Bedin C, Brazillet M P, Texier B, Charreire J
INSERM U 283, Hôpital COCHIN, Paris.
Int Rev Immunol. 1992;9(2):125-33. doi: 10.3109/08830189209061787.
These data collect the advance made in the last few years in our laboratory in defining one epitope from the thyroglobulin (Tg) molecule (660 KDa) inducing Experimental Autoimmune thyroiditis (EAT) in CBA/J mice. We achieved the characterization of one EAT-inducer Tg peptide by combining "in vitro" biochemical and immunological approaches and "in vivo" studies. Since T cells recognize degraded forms of the antigen and since endogenous antigens preferentially activate class I-restricted T cells, we hypothesized that one cytotoxic T cell hybridoma, named HTC2, which prevents further EAT induction in mice injected with Tg would be specific for one EAT inducer peptide. In order to identify one Tg epitope inducing EAT, enzymatic treatment of the protein by trypsin, HPLC purification and sequence analysis were performed. Simultaneously, tryptic digests were used to pulse CBA/J macrophages and tested for their ability to be recognized by HTC2 cells. Lastly, when digests were recognized by HTC2 cells their capacity to induce EAT in CBA/J mice was evaluated. To further assess the pathogenicity of the sequenced Tg peptide, one synthetic peptide was made and its capacity to induce EAT verified. By this procedure we identified for the first time one 40 amino-acid peptide from human thyroglobulin inducing EAT in CBA/J mice.
这些数据收集了我们实验室在过去几年中在确定甲状腺球蛋白（Tg，660 kDa）分子中的一个表位方面取得的进展，该表位可在CBA/J小鼠中诱发实验性自身免疫性甲状腺炎（EAT）。我们通过结合“体外”生化和免疫学方法以及“体内”研究，实现了对一种EAT诱导性Tg肽的表征。由于T细胞识别抗原的降解形式，并且内源性抗原优先激活I类限制性T细胞，我们假设一种名为HTC2的细胞毒性T细胞杂交瘤，它能阻止注射Tg的小鼠进一步发生EAT诱导，将对一种EAT诱导肽具有特异性。为了鉴定一种诱导EAT的Tg表位，我们对该蛋白质进行了胰蛋白酶酶解处理、HPLC纯化和序列分析。同时，用胰蛋白酶消化产物脉冲CBA/J巨噬细胞，并测试它们被HTC2细胞识别的能力。最后，当消化产物被HTC2细胞识别时，评估它们在CBA/J小鼠中诱导EAT的能力。为了进一步评估测序的Tg肽的致病性，合成了一种合成肽并验证了其诱导EAT的能力。通过这个程序，我们首次从人甲状腺球蛋白中鉴定出一种40个氨基酸的肽，它能在CBA/J小鼠中诱导EAT。
