【摘要】 目的通过回输表达供体部分MHCⅡDRβ分子的自体骨髓细胞,观察受体对供体肝移植物的排斥情况。方法将供体猪MHCⅡDRβ分子进行克隆,并通过基因重组技术将其克隆到反转录病毒载体pLNCX2上,经过细胞学操作获得有感染性的病毒颗粒；体外感染受体骨髓细胞,并将感染后的骨髓细胞回输给受体,随后采用Nothern杂交和逆转录-聚合酶链反应(RT-PCR)等方法检测供体基因的表达,并通过MLR反应检测受体对供体的免疫排异反应；之后进行全肝脏移植实验,观测移植物的排异反应。结果实验组6头受体全部有外源基因的表达；进行回输后受体免疫功能都基本回复,MLR检测显示受体对供体的反应性明显下降,而对第三方的有较强的排异反应；进行移植后实验组中的5头移植物存活良好,而对照组于12～18周先后死亡,两者差异有统计学意义(P＜0．05)。结论猪受体在接受供体肝脏移植前,如果用供体的部分MHCⅡDRβ分子进行修饰并在体内进行表达,可以明显延长移植物的存活时间。更多还原

【Abstract】 Objective To investigate the tolerance of host pig to hepatitis graft from the donor whose MHCⅡDRβmolecule has been cloned and expressed by the host through back-infusion autologous bone marrow cells(BMC).Methods After MHCⅡDRβmolecules of donor pig were cloned,the genes were inserted into retroviral vector pLNCX2,and the infective virus particles were obtained by package cell AmphoPackTM-293.Following the BMC,which has been co-cultured with these infective virus parti- cles,back-infusion to recipient autologous,the expression of MHCⅡDrβgenes been showed by Northern blot and RT-PCR.Furthermore,the immune tolerance of the hosts to the donors has been detected by MLR(MLR,micro lymphocyte reaction).Results All six pigs received autologous BMC infected by virus and conditioned with growth factors and expressed MHCⅡDrβgenes of donor,and all pigs restitute its immunity system after some time of regiment.All pigs show very low ability reaction to donor’s lym- phocyte whose MHCⅡDrβbeen transduction.Comparing to the control group all pigs which expression donor’s MHCⅡDrβmolecμlars shows outstanding immune tolerance and life span.Conclusion Expres- sion of an allogeneic MHCⅡDrβthrough back-infusion BMC before hepatitis transplantation can elongate notably survive life of the graft.更多还原