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目的分析小鼠脾细胞中CD4+CD25+Foxp3+T细胞在日本血吸虫感染后的动态变化,探讨CD4+CD25+Foxp3+T细胞与血吸虫感染的关系。方法常规逸出尾蚴,感染小鼠24只,随机分为4组,同时设立阴性对照。感染后第3、6、9、12周分批处死小鼠,取脾脏制备单细胞悬液,用FITC-CD4和PE-CD25单抗进行细胞表面染色,洗涤后,经打孔液和固定液处理,再用PE-cy5-Foxp3抗体进行核内染色,流式细胞仪检测CD4+CD25+Foxp3+调节性T细胞的变化。结果小鼠感染血吸虫后第3、6、9、12周脾细胞中CD4+CD25+Foxp3+T细胞百分率分别为(2.57±0.13)%、(1.77±0.21)%、(1.10±0.05)%和(0.83±0.04)%,呈逐步下降趋势(P<0.05);CD4+CD25-Foxp3+T细胞百分率分别为(0.25±0.007)%、(0.27±0.13)%、(0.38±0.15)%和(0.35±0.07)%,差异无统计学意义(P>0.05)。未感染小鼠脾细胞中CD4+CD25+Foxp3+T细胞百分率分别为(2.63±0.09)%、(3.13±0.47)%、(2.78±0.19)%和(2.82±0.12)%,差异无统计学意义(P>0.05);CD4+CD25-Foxp3+T细胞百分率分别为(0.35±0.10)%、(0.43±0.13)%、(0.30±0.08)%和(0.31±0.09)%,差异无统计学意义(P>0.05)。感染组与未感染组相比从第6周开始脾细胞中CD4+CD25+Foxp3+T细胞百分率显著下降(P<0.05),CD4+CD25-Foxp3+T细胞百分率差异无统计学意义(P>0.05)。结论CD4+CD25+Foxp3+T细胞在日本血吸虫感染中具有免疫抑制作用,与血吸虫虫卵肉芽肿慢性病变具有相关性。
Objective To analyze the dynamic changes of CD4 + CD25 + Foxp3 + T cells in mice spleen cells after infection with Schistosoma japonicum and explore the relationship between CD4 + CD25 + Foxp3 + T cells and schistosomiasis infection. Methods Totally 24 cercariae were infected with cercariae and were randomly divided into 4 groups. At the same time, a negative control was established. Mice were sacrificed in batches at 3, 6, 9 and 12 weeks after infection. Single spleen suspensions were prepared and stained with FITC-CD4 and PE-CD25 monoclonal antibodies. After washing, The cells were stained with PE-cy5-Foxp3 antibody, and the changes of CD4 + CD25 + Foxp3 + regulatory T cells were detected by flow cytometry. Results The percentage of CD4 + CD25 + Foxp3 + T cells in splenocytes of mice infected with Schistosoma japonicum at the 3rd, 6th, 9th and 12th week were (2.57 ± 0.13)%, (1.77 ± 0.21)%, (1.10 ± 0.05)% and (0.83 ± 0.04)%, and the percentage of CD4 + 0.35 ± 0.07)%, the difference was not statistically significant (P> 0.05). The percentages of CD4 + CD25 + Foxp3 + T cells in uninfected mice were (2.63 ± 0.09)%, (3.13 ± 0.47)%, (2.78 ± 0.19)% and (2.82 ± 0.12)%, respectively (P <0.05). The percentages of CD4 + CD25-Foxp3 + T cells were (0.35 ± 0.10)%, (0.43 ± 0.13)%, (0.30 ± 0.08)% and Significance (P> 0.05). The percentage of CD4 + CD25 + Foxp3 + T cells in spleen cells decreased significantly (P <0.05) and the percentage of CD4 + CD25-Foxp3 + T cells in spleen cells from the 6th week in infection group was not significantly different (P> 0.05). Conclusion CD4 + CD25 + Foxp3 + T cells have immunosuppressive effect on Schistosoma japonicum infection, which is correlated with the chronic lesions of Schistosoma japonicum egg granuloma.