【作者】 于倩；

【导师】 吴雄文；

【作者基本信息】 华中科技大学 ， 免疫学， 2009， 硕士

【摘要】 同种异型移植排斥反应所识别的靶抗原主要是表达于移植物细胞表面的主要组织相容性复合物(MHc)分子，由于几类主要组织相容性复合物(HLA)基因的丰富多态性，很难在几体水平研究单个异体HLA分子在移植排斥中的具体作用。HLA．A2转基因小鼠的出现为体内研究移植排斥反应机制与干预措施提供了理想的动物模型。本课题以c57BL／6来源的HLA-A2转基因小鼠(以下简称A2小鼠)为主要实验对象，建立仅HLA-A2基因差别所引起的皮肤移植排斥反应动物模型，并通过比较HLA-A2基因差别所引起的移植排斥反应与典型急性排斥反应和慢性排斥反应的异同，探讨这种移植排斥反应的类型与机制。本研究的主要内容及结果如下：1利用HLA-A2特异性引物RT-PcR及HLA-A2分子特异性抗体BB7.2检测显示HLA-A2转基因小鼠细胞表面表达HLA-A2分子。2以普通c57BL/6雄性小鼠为受者，雄性A2小鼠为供者，建立仅HLA-A2基因差别所引起的皮肤移植排斥反应动物模型，结果发现针对A2小鼠皮肤移植物的排斥反应发生迅速，9～13天出现皮片排斥反应。将两者脾细胞进行混合淋巴细胞培养结果显示c57BL／6来源的T细胞在体外受A2小鼠的细胞刺激后明显增殖。3与同种异基因小鼠皮肤移植所引起的急性排斥反应及雌雄异体移植所引起的慢性排斥反应相比较，针对HLA-A2转基因小鼠皮肤移植物的排斥反应及体外同种反应表现出发生迅速，反应强度大等急性排斥反应的特点。由于供受者之间仅存在HLA-A2的差别，而针对A2小鼠皮肤移植排斥反应更接近急性排斥反应，因此该动物模型可用于进一步认识移植排斥中直接识别机制和探讨移植排斥反应的防治手段。更多还原

【Abstract】 The antigens detected by T cells in allograft rejection are mostly derived from a highly polymorphic region of the genome called the major histocompatibiloity complexe (MHC). The concrete efforts of single MHC molecule in allograft rejection are not easy to understant thoroughly for the MHC polymorphism. The HLA-A2 transgenic mouse provides us with a possible model to understand the graft rejection. We have used the HLA-A2 transgenic mouse of C57BL/6 origin (A2 mouse) as donors to test the rejection of skin grafts, in order to find a in vivo model of rejection against HLA-A2, therefore, the model can be used to study the mechanisms of acute rejection and to develop new approaches to treat and prevent the rejection. In this study, we establish the skin graft model from A2 mouse to normal C57BL/6 mouse though back to back full-thickness skins transplantation. The rejection was observed and compared with the the acute rejection of allogeneic skins transplantation and the chronic one across the sex. The results list below:1 -, The expression of HLA-A2 molecule on the transgenic mouse cell’s surface was detected by both RT-PCR with specific primers of HLA-A2 extra-cellular domain and the HLA-A2 allelic specific antibody BB7.2.2^ Set up the skin graft animal model (recipient: normal C57BL/6 male mice, donor: A2 male mice) in which HLA-A2 molecule is expected to be the sole rejection antigen. Results showed that A2 mouse skin grafts were rapidly rejected in 9—13 days, and the T cells proliferation of the recipient was stronger in mixed lymphocyte culture.3 -, Compared to the chronic rejection reaction caused by the unmatch sex between recipient and donor, the reaction against the HLA-A2 molecule is more likely to be the acute rejection of allogeneic skin graft, because which occurs rapid in time and intensity in inflammatory response.The rejection against HLA-A2 molecule in the animal model provides us with an in vivo model which would be useful for us to further understand the direct and the indirect recognition in the graft rejection and to induce the graft tolerance.更多还原