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目的 建立一个在胸腺内表达外源性主要组织相容性抗原复合体(MHC)抗原的实验模型,为下一步研究作准备。方法 应用逆转录病毒载体介导的基因转移技术,首次将外源性MHC基因转移到T淋巴表达并应用聚合酶链反应(PCR)及反转录聚合酶链反应(RT-PCR)检测转染T细胞DNA及mRNA。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;单克隆抗体免疫荧光染色流式细胞仪检测显示在转染T细胞膜有外源性MHC分子表达;转染效率为46.2％。结论 外源性MHC基因可以转移到T细胞稳定表达,为今后的研究提供了经验。
Objective To establish an experimental model for expression of exogenous major histocompatibility complex (MHC) antigen in the thymus, and to prepare for further study. Methods Using retroviral vector-mediated gene transfer technology, the exogenous MHC gene was first transferred to T lymphocytes and transfected by RT-PCR and polymerase chain reaction (RT-PCR) T cell DNA and mRNA. Results The exogenous MHC gene was integrated into the chromosomal DNA of target cells and transcribed efficiently. The expression of exogenous MHC molecules was detected by immunofluorescence staining and flow cytometry. The transfection efficiency was 46.2%. Conclusion Exogenous MHC gene can be transferred to T cells for stable expression, which provides experience for future research.