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MHC基因胸腺转移诱导异基因小鼠皮肤移植免疫耐受

Expression of allogenic MHC gene in thymus by retroviralmediated gene transfer induces tolerance

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【作者】 李锦华陈永雄余学清张仕光

【Author】 LI Jinhua,CHEN Yongxiong,YU Xueqing,et al.Nephrology Department,First Affiliated Hospital,Sun Yatsen University of Medical Sciences,Guangzhou 510080

【机构】 中山医科大学附属第一医院肾内科

【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。

【Abstract】 Objective A mice skin graft tolerence model was constructed by intrathymic injection of retroviral vector PXN(N2B19H2Kb) to express exogenous major histocompatibality complex(MHC).It could add theoritical and experitmental data for tolerence . Methods Retroviral vector PXN(N2B19H2Kb) was injected into BALA/C thymus directly to express MHC of C57BL/6(H2 b);and the polymerase chain reaction (PCR) , the reverse transcription polymerase chain reaction (RTPCR) and flow cytometry assays were used to check DNA,mRNA and MHC protein expression in thymocytes. Results Allogenic MHC gene has been integrated into the chromosomal DNA of BALA/C thymus and expressed stably ;it induced allogenic mouse skin graft tolerance:27days vs 10 days(P<0001).The reaction of splenocytes in tolerance mice to ConA was in normal range. Conclusion Allogenic MHC gene transferring into recepient thymus directly could induce tolerance to allogenic mice.

【基金】 国家自然科学基金
  • 【文献出处】 中华肾脏病杂志 ,CHINESE JOURNAL OF NEPHROLOGY , 编辑部邮箱 ,1999年03期
  • 【分类号】R622
  • 【被引频次】3
  • 【下载频次】78
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