中文摘要：

目的 设计并鉴定胰岛素A链拮抗性HLA-A＊0201限制性CTL表位mInsA2-10或其APLs,为T1D防治的临床转化基础研究提供良好的实验依据。方法 腹腔注射可溶性肽mInsA2-10与无关对照肽,通过血糖监测判断肽mInsA2-10是否有T1D的防治作用;通过Insight II和Discover软件获得mInsA2-10和其APLs的分子动力学模拟参数,选择候选的APLs,再通过检测它们的相对亲和力及各候选APLs体外抑制mInsA2-10自身反应性CD8＋T细胞的增殖情况,初步鉴定具有拮抗作用的APLs,再系统性给予人源化NOD小鼠拮抗性APLs,考察其是否有T1D的防治作用。结果 比较分析mInsA2-10和其APLs的分子动力学模拟参数的差异,选择出4种候选的APLs;体外实验鉴定出APLs mInsA2-10DQ4和mInsA2-10DC6与HLA-A＊0201分子的相对亲和力比其它2个候选的APLs相对更高;与天然肽组相比,这2个APLs能显著地抑制mInsA2-10自身反应性的CD8＋T细胞的增殖;系统性给予人源化NOD小鼠这2个拮抗性APLs,仅mInsA2-10DQ4具有显著的T1D保护作用。结论 mInsA2-10DQ4和mInsA2-10DC6是具有拮抗作用的APLs,而在人源化NOD小鼠的在体研究中发现仅mInsA2-10DQ4具有显著的T1D保护作用,下一步将探讨mInsA2-10DQ4防治T1D的免疫学机制,这为T1D防治的临床转化基础研究提供良好的实验依据。

英文摘要：

To prevent the development of type 1 diabetes （T1D） by inducing peptide-specific autoreactive CD8^＋ T cell tolerance in humanized non-obese diabetic （NOD） mice, we designed and identified soluble HLA-A＊ 0201-restricted cytotoxic T lymphocyte （CTL） epitope mInsA2-10 and altered peptide ligands （APLs） derived from insulin A chain. Four APLs were designed and selected by molecular dynamics simulation in Insight Ⅱ and Discover software. The four altered peptide ligands （APLs） and mInsA2-10 were synthesized and the binding affinity of these APL candidates and native peptide for HLA-A＊0201 molecule was evaluated. Compared with mlnsA2-10DD3 and mInsA〉2-10DD3-DC6, mInsA2-10DQ4 and mInsA2-10DC6 had relatively higher binding affinity to HLA-A＊0201 molecule. Furthermore, mInsA2-10DQ4 and mInsA2-10DC6 significantly inhibited the proliferation of mInsA2-10 autoreaetive CD8^＋ T cells, as compared with mInsA2-10. The results demonstrated that mInsA2-10DQ4 and mInsA2-10DC6 are antagonistic APLs in vitro. And only mInsA2-10DQ4 is identified as a therapeutic candidate for in vivo studies, which would provide favorable experiment data for basic studies of clinical transformation.