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 responses of regulatory T cells induced by Sepharose 4B beads mixed antigen. Methods Mice were immunized with different dosages of antigens, and the proportions of regulatory T cells were examined by flow cytometry. The 3H-thymidin incorporation method was used to detect the inhibitory function of Tregs. In vitro, dendritic cells were pulsed with different antigens. The expressions of MHC II, CD40, CD80/86 molecules on dendritic cells and the proportions of regulatory T cells were examined by flow cytometry. The ^3H-thymidin incorporation method was used for the determination of Tregs function. Results Compared with control groups, the proportion of regulatory T cells in mice immunized with Sepharose 4B beads mixed antigen was(11.48±4.12)%, and the difference was significant (P〈0.05). The inhibition of regulatory T cells in mice immunized with Sepharose 4B beads mixed antigen showed a stronger potential (cpm was 720±180.4). Meanwhile, the proportions of regulatory T cells were (17.0±80.57)% and (30.14±3.62)% when the CD4^+ T cells were co-cultured with dendritic cells pulsed with Sepharose 4B beads mixed rSj22.6/26GST and OVA, respectively. Conclusion Sepharose 4B beads mixed antigen could induce the regulatory T cells in vivo and in vitro.