论文部分内容阅读
目的研究Sepharose 4B颗粒混合日本血吸虫22.6kDa/26GST抗原对CD4+CD25+调节性T细胞(Tregs)的诱导作用。方法将不同剂型抗原免疫小鼠,流式细胞术检测脾细胞中Tregs占CD4+T细胞的比例,3H-TDR掺入法检测Tregs的功能。体外用颗粒化抗原负载树突状细胞(DCs),流式细胞术检测DCs表面MHCII类分子、CD40、CD80、CD86,以判定DCs的成熟度,同时用流式细胞术检测负载抗原后的DCs对Tregs的诱导作用。结果Sepharose 4B颗粒混合日本血吸虫22.6kDa/26GST抗原免疫小鼠的脾细胞中,Tregs比例为(11.48±4.12)%,均较其他各对照组高(P均<0.05),其抑制功能(cpm值为720±180.4)亦较其他各对照组增强。体外将脾CD4+T细胞与负载过Sepharose4B颗粒混合rSj22.6/26GST抗原的DCs共培养后,Tregs所占比例为(17.08±80.57)%;而与Sepharose 4B颗粒混合卵白蛋白负载过的树突状细胞共培养后,Tregs所占比例为(30.14±3.62)%,均较对照组增高(P均<0.01)。结论 Sepharose 4B颗粒与抗原蛋白共同存在可以诱导调节性T细胞。
Objective To study the induction of CD4 + CD25 + regulatory T cells (Tregs) by Sepharose 4B mixed with 22.6kDa / 26GST antigen of Schistosoma japonicum. Methods Different doses of antigens were used to immunize mice. Flow cytometry was used to detect the proportion of Tregs to CD4 + T cells in spleen cells. 3H-TDR incorporation assay was used to detect Tregs function. Dendritic cells (DCs) were loaded with granulocyte antigen in vitro. Flow cytometry was used to detect the MHC class II molecules, CD40, CD80 and CD86 on the surface of DCs to determine the maturation of DCs. Flow cytometry was used to detect DCs Induction of Tregs. Results The percentage of Tregs in spleen cells of mice immunized with Sepharose 4B particles and 22.6kDa / 26GST antigen of Schistosoma japonicum was (11.48 ± 4.12)% higher than that of other control groups (all P <0.05) 720 ± 180.4) than other control groups. In vitro co-culture of splenic CD4 + T cells with DCs loaded with Sepharose4B particles mixed with rSj22.6 / 26GST antigen, the percentage of Tregs was (17.08 ± 80.57)%, while in combination with Sepharose 4B particles, ovalbumin-loaded dendrites The percentage of Tregs was (30.14 ± 3.62)% in co-cultured cells compared with control group (all P <0.01). Conclusion Sepharose 4B particles coexist with antigenic proteins to induce regulatory T cells.