结核分枝杆菌来源HSP65引发复发性口腔炎的表位通过细胞免疫加剧动脉粥样硬化  

Cellular Immunity to an Epitope from Recurrent Aphthous Stomatitis Enhances Atherosclerosis in rRabbit and LDL Receptor-Deficient Mice

作　　者：张宇[1] 熊祺琰[1] 胡向兵[1] 孙云霄[1] 张会勇[1] 曹荣月[1] 刘景晶[1] 

机构地区：[1]中国药科大学生命科学与技术学院,江苏南京210009

出　　处：《药物生物技术》2010年第4期291-296,共6页Pharmaceutical Biotechnology

基　　金：国家自然科学基金(30872393;30672464;30701023)

摘　　要：研究结核分枝杆菌来源的热休克蛋白65(mHSP65)中复发性口腔炎(RAS)和白塞病(BD)表位在动脉粥样硬化中的作用及免疫系统反应类型。化学合成各个表位肽段,在高血胆固醇兔子模型中筛选出加剧动脉粥样硬化的表位为RAS相关表位mHSP6591-105;用抗原亲和柱纯化特异性抗体,western-blot表明抗体与mHSP65及人热休克蛋白60都能结合;激光共聚焦研究特异性抗体在HUVECs中的结合部位分布。发现该表位出现在胞浆和凋亡的HUVECs表面,且亲和纯化获得的抗体不能诱导热休克后HUVECs的凋亡及细胞周期变化;在ldlr-/-小鼠模型中发现mHSP6591-105反应性脾细胞IFN-γ分泌显著增加,过继性转移脾细胞加速了动脉粥样硬化过程。复发性口腔炎表位引起的细胞免疫反应在加速动脉粥样硬化过程中起到了重要作用。Autoimmune diseases such as atherosclerosis, recurrent aphthous stomatitis （RAS） and Behcet＇s disease （BD） are the common diseases worldwide. Furthermore, RAS and BD both have overlapped epitopes with atherosclerosis in mHSP65. However, the roles which RAS and BD play on the progress of atherosclerosis are not clear. Our research might make pepople better understand the susceptible population of atherosclerosis. High fat and high cholesterol food fed rabbits, wide-type mice and ldlr-/- mice were used. Firstly, high fat and high cholesterol food fed rabbits were used to investigate the atherogenic role of four epitopes from mHSP65, and the western-blot determined that specific antibodies cross-react with recombinant human heat shock protein-60. Secondly, scanning electron microscope showed that immune response induced by mHSP6591-105 has direct toxic effect on endothelium of wide-type mice. However antibodies to mHSP65 91-105 can not induce apoptosis nor change the cell cycle of HUVECs. Cellular distribution of special epitopes in HUVECs was observed by laser scanning confocal microscopy. Thirdly, the adoptive transfer of mHSP65 91-105 specific splenic cells can accelerate the atherosclerosis in ldlr-/- model, showing that specific T cells play an important role. Also the significantly raised level of IFN - y secreted by these T cells was detected. Administration of mHSP65 91-105 accelerates atherosclerosis in animal models and the Th1 response plays the important role in the progress. The results linked RAS with ath erosclerosis, and it might be valuable to define susceptible population of atherosclerosis.

关 键 词：动脉粥样硬化 热休克蛋白 特异性抗体 细胞免疫 过继性转移 

分 类 号：R965[医药卫生—药理学]