四种嵌合免疫受体在T淋巴细胞中的表达分析

Analysis of expression for four chimeric immune receptors on human T lymphocyte

目的利用基因工程技术构建含人源抗癌胚抗原(CEA)单链抗体(scFv)的4种嵌合免疫受体(CIR),比较其在T淋巴细胞表面表达的情况。方法将构建好的嵌合免疫受体CIRZ、CIR1、CIR2、CIR3通过电穿孔方法分别转染人T淋巴细胞,流式细胞仪检测各嵌合免疫受体在T淋巴细胞的表达情况。结果有3种嵌合免疫受体成功表达在T细胞的表面,它们分别是CIR1、CIR2、CIR3,其中CIR3的表达更好。结论嵌合免疫受体能够在T淋巴细胞的表面有效表达,为下一步的体内实验打下基础,同时为这一治疗策略最终应用于临床提供可靠的实验依据。

Objective To compare the expressions of four chimeric immune receptors （CIRs） with anti-carcinoembryonic antigen （CEA）-scFv gene on human T lymphocyte via electroporation in vitro. Methods Four CIRs （CIRZ, CIR1, CIR2, CIR3） with anti-CEA- scFv gene were transfected into human peripheral blood mononuclear cells （PBMCs） obtained from healthy adults respectively. The expres- sions of surface molecules of CIRs were detected by flow eytometry after 16-hour transfection. Results Flow eytometry showed that CIRI, CIR2, CIR3 genes were efficiently expressed after transfection. But CIRZ cannot be detected any expression. Conclusions Three kinds of CIR genes can successfully express on human T lymphocytes surface. It can be the foundation of the further study and can provide the reliable evidence for clinical using this therapy.