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日本血吸虫感染小鼠肠系膜淋巴结中Th17细胞的免疫应答

[Immune response of Th17 cells in mesenteric lymph node of mice infected by Schistosoma japonicum].

作者信息

Luo Xue-Ping, Chen Dian-Hui, Xie Hong-Yan, Gao Zhi-Yan, Fang Hui-Long, Huang Jun

机构信息

Department of Pathogenic Biology and Immunology, Guangzhou Medical College, Guangzhou 510182, China.

出版信息

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2012 Aug 30;30(4):258-61, 267.

Abstract

OBJECTIVE

To observe the immune response of Th17 cells in mesenteric lymph node (MLN) of C57BL/6 mice infected by Schistosoma japonicum.

METHODS

Twenty C57BL/6 mice were randomly divided into infected group and control group each with ten mice. The mice in infected group were infected each with 40 +/- 5 S. japonicum cercariae. Five to six weeks later, MLN lymphocytes were separated and stimulated for 4 h by anti-CD3 (1 microg/ml) and anti-CD28 (1 microg/ml) before examination of IL-17 and retinoic acid receptor-related orphan receptor gammat (ROR-gammat) mRNA by reverse transcription PCR. The level of IL-17 and IFN-gamma was detected by ELISA after culturing with supernatant for 72h. MLN lymphocytes were stimulated for 5h by 10 ng/ml phorbol myristoyl acetate (PMA) and 1 microg/ml ionomycin. The intracellular cytokines were stained and the content of Th17 and other cytokines was examined by flow cytometry.

RESULTS

The level of IFN-gamma [(214.3 +/- 62.6) pg/ml] and IL-17 [(176.8 +/- 62.1) pg/ml] in the supernatant of cultured MLN cells from the infected mice was significantly higher than that of normal mice [(467 +/- 13.9) and 0 pg/ml) (P < 0.05). The expression level of IL-17 and ROR-gammat mRNA was also considerably higher than that of normal mice. IL-17+ IL-4+, IL-17+ IFN-gamma+, IL-17+ IL-5+ and IL-17+ IL-9 cells accounted for 0.06%, 0.02%, 0.02%, and 0.01% of the mesenteric lymph node CD4+ T cells of the infected mice, respectively. However, IL-17+ IL-10+ and IL-17+ Foxp3+ cells were undetected.

CONCLUSION

The MLN of S. japonicum-infected C57BL/6 mice can induce the production of Th17 cells, and these cells can secrete IL-4, less IFN-gamma, IL-5 and IL-9, but not IL-10, and can not express Foxp3 in the infected mice.

摘要

目的

观察日本血吸虫感染的C57BL/6小鼠肠系膜淋巴结(MLN)中Th17细胞的免疫反应。

方法

将20只C57BL/6小鼠随机分为感染组和对照组,每组10只。感染组小鼠每只感染40±5条日本血吸虫尾蚴。5至6周后,分离MLN淋巴细胞,用抗CD3(1μg/ml)和抗CD28(1μg/ml)刺激4小时,然后通过逆转录PCR检测IL-17和视黄酸受体相关孤儿受体γt(ROR-γt)mRNA。用培养上清培养72小时后,通过ELISA检测IL-17和IFN-γ水平。用10ng/ml佛波酯(PMA)和1μg/ml离子霉素刺激MLN淋巴细胞5小时。对细胞内细胞因子进行染色,通过流式细胞术检测Th17和其他细胞因子的含量。

结果

感染小鼠培养的MLN细胞上清中IFN-γ水平[(214.3±62.6)pg/ml]和IL-17水平[(176.8±62.1)pg/ml]显著高于正常小鼠[(467±13.9)和0pg/ml](P<0.05)。IL-17和ROR-γt mRNA的表达水平也明显高于正常小鼠。IL-17+IL-4+、IL-17+IFN-γ+、IL-17+IL-5+和IL-17+IL-9细胞分别占感染小鼠肠系膜淋巴结CD4+T细胞的0.06%、0.02%、0.02%和0.01%。然而,未检测到IL-17+IL-10+和IL-17+Foxp3+细胞。

结论

日本血吸虫感染的C57BL/6小鼠的MLN可诱导Th17细胞产生,这些细胞可分泌IL-4、较少的IFN-γ、IL-5和IL-9,但不分泌IL-10,且在感染小鼠中不表达Foxp3。

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