供心体外转染腺病毒介导的CD40Ig融合基因延长小鼠移植心脏存活时间  

Donor heart gene transfer of CD40Ig mediated by adenovirus to prolong the survival time of murine cardiac allograft

作　　者：黄保军[1] 尹辉[1] 黄亚非[1] 徐军发[2] 熊平[1] 冯玮[1] 郑芳[1] 徐勇[1] 方敏[1] 龚非力[1] 

机构地区：[1]华中科技大学同济医学院免疫学系 [2]广东医学院临床免疫学教研室

出　　处：《中华器官移植杂志》2006年第9期524-527,共4页Chinese Journal of Organ Transplantation

基　　金：国家重点基础研究发展规划(973)资助项目(No.2001cb500008)

摘　　要：目的探讨体外转染CD40Ig融合基因对小鼠移植心脏存活时间的影响。方法构建携带小鼠CD40胞外段和人IgGFc融合基因的重组腺病毒载体(AdCD40Ig),以BALB/c小鼠为供者,C57BL/6小鼠为受者,建立小鼠腹部异位心脏移植模型,实验组供心移植前在体外以AdCD40Ig灌注,转染CD40Ig基因,另设空载体转染对照组、非转染对照组和近交系对照组(供、受者均为近交系C57BL/6小鼠)。术后观察移植心的存活及移植物中炎症细胞浸润情况,采用酶联免疫吸附试验(ELISA)检测受者体内CD40Ig融合蛋白表达情况,流式细胞仪检测受者体内产生γ干扰素(IFN-γ)的脾细胞。结果实验组移植心的存活时间达(15.8±0.7)d,明显长于空载体转染对照组和非转染对照组(P<0.01)。术后第2d,实验组受者体内CD40Ig融合蛋白表达最高,1周后明显降低。术后第7d,实验组移植心组织中浸润的炎症细胞明显比未处理对照组和空载体对照组少。实验组产生IFN-γ的CD4+和CD8+T淋巴细胞分别为(2.18±0.16)%和(10.82±0.74)%,与近交系对照组接近,明显低于未处理对照组和空载体对照组(P<0.01)。结论供心体外转染CD40Ig融合基因可有效抑制移植后受者体内同种T淋巴细胞的增殖,并延长移植心的存活时间。Objective To explore the effects of CD40Ig gene transfer in vitro on cardiac allograft survival in mice. Methods The recombinant adenovirus vector carrying murine CD40 extracellular domain and human IgG Fc fusion gene （AdCD40Ig） was constructed. Using BAI.B/c mice as donors and C57BL/6 mice as recipients, the model of mice abdomen heterotopical heart transplantation was set up. In the experimental group, the isolated donor heart was transfected with AdCD40Ig, then transplanted to recipient. The other groups included empty vector transfected control group, untransfected control group, and syngeneic control group （the donors and the recipients were C57BL/6 mice）. The survival time of donor hearts and the infiltration of inflammatory cells in heart allograft were observed. The expression of CD40lg fusion protein in recipient was identified by Sandwich ELISA; The IFN-γ producing cells in recipient was determined by FACS. Results The survival time of the cardiac allograft in experimental group was （15.8 ± 0.7） days, significantly longer than empty vector transfected control group and untransfected control group （P〈0. （11）. In the second day after transplantation, ELISA confirmed that the expression of CD40Ig fusion protein in recipients was highest, but decreased rapidly after one week; On the 7th day after transplantation, the infiltration of inflammatory cells of allograft in experimental group was less than empty vector transfected control group and untransfected control group. The IFN-γ producing CD4^＋ and CD8 ^＋ T cells in experimental group were （2. 18 ±0. 16） % and （10. 82 ± 0. 74）%, respectively, which was close to syngeneic control group, but signi-ficantly lower than empty vector transfected control group and untransfected control group （P〈0. 01）. Conclusion Donor heart transfected with AdCD40Ig in vitro could sufficiently inhibit the proliferation of alloreactive T cells in recipients and prolong the murine cardiac allograft survival.

关 键 词：心脏移植 CD40配体 腺病毒科 转染 

分 类 号：R654.2[医药卫生—外科学]