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目的将外源人免疫球蛋白IgGFc段Ⅱ型受体CD32a分子表达于K562细胞表面,为构建人工抗原递呈细胞提供重要前提。方法自U937细胞RT-PCR得到CD32a的cDNA基因,与T载体连接后亚克隆入表达载体pcDNA3.1(+)。经测序后利用脂质体介导的转染将CD32a分子表达在K562细胞的表面。结果构建的T载体测序后,Genebank比对证实为CD32a的cDNA分子,免疫荧光和流式细胞仪结果均说明CD32a分子在K562细胞得到表达(96.9%)。结论获得的人工构建的表达人免疫球蛋白IgGFc受体的K562细胞,完成人工抗原递呈细胞制备的首要步骤。
Objective To express CD32a, an exogenous immunoglobulin IgGFc segment Ⅱ receptor, on the surface of K562 cells and provide an important precondition for the construction of artificial antigen-presenting cells. Methods cDNA of CD32a was obtained by RT-PCR from U937 cells and subcloned into the expression vector pcDNA3.1 (+) after it was ligated with T vector. After sequencing, the CD32a molecule was expressed on the surface of K562 cells by liposome-mediated transfection. Results After the constructed T vector was sequenced, the cDNA of CD32a was confirmed by Genebank comparison. The results of immunofluorescence and flow cytometry showed that CD32a was expressed in K562 cells (96.9%). CONCLUSIONS: The constructed artificial K562 cells expressing human immunoglobulin IgGFc receptor are the first steps in the preparation of artificial antigen-presenting cells.