Abstract:

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 cocultured 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.

Key words: Schistosoma japonicum, Sepharose 4B beads, Dendritic cells, Regulatory T cells

摘要：

目的 研究Sepharose 4B颗粒混合日本血吸虫22.6 kDa/26 GST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， 3 H?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MHC II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22.6 kDa/26 GST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Sepharose 4B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。

关键词: 日本血吸虫；Sepharose 4B；树突状细胞；调节性T细胞