小鼠T淋巴细胞MHCI类抗原基因转染和表达的检测

A STUDY ON DETECTING GENE TRANSFECTION AND EXPRESSION OF MHC CLASS I GENE IN MICE T LYMPHOCYTES

摘要: 方法 :采用基因转移技术在体外将供体鼠MHCI类抗原H -2KbcDNA基因转染受体鼠T淋巴细胞 ,回输受体胸腺 ,诱导对供体的特异性耐受。对体外转染的受体鼠T淋巴细胞进行了基因整合和表达的检测。结果 :PCR扩增H -2KbcDNA ,证实目的基因已整合到染色体DNA上。取转染后 72h的T细胞 ,用抗H -2Kb 单克隆抗体荧光染色作流式细胞仪检测 ,证实在转染靶细胞膜上有H -2Kb 抗原表达 ,转染率为 46 2 %

Abstract: Methods: Donor mice MHC class I gene H-2K b cDNA was transferred into the spleen T lymphocytes of recipient strain by gene transfer technique and then the gene modified T cells returned to recipient mice thymus,to induce the donor-specific tolerance to allograft.In the present study,we have detected gene transfection and expression in T lymphocytes which transfered with donor gene H-2K b cDNA.DNA were prepared fron transfected T cells for PCR analysis.Result:H-2K b cDNA has integrated into chrosome DNA of T cells.After T cells were infected 72 hours they were analyzd on a flow cytometer using anti-mouse H-2K b monoclonal antibodies stain.The result showed that H-2K b gene has effectively expressioned and transfect efficiency was 46.2%.