Luo Xue-Ping, Chen Dian-Hui, Xie Hong-Yan, Gao Zhi-Yan, Fang Hui-Long, Huang Jun
Department of Pathogenic Biology and Immunology, Guangzhou Medical College, Guangzhou 510182, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2012 Aug 30;30(4):258-61, 267.
To observe the immune response of Th17 cells in mesenteric lymph node (MLN) of C57BL/6 mice infected by Schistosoma japonicum.
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.
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.
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.
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。
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。
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。
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。
