目的观察分析人源化NOD小鼠体内CD8^+CD25^+T细胞的频率及功能,探讨CD8^+CD25^+T细胞在人源化NOD小鼠的自身免疫进程中的作用。方法采用流式细胞术检测人源化NOD小鼠脾、胰腺引流淋巴结、腹股沟淋巴结中CD8^+CD25^+T、CD8^+CD25^+Foxp3+T细胞的频率及脾CD8^+CD25^+T细胞的表型和细胞因子分泌谱,并采用3H-Td R掺入法检测脾CD8^+CD25^+T细胞的免疫抑制功能。结果人源化NOD小鼠胰腺引流淋巴结中CD8^+CD25^+T和CD8^+CD25^+Foxp3+T细胞频率显著高于脾和腹股沟淋巴结中这2亚群的频率;与CD8^+CD25-T细胞相比,脾CD8^+CD25^+T细胞高表达Foxp3、CTLA-4、CD103、CD62L,同时低分泌细胞因子IFN-γ及高分泌IL-17A;人源化NOD小鼠脾CD8^+CD25^+T细胞能有效地抑制效应性CD4+CD25-T细胞的增殖。结论人源化NOD小鼠的CD8^+CD25^+T细胞是一群具有免疫抑制功能的Tregs,并且高表达Tregs的表型和细胞因子谱,提示CD8^+CD25^+Tregs可能在外周和胰腺局部参与了人源化NOD小鼠T1D的免疫耐受进程。
To evaluate the role of CD8^+CD25^+T cells in the process of autoimmunity of humanized NOD mice,the function and frequency changes of CD8^+CD25^+T cells in the process were observed. Flow cytometry wasemployed to analyze the CD8^+CD25^+T and CD8^+CD25^+Foxp3+T cell frequencies of pancreas drainage lymph gland(PDLN), inguinal lymph node(ILN), and spleen; differences of phenotype and cytokine profile between splenic CD8^+CD25^+and CD8^+CD25-T cells were also analyzed by flow cytometry. Data showed CD8^+CD25^+and CD8^+CD25^+Foxp3+T cell frequencies of PDLN were higher than those of ILN and spleen. Compared with CD8^+CD25-T cells, theCD8^+CD25^+T cell subpopulation harbored a typical Treg phenotype with high expression of Foxp3, CD103, CD62 Land CTLA-4, and inversely low level of IFN-γ and high level of IL-17 A. Furthermore, splenic CD8^+CD25^+T cellssignificantly inhibited the proliferation of CD4+CD25-T cells. These results demonstrated that splenic CD8^+CD25^+Tcells in humanized NOD mice expresse classical Treg phenotype and cytokine profile and has significantimmunosuppressive function, thus might play an important immunosuppressive role in autoimmunity process ofhumanized NOD mice.