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离。甲状腺内淋巴细胞的凋亡通过TUNEL法检测DNA片段化、双苯甲酰亚胺H33258染色质染色检测染色质凝聚和片段化、7-氨基-放线菌素D染色检测细胞膜异常完整性来证实。免疫组化染色检测甲状腺内淋巴细胞Fas分子的表达，发现自身免疫性甲状腺疾病中Fas分子的表达增加。此外，我们通过用流式细胞仪分析具有细胞质细胞因子如IFNg、IL-2、IL-4、IL-5或IL-10的CD 4或CD 8细胞，来检查Th 1和Th 2细胞的数量之间以及Tc 1和Tc 2细胞的数量之间的不平衡。然而，我们没有发现显着的结果在自身免疫性甲状腺疾病，因为患者之间的差异大，检查的患者人数少。由于表达MHC II类分子的甲状腺细胞和淋巴细胞共培养后淋巴细胞的恢复率很小，因此没有有效地检测到MHC II类分子对甲状腺细胞的表达对淋巴细胞凋亡和免疫偏离的影响。

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.