CD4^+ FoxP3^+ Treg细胞在幽门螺杆菌感染C57BL/6小鼠中作用的初步研究

Primary study of role of CD4^+ FoxP3^+ Treg cells in C57BL /6 mice infected with Helicobacter pylori

目的:探索CD4+FoxP3+Treg细胞在幽门螺杆菌(Helicobbacter pylori,H.pylori)感染中作用。方法:建立H.pylori感染的小鼠模型,同时设立对照组。流式细胞仪检测胃黏膜固有层淋巴细胞中CD4+FoxP3+Treg细胞的比率;RTPCR检测胃组织中foxp3基因的表达;ELISA检测血清中IL-10、TGF-β1、IFN-γ的水平。用H.pylori全菌超声抗原刺激感染组脾淋巴细胞,流式细胞术检测Treg细胞的变化。ELISA检测相关细胞因子的水平。结果:感染组小鼠胃黏膜固有层淋巴细胞中CD4+FoxP3+Treg细胞的比率升高(P<0.01),胃组织高表达foxp3基因。血清中IFN-γ含量增高(P<0.05)。脾淋巴细胞经抗原刺激后,H.pylori抗原刺激组和阳性刺激组中CD4+FoxP3+Treg细胞比率上升(P<0.01),且大量分泌抑制性细胞因子IL-10(P<0.05)和TGF-β1(P<0.05)结论:CD4+Foxp3+Treg细胞与H.pylori感染密切相关。

Objective：To investigate the role of CD4 ＋ FoxP3 ＋ Treg cells in the H. pylori infection. Methods： To establish a mouse model of H. pylori infectous gastritis, meanwhile control group were set up. Percentage of CD4 ＋ FoxP3 ＋ Treg cells in LPLs （ lamina proprial lymphocytes, LPLs）was analysed by Flow cytometry. RT-PCR was used to detect the expression offoxp3 gene. The level of IL- 10,TGF-β1 ,IFN-γ, in serum were determined by ELISA. Spleen lymphocytes from infected group cultured and stimulated by H. pylori antigen was analysed by Flow cytometry, the cytokines in the supematant were detected by ELISA. Results： Compare with control group ,Percentage of CD4＋FoxP3＋Treg cells in LPLs increased significantly（ P〈0. 01 ） in infection group mouse, foxp3 gene enhanced expressed in gastric tissue. IFN-γ in serum was higher than that of PBS group（P〈0.05）. After antigen stimulation, more spleen T lym- phocytes transformed into CD4＋FoxP3＋Treg cells（ P〈0.01 ） in H. pylori antigen group and positive group and secreted much more IL-10 （P〈0. 01 ） and TGF-β1 （P〈O. 05 ）. Conclusion： CD4＋FoxP3＋Treg cells is closely related to H. pylori infection in C57BL/6 mice.