of PDGF-BB concentrations in serum and lacrimal fluid was noted. Analysis of intraocular cytokine levels revealed a significant increase in VEGF-A and HGF concentrations, with marked decrease in MIP-1α and MIP-1β. PDGF-BB in 100% of cases was determined only in vitreous body of patients with retinal angiomatosis. With respect to the revealed characteristic shifts of HGF/SF intraocular production in retinal capillary hemangioma, it seems relevant to search ways for its inhibition, thus providing potential basis for a new therapeutic strategy in treatment of retinal angiomatosis.","PeriodicalId":85139,"journal":{"name":"Medical immunology (London, England)","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67110544","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 : 2020-11-27DOI: 10.15789/1563-0625-iam-2037
A. Savchenko, A. Borisov, I. Kudryavtsev, I. Gvozdev, A. Moshev
The aim of the present study was to analyze the relationships between expression of activation and adhesion receptors on peripheral blood neutrophils, and intracellular activity of some neutrophil enzymes in patients with kidney cancer (KC). Patients and methods: the KC patients (n = 72) (T3N0M0, clear-cell type) were examined prior to surgical treatment at the Krasnoyarsk Regional Oncology Center. The diagnosis was verified histologically for all KC patients. The phenotype of blood neutrophils was studied using flow cytometry. The surface receptor expression levels of the neutrophils were evaluated by mean fluorescence intensity. NAD and NADP-dependent dehydrogenases activities in purified peripheral blood neutrophils were measured by bioluminescent method. Results: we have found that the phenotypic alterations in circulating KC patients’ neutrophils appeared along with inhibition of main intracellular metabolic processes and were closely linked with them. The features of the phenotypic imbalance in the neutrophils from KC patients were associated with a decrease in blood cells expressing adhesive (CD11b and CD62L) and functional (CD64 and HLA-DR) receptors. Moreover, the patient’s neutrophils expressed CD11b, CD16 and HLA-DR on their cell surface more intensively, than neutrophilic leukocytes from control group. These phenotypic changes in KC patients’ blood neutrophils occurred in parallel with pronounced decrease in immature cells numbers. The metabolic changes of neutrophil cytoplasmic compartment in KC patients were determined by a decrease in Glu6PDH activity (a key and initializing enzyme of the pentose phosphate cycle) and NADH-LDH (anaerobic glycolysis). Mitochondrial metabolism in neutrophils of KC patients was characterized by multidirectional changes in the activity of NAD- and NADP-dependent glutamate dehydrogenases (decreased activity of NAD-dependent and increased activity of NADP-dependent) and a decrease in NADH-MDH activity. The established features in mitochondrial enzymes activities suggest some disturbances of NAD-dependent processes that could lead to down-regulation of aerobic energy processes. We guess that the decreased activity of plastic and energy processes in blood neutrophils of KC patients could affect the receptor expression levels. By means of correlation analysis, we have found that the relationships in KC patients were determined by negative effects of NADHGDH and NADH-LDH activities upon expression of activation and adhesion receptors in blood neutrophils. Of these enzymes, only glutathione reductase activity in neutrophils from KC patients was positively linked with the CD23 and HLA-DR expression. Thus, an increase in activity of energy processes (e.g., coupling the tricarboxylic acid cycle to amino acid metabolism) in blood neutrophils from the patients with kidney cancer could stimulate expression levels of activation and adhesion receptors and potentially increase antitumor activity of neutrophils.
{"title":"Immunophenotype and metabolism are linked in peripheral blood neutrophils from patients with kidney cancer","authors":"A. Savchenko, A. Borisov, I. Kudryavtsev, I. Gvozdev, A. Moshev","doi":"10.15789/1563-0625-iam-2037","DOIUrl":"https://doi.org/10.15789/1563-0625-iam-2037","url":null,"abstract":"The aim of the present study was to analyze the relationships between expression of activation and adhesion receptors on peripheral blood neutrophils, and intracellular activity of some neutrophil enzymes in patients with kidney cancer (KC). Patients and methods: the KC patients (n = 72) (T3N0M0, clear-cell type) were examined prior to surgical treatment at the Krasnoyarsk Regional Oncology Center. The diagnosis was verified histologically for all KC patients. The phenotype of blood neutrophils was studied using flow cytometry. The surface receptor expression levels of the neutrophils were evaluated by mean fluorescence intensity. NAD and NADP-dependent dehydrogenases activities in purified peripheral blood neutrophils were measured by bioluminescent method. Results: we have found that the phenotypic alterations in circulating KC patients’ neutrophils appeared along with inhibition of main intracellular metabolic processes and were closely linked with them. The features of the phenotypic imbalance in the neutrophils from KC patients were associated with a decrease in blood cells expressing adhesive (CD11b and CD62L) and functional (CD64 and HLA-DR) receptors. Moreover, the patient’s neutrophils expressed CD11b, CD16 and HLA-DR on their cell surface more intensively, than neutrophilic leukocytes from control group. These phenotypic changes in KC patients’ blood neutrophils occurred in parallel with pronounced decrease in immature cells numbers. The metabolic changes of neutrophil cytoplasmic compartment in KC patients were determined by a decrease in Glu6PDH activity (a key and initializing enzyme of the pentose phosphate cycle) and NADH-LDH (anaerobic glycolysis). Mitochondrial metabolism in neutrophils of KC patients was characterized by multidirectional changes in the activity of NAD- and NADP-dependent glutamate dehydrogenases (decreased activity of NAD-dependent and increased activity of NADP-dependent) and a decrease in NADH-MDH activity. The established features in mitochondrial enzymes activities suggest some disturbances of NAD-dependent processes that could lead to down-regulation of aerobic energy processes. We guess that the decreased activity of plastic and energy processes in blood neutrophils of KC patients could affect the receptor expression levels. By means of correlation analysis, we have found that the relationships in KC patients were determined by negative effects of NADHGDH and NADH-LDH activities upon expression of activation and adhesion receptors in blood neutrophils. Of these enzymes, only glutathione reductase activity in neutrophils from KC patients was positively linked with the CD23 and HLA-DR expression. Thus, an increase in activity of energy processes (e.g., coupling the tricarboxylic acid cycle to amino acid metabolism) in blood neutrophils from the patients with kidney cancer could stimulate expression levels of activation and adhesion receptors and potentially increase antitumor activity of neutrophils.","PeriodicalId":85139,"journal":{"name":"Medical immunology (London, England)","volume":"22 1","pages":"887-896"},"PeriodicalIF":0.0,"publicationDate":"2020-11-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47181733","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}