约氏疟原虫感染小鼠树突状细胞对Th17分化和功能的调控作用

发布时间：2018-04-17 05:14

本文选题：约氏疟原虫 + 树突状细胞　；参考：《中国寄生虫学与寄生虫病杂志》2017年01期

【摘要】：目的探讨约氏疟原虫(Plasmodium yoelii)17XL株(Py17XL)感染小鼠过程中树突状细胞(DCs)对辅助性T细胞17型(Thl7)细胞分化和功能的调控作用及机制。方法将12只雌性BALB/c小鼠随机分为感染组(Py17XL)、Toll样受体4(TLR4)阻断组(Py17XL+TLR4)、Toll样受体9(TLR9)阻断组(Py17XL+TLR9)和TLR4联合TLR9阻断组(Py17XL+TLR4+TLR9),每组3只。感染前1 d,腹腔注射10μg抗-TLR4单抗(TLR4阻断组)或50μg抗-TLR9单抗(TLR9阻断组)阻断DCs的功能,注射体积为0.4 ml,对照组注射等量的PBS。4组小鼠均腹腔注射1×10~6 Py17XL感染的红细胞。感染后第0、3和5天计数红细胞感染率,同时制备脾细胞悬液,流式细胞仪检测脾脏细胞中CD11c+TLR9+和Th17细胞数量变化,ELISA检测脾细胞培养上清中干扰素-γ(IFN-γ)和白细胞介素-10(IL-10)水平的变化。结果流式细胞仪检测表明,DCs阻断模型构建成功。感染后第5天,感染组、TLR4阻断组、TLR9阻断组和TLR4联合TLR9阻断组小鼠原虫血症分别为28.0%、29.0%、31.0%和16.3%;感染后第7天,原虫血症分别为43.3%、47.5%、32.5%和8.0%,感染组和TLR4阻断组小鼠全部死亡,其余阻断组小鼠于感染后第6天开始出现死亡。与感染组比较,TLR9阻断组和TLR4联合TLR9阻断组小鼠原虫血症上升缓慢,生存期明显延长至11 d和15 d。流式细胞仪检测结果显示:感染后第5天,TLR9阻断组和TLR4联合TLR9阻断组Th17细胞数量分别为1.2%和1.4%,与感染组(1.9%)相比,数量明显减少(P0.05,P0.01)。ELISA检测结果显示:感染后第5天,TLR4联合TLR9阻断组小鼠IFN-γ和IL-10水平分别为(232.4±15.5)pg/ml和(1 791.2±58.2)pg/ml,与感染组[(90.7±50.1)pg/ml和(962.6±409.0)pg/ml]相比,差异有统计学意义(P0.05,P0.01)。结论 Py17XL感染小鼠过程中DCs调控了Thl7细胞的分化和功能。DCs阻断(Py17XL+TLR4+TLR9)后,感染小鼠原虫血症降低,生存期延长。
[Abstract]:Objective to investigate the effect and mechanism of dendritic cells (DCs) on the differentiation and function of helper T cell type 17 (Thl7) cells in mice infected with Plasmodium yoelii)17XL strain Py17 XLL.Methods Twelve female BALB/c mice were randomly divided into infected group (Py17XL TLR4) and TLR4 combined with TLR9 (3 mice in each group).One day before infection, 10 渭 g anti-TLR4 monoclonal antibody (10 渭 g TLR4) or 50 渭 g anti-TLR9 monoclonal antibody (50 渭 g anti-TLR9) blocked the function of DCs. The volume of PBS.4 was 0.4 ml. In control group, the same amount of PBS.4 mice were injected intraperitoneally with red blood cells infected with 1 脳 10 6 Py17XL.The infection rate of erythrocyte was counted on the 3rd and 5th day after infection, and spleen cell suspension was prepared. The changes of CD11c TLR9 and Th17 cells in spleen cell were detected by flow cytometry. The levels of IFN- 纬 and IL-10 in the supernatant of spleen cell culture were detected by Elisa.Results flow cytometry showed that the blocking model of DCs was successfully constructed.On the 5th day after infection, the protozoaemia of the infected mice in the TLR9 blocking group and the TLR4 combined TLR9 blocking group were 28.00.29.0% and 16.30.On the 7th day after infection, the rates of protozoemia were 43.33.5% and 8.0%, respectively. All the mice in the infection group and TLR4 block group died.The rest of the mice in the blocking group began to die on the 6th day after infection.Compared with the infected group, the parasitemia of mice in the TLR9 group and the TLR4 combined with TLR9 group increased slowly, and the survival time was prolonged to 11 days and 15 days.The results of flow cytometry showed that the number of Th17 cells in the TLR9 blocking group and the TLR4 combined TLR9 blocking group were 1.2% and 1.4 on the 5th day after infection, respectively, compared with that in the infected group (1.9%).The results of Elisa showed that the levels of IFN- 纬 and IL-10 were 232.4 卤15.5)pg/ml and 1 791.2 卤58.2 渭 g / ml in the TLR4 + TLR9 group on the 5th day after infection, which were significantly different from those in the infected group [90.7 卤50.1)pg/ml and 962.6 卤409.0)pg/ml].Conclusion during the course of Py17XL infection, DCs regulates the differentiation and function of Thl7 cells. After blocking Py17XL TLR4 TLR9, the parasitemia of infected mice decreases and the survival time is prolonged.
【作者单位】：佳木斯大学基础医学院;佳木斯大学附属第一医院;
【基金】：黑龙江省教育厅海外学人资助项目(No.1254HQ017)~~
【分类号】：R382.31

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