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目的:通过胸腺内注射质粒 PXN(N2—B19—H—2K~b),表达外源性主要组织相容性抗原复合物(major histo-compatibility complex,MHC)抗原,为下一步诱导异基因小鼠器官移植耐受作准备.方法:通过 BALB/C 小鼠胸腺内注射质粒 PXN(N2—B19—H—2K~b),将外源性的编码 C57BL/6小鼠MHCI类抗原的H—2K~bcDNA转移到 BALB/C小鼠胸腺,用聚合酶链反应(PCR),反转录聚合酶链反应(RT—PCR),单克隆抗体免疫萤光染色流式细胞仪检测BALB/C小鼠胸腺细胞DNA,mRNA和MHC蛋白质表达,结果:BALB/C小鼠胸腺表面有外源性MHC分子表达,转染效率为5.1%结论:通过胸腺内注射逆转录病毒载体PXN(N2—B19—H—2K~b),小鼠胸腺能表达同种MHC基因,为下一步行异基因小鼠器官移植耐受打下了物质基础.
OBJECTIVE: To express the major histocompatibility complex antigen (MHC) by intra-thymus injection of plasmid PXN (N2-B19-H-2K ~ b) Mouse organ transplantation tolerance.Methods: The exogenous DNA encoding the MHC class I antigens of C57BL / 6 mice was injected into the thymus of BALB / C mice via the plasmid PXN (N2-B19-H-2K ~ b) 2K ~ bcDNA was transferred to the thymus of BALB / C mice. The BALB / C mice were detected by polymerase chain reaction (PCR), reverse transcription polymerase chain reaction (RT-PCR) and monoclonal antibody immunofluorescence staining The results showed that exogenous MHC was expressed on the surface of thymus of BALB / c mice and the transfection efficiency was 5.1%. CONCLUSION: The retroviral vector PXN (N2-B19- H-2K ~ b), mouse thymus can express the same kind of MHC gene, laid the material foundation for the tolerance of organ transplant in the next walking allogeneic mice.