摘要
目的:探讨PI-IBS小鼠γδT细胞的表型和功能及其在PI-IBS发病中的作用。方法:旋毛虫感染小鼠,观察肠道炎症、腹壁撤退反射(内脏高敏感性)和结肠传输时间(肠道动力)。分别在感染后第2周和第8周处死动物,取末端回肠和近端结肠组织,免疫荧光组化,激光扫描共聚焦显微镜观察肠黏膜γδT细胞的分布和数量变化。收集PI-IBS小鼠肠道淋巴结和脾脏的淋巴细胞,单克隆抗体免疫磁珠分选法分离和纯化γδT细胞,3HTdR法检测其增殖情况,FACS检测其表面分子CD69、CD62L,ELISA检测细胞培养上清液中IFN-γ及IL-17的表达。结果:感染后第2周,PI-IBS小鼠肠道炎症明显,肠黏膜γδT细胞数量明显增加,明显增殖和活化,产生IL-17明显增加(P<0.01)。感染后第8周,PI-IBS小鼠肠道炎症基本消退,动物的腹壁撤退反射和结肠传输试验明显异常。肠黏膜γδT细胞数量仍然明显高于对照组小鼠,仍然有明显增殖、活化和产生IL-17(P<0.01)。结论:肠道γδT细胞增殖、活化及分泌IL-17可能参与PI-IBS的发病。
Objective： To investigate the phenotype and function of the intestinal γδT lymphocytes in post-infectious irritable bowel syndrome mouse model. Methods： The mouse model for post-infectious irritable bowel syndrome was established by the infection with trichinella spiralis. The intestinal inflammation,abdominal withdrawal reflex（ AWR） and colon transportation test were observed. 2and 8 weeks later,the animals were sacrificed and the lymphocytes in the intestinal lymph nodes and spleen were collected,from which the γδT lymphocytes were isolated and purified by monoclonal antibody-immuno-microbeads method. The functions of the purified γδT lymphocytes were evaluated,including proliferation by^3HTdR; CD69,CD62 L molecule staining by flow cytometry. Furthermore,the concentration of cytokine IL-17 and IFN-γ in the supernatant of the cultured γδT lymphocytes were detected by ELISA. Results： At 2nd weeks after infection,significant intestinal inflammation was observed,with increasing γδT lymphocytes,proliferating and activating with increasing production of IL-17. At 8th weeks after infection,the intestinal inflammation disappeared,whereas the number of γδT lymphocytes remained increasing,also with proliferating and activating with increasing production of IL-17. Meanwhile,the mice show higher AWR score and Bristol score. Conclusion： γδT lymphocytes could participate in the pathogenesis of PI-IBS via their proliferation,activation and production of IL-17.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2016年第7期957-961,共5页
Chinese Journal of Immunology
基金
国家自然科学基金(No.81160057)
海南省国际科技合作专项基金(No.KJHZ2013-14)
关键词
感染后肠易激综合征
ΓΔT细胞
炎症
表型
功能
Post-infectious irritable bowel syndrome
γδT cells
Inflammation
Phenotype
Function
