间接抗原递呈途径在表皮细胞免疫原性中作用的实验研究

An experimental study on the role of indirect antigen presentation pathway on the immunogenicity of epidermal cells

目的 探讨间接抗原递呈途径在体外培养的同种异体表皮细胞免疫原性中的作用。 方法 体外分离、培养人表皮细胞 (HEC)、异体人外周血淋巴细胞 (PBL)和单个核细胞 (PBM ,含单核细胞 )。采用人源毒性T淋巴细胞相关抗原 4(CTLA4Ig)腺病毒表达载体转染HEC ,观察CTLA4Ig的表达情况。未转染及已转染的HEC中 ,均加入异体PBL或PBM共培养 ,并以单独培养的PBL或PBM为对照。应用氚标记胸腺嘧啶脱氧核苷 ( 3H TdR)掺入法 ,测定各培养体系中PBL、PBM增殖情况。 结果 CTLA4Ig腺病毒表达载体能成功转染HEC并使其表达相应蛋白。未转染的HEC能刺激异体PBL轻度增殖 (P <0 .0 5 ) ,以及含单核细胞的异体PBM明显增殖 (P <0 .0 1) ;经CTLA4Ig基因修饰的HEC刺激PBL、PBM后 ,两者的增殖反应明显减弱 (P <0 .0 5 )。 结论 间接抗原递呈途径在HEC免疫原性中发挥了重要作用 ,CTLA4Ig能明显抑制该作用。

Objective To explore the role of indirect antigen presentation pathway on the immunogenecity of epidermal cells. Methods Human epidermal cells (HEC), allogeneic human peripheral blood lymphocytes (PBL) and mononuclear cells (PBM, including monocytes) were isolated and cultured in vitro. HECs were transfected by human-originated CTLA4Ig-adenovirus vector. The CTLA4Ig expression was observed. Allogeneic PBLs or PBMs were added to the transfected and non-transfected HECs with simple cultured PBLs and PBMs as the control. The proliferation of PBL and PBM was determined by 3 H-TdR incooperation. Results HECs could be successfully transfected by CTLA4Ig-adenovirus vector and expressed corresponding proteins. The non-transfected HECs could stimulate slight proliferation of allogeneic PBLs (P<0.05) and stimulate remarkable proliferation of PBMs (including monocytes) (P<0.05). The proliferation reaction of PBLs and PBMs decreased significantly (P<0.05) after being stimulated by HEC which was modulated by CTLA4Ig genes. Conclusion Indirect antigen presentation pathway might play important roles in the HEC immunogenicity which could be evidently inhibited by CTLA4Ig. [