Study on abnormalities of peripheral tolerance induction by target cells in organ-specific autoimmune diseases.

我们检测了自身免疫性甲状腺疾病患者甲状腺内淋巴细胞的凋亡和免疫偏差。TUNEL法检测DNA片段，双苯并胺H33258染色，7-氨基放线菌素d染色，证实甲状腺内淋巴细胞凋亡。免疫组化染色，检测甲状腺内淋巴细胞Fas分子表达，发现自身免疫性甲状腺疾病患者甲状腺内淋巴细胞Fas分子表达增加。此外，我们通过流式细胞仪分析CD4或CD8细胞与细胞质细胞因子（如IFNg、IL-2、IL-4、IL-5或IL-10）之间的不平衡，研究了Th1和Th2细胞数量之间以及Tc1和Tc2细胞数量之间的不平衡。然而，我们在自身免疫性甲状腺疾病中没有发现显著的结果，因为患者之间的差异很大，而且检查的患者数量很少。由于淋巴细胞与表达MHCⅱ类分子的甲状腺细胞共培养后淋巴细胞的恢复较小，因此未能有效检测甲状腺细胞中MHCⅱ类表达对淋巴细胞凋亡和免疫偏差的影响。

We examined apoptosis and immune deviation in intrathyroidal lymphocytes from patients with autoimmune thyroid disease. Apoptosis of intrathyroidal lymphocytes was confirmed by examining the DNA fragmentation by TUNEL method, the chromatin condensation and fragmentation by chromatin staining with bisbenzimide H33258, and the abnormal integrity of cell membrane by 7-amino-actinomycin D.The expression of Fas molecules on intrathyroidal lymphocytes was also examined by immunohistochemical staining, and found to be increased in autoimmune thyroid disease. Furthermore, we examined imbalance between the numbers of Th1 and Th2 cells and between those of Tc1 and Tc2 cells by analyzing CD4 or CD8 cells with cytoplasmic cytokines such as IFNg, IL-2, IL-4, IL-5, or IL-10, with a flow cytometer. However, we found no significant results in autoimmune thyroid disease, because of large variation among patients and of small number of patients examined. The effect of MHC class II expression on thyrocytes on apoptosis and immune deviation of lymphocytes was not efficiently examined because of small recovery of lymphocytes after co-culture of lymphocytes and thyrocytes expressing MHC class II molecules.