关键词:基因转移;免疫耐受;皮肤移植;组织相容性抗原
【摘要 】 目的 通过胸腺内注射质粒PXN(N2-B19-H-2Kb ),表达外源性主要组织相容性抗原复合物(MHC)抗原,以小鼠皮肤移植为模型,诱导移植免疫耐受,为免疫耐受提供理论和实验依据。方法 应用逆转录病毒载体介导的基因转移技术,通过BALA/C胸腺内注射质粒PXN(N2-B19-H-2Kb ),将外源性MHC基因转移到胸腺细胞,14天后行异基因小鼠皮肤移植。应用聚合酶链反应(PCR),反转录聚合酶链反应(RT-PCR),单克隆抗体免疫荧光染色流式细胞仪检测胸腺细胞DNA,mRNA和MHC蛋白质表达。结果 外源性MHC基因已整合到靶细胞染色体DNA并有效地转录,胸腺细胞表面有外源性MHC分子表达,转染效率为5%,胸腺内注射质粒PXN(N2-B19-H-2Kb )能明显延长异基因小鼠的皮肤移植时间,平均27天,对照为10天(P<0.001)。受体鼠脾细胞对ConA的增殖反应在正常范围。结论 供体鼠MHC基因转移至受体鼠胸腺诱导了对供体皮肤移植的免疫耐受。
Expression of allogenic MHC gene inthymus by retroviral-mediated gene transfer induces tolerance
LI Jinhua,CHEN Yongxiong,YU Xueqing,etal.Nephrology Department,First Affiliated Hospital,Sun Yat-sen University of MedicalSciences,Guangzhou 510080
【Abstract 】 Objective Amice skin graft tolerence model was constructed by intrathymic injection of retroviralvector PXN(N2-B19-H-2Kb ) to express exogenous major histocompatibalitycomplex(MHC).It could add theoritical and experitmental data for tolerence.Methods Retroviralvector PXN(N2-B19-H-2Kb ) was injected into BALA/C thymus directly toexpress MHC of C57BL/6(H-2 b);and the polymerase chain reaction (PCR) , the reversetranscription polymerase chain reaction (RT-PCR) and flow cytometry assays were used tocheck DNA,mRNA and MHC protein expression in thymocytes.Results AllogenicMHC gene has been integrated into the chromosomal DNA of BALA/C thymus and expressedstably ;it induced allogenic mouse skin graft tolerance:27days vs 10 days(P<0.001).Thereaction of splenocytes in tolerance mice to ConA was in normal range.Conclusion AllogenicMHC gene transferring into recepient thymus directly could induce tolerance to allogenicmice.
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