论文部分内容阅读
目的设计并鉴定胰岛素A链拮抗性HLA-A*0201限制性CTL表位mInsA_(2-10)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mInsA_(2-10)与无关对照肽,通过血糖监测判断肽mInsA_(2-10)是否有T1D的防治作用;通过Insight II和Discover软件获得mInsA_(2-10)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mInsA_(2-10)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mInsA_(2-10)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 mInsA_(2-10)DQ4和mInsA_(2-10)DC6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mInsA_(2-10)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mInsA_(2-10)DQ4具有显著的T1D保护作用。结论 mInsA_(2-10)DQ4和mInsA_(2-10)DC6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mInsA_(2-10)DQ4具有显著的T1D保护作用,下一步将探讨mInsA_(2-10)DQ4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。
Objective To design and identify the mInsA_ (2-10) or its APLs of HLA-A * 0201-restricted CTL epitope of insulin A chain and provide a good experimental basis for the clinical research of T1D prevention and control. Methods The soluble mlsA (2-10) and the unrelated control peptide were injected intraperitoneally to determine whether the mInsA_ (2-10) had the preventive and therapeutic effects on T1D. The results of Insight II and Discover software showed that mInsA_ (2-10) and its APLs (APLs) were selected and their relative affinities were tested. The APLs with antagonistic effects were initially identified by detecting the proliferation of mInsA 2- (2-10) autoreactive CD8 + T cells in vitro. Antagonistic APLs were given to humanized NOD mice systematically to investigate whether they had the preventive and therapeutic effects on T1D. Results The molecular dynamics simulation parameters of mInsA (2-10) and its APLs were compared and the four candidate APLs were selected. In vitro experiments were performed to identify the relationship between APLs mInsA_ (2-10) DQ4 and mInsA_ (2-10) The relative affinity of HLA-A * 0201 molecules was relatively higher than that of the other two candidate APLs; these two APLs significantly inhibited mInsA (2-10) autoreactive CD8 + T cells compared to the native peptide group Proliferation. Two antagonistic APLs were administered to humanized NOD mice systematically. Only mInsA (2-10) DQ4 showed significant protective effect against T1D. Conclusion mInsA (2-10) DQ4 and mInsA 2- (2-10) DC6 are antagonistic APLs, whereas only mInsA 2- (2-10) DQ4 was found to have significant T1D protection in in vivo studies of humanized NOD mice Role, the next step will be to explore the immunological mechanism of mInsA_ (2-10) DQ4 prevention and treatment of T1D, which provides a good experimental basis for the clinical research of T1D prevention and control.