摘要
目的探讨多房棘球幼(E.m)感染所致的免疫应答偏倚对实验性类风湿性关节炎(CIA)的免疫调节作用。方法分别建立雌性C57小鼠胶原性关节炎模型(CIA组)、多房棘球蚴感染模型(E.m组)和多房棘球蚴感染后类风湿性关节炎诱导模型(E.m+CIA组),利用实时荧光定量RT-PCR和酶联免疫吸附试验(ELISA)检测小鼠足爪组织、外周血及脾脏细胞中Th17细胞相关因子及转录因子水平,实验设阴性对照组(Control组)。组间比较采用非参数秩和检验两个独立样本比较法(M-W法);采用Spearman线性相关性分析方法分析细胞因子及转录因子之间的相关关系。结果外周血及脾脏细胞中IL-17A、IL-1β、IFN-γ水平在E.m+CIA组18.93(14.90-20.53)pg/ml、18.93(14.90-20.53)pg/ml,4.18(1.79-4.45)pg/ml、4.182(1.794-4.446)pg/ml,14.79(0.64-29.38)pg/ml、14.79(0.64-29.38)pg/ml,显著低于CIA组16.47(15.41-19.46)pg/ml、16.47(15.41-19.46)pg/ml,161.60(125.40-222.20)pg/ml、367.2(240.4-550.3)pg/ml,29.89(19.93-58.95)pg/ml、29.89(19.93-58.95)pg/ml(P〈0.05或P〈0.01);E.m+CIA组IL-4、IL-6等抗炎相关因子水平11.68(11.00-12.85)pg/ml、11.68(11.00-12.85)pg/ml,450.40(292.80-697.50)pg/ml、450.40(292.80-645.60)pg/ml,较其他组显著升高(P〈0.05或P〈0.01)。小鼠足爪中IL-17A及RORrt mRNA相对表达量在E.m+CIA组13.29(5.38-18.93),1.07(0.78-6.99)较其他组降低。结论 E.m感染诱导的Th2优势应答可使CIA不能有效启动致病性Th1免疫应答,抑制CIA小鼠过度的Th17应答,从而减轻RA炎症损伤。
Objective To explore the effects of immune response bias caused by infection with Echinococcus multilocularis on immune system regulation as a result of experimentally induced rheumatoid arthritis. Methods Thirty-two 5-6week old C57 mice were randomly divided into four groups(n=8in each group):(1)negative control group(control),(2)E.multilocularis group:healthy mice were inoculated with E.multilocularis,(3)CIA group:rheumatoid arthritis was induced in healthy mice using collagen,(4)E.multilocularis+CIA group:healthy mice were infected with E.multilocularis and then rheumatoid arthritis was induced with collagen.The levels of Th17 cells and Th17-related cytokines in the spleen and peripheral blood and the tissue of the paw were detected using RT-PCR and ELISA.Groups were compared using a non-parametric rank sum test for two independent samples.Spearman linear correlation analysis was used to study the relationship between cytokines and transcription factors. Results In the E.multilocularis+CIA group,the level of IL-17 Ain peripheral blood was 18.93pg/ml(range:14.90-20.53pg/ml)and that in spleen cells was 18.93pg/ml(range:14.90-20.53pg/ml),the level of IL-1βin peripheral blood was 4.18pg/ml(range:1.79-4.45pg/ml)and that in spleen cells was 4.182pg/ml(range:1.794-4.446pg/ml),and the level of IFN-γin peripheral blood was14.79pg/ml(range:0.64-29.38pg/ml)and that in spleen cells was 14.79pg/ml(range:0.64-29.38pg/ml).In the CIA group,the level of IL-17 Ain peripheral blood was 16.47pg/ml(range:15.41-19.46pg/ml)and that in spleen cells was 16.47pg/ml(range:15.41-19.46pg/ml),the level of IL-1βin peripheral blood was 161.60pg/ml(range:125.40-222.20pg/ml)and that in spleen cells was 367.2pg/ml(range:240.4-550.3pg/ml),and the level of IFN-γin peripheral blood was 29.89pg/ml(range:19.93-58.95pg/ml)and that in spleen cells was 29.89pg/ml(range:19.93-58.95pg/ml).The levels of IL-17 A,IL-1β,and IFN-γin peripheral blood and spleen cells were lower in the E.multilocularis+CIA group than those in the CIA group.In the E.multilocularis+CIA group,the level of the anti-inflammatory cytokine IL-4in peripheral blood was 11.68pg/ml(range:11.00-12.85pg/ml)and that in spleen cells was11.68pg/ml(range:11.00-12.85pg/ml)and the level of the anti-inflammatory cytokine IL-6in peripheral blood was450.40pg/ml(range:292.80-697.50pg/ml)and that in spleen cells was 450.40pg/ml(range:292.80-645.60pg/ml).The levels of anti-inflammatory cytokines like IL-4and IL-6were higher in the E.multilocularis+CIA group than those in the CIA group.The level of IL-17 A mRNA expression in the tissue of the paw was 23.94(range:17.04-39.50)in the CIA group and 13.29(range:5.38-18.93)in the E.multilocularis+CIA group,so the level of IL-17 A mRNA expression decreased significantly(P〈0.05).The level of RORrt mRNA expression was 1.07(range:0.78-6.99)in the E.multilocularis+CIA group and 1.56(range:1.23-6.71)in the CIA group,so the level of RORrt mRNA expression did not differ significantly between the two groups(P〉0.05). Conclusion Infection with E.multilocularis induced a Th2-dominant immune response but CIA precluded the effective initiation of a pathogenic Th1 immune response.In addition,infection with E.multilocularis markedly inhibited an excessive Th17 immune response in mice with CIA.These two findings will help to reduce the inflammatory damage of rheumatoid arthritis.
出处
《中国病原生物学杂志》
CSCD
北大核心
2016年第10期917-923,共7页
Journal of Pathogen Biology
基金
新疆维吾尔自治区自然科学基金项目(No.2014211C081)