关键词: T 细胞, 包膜蛋白, 融合白介素-2 的慢病毒载体, 转导效率, 过继免疫疗法

Abstract: Objective To construct the interleukin 2 ( IL2) -displaying lentivirus to improve the transduction efficiency of human primary T lymphocytes. Methods The IL2 gene was fused to the N-terminus of vesicular stomatitis virus G protein to produce the novel lentivirus-packaging plasmid pMD2.IL2-G. The yields of lentiviral particles pseudotyped with different doses of pMD2.IL2-G were compared using immunoblotting and ELISA. The infected capacity of IL2-displaying lentiviruses to T lymphocytes from healthy donors was tested using flow cytometry. Results The yield of lentiviral particles pseudotyped with pMD2.IL2-G was significantly less than that of wild-type lentiviruses packaged by pMD2.G. The mixture of pMD2.IL2-G and pMD2.G at a ratio of 1: 4 could obtain the equivalent yield of lentiviruses that of the wild-type, and significantly improve the transduction efficiency of enhanced green fluorescent protein into the T lymphocytes compared to the wild-type lentiviruses (89. 2 % vs. 40. 2 % ). Conclusion Our results demonstrate that IL2-displaying lentiviral particles have a greatly enhanced transduction efficiency into human T lymphocytes. This research is expected to provide strong support for the clinical application of genetically modified T cells.

Key words: T lymphocytes, envelope glycoprotein, interleukin-2-displaying lentiviruses, transduction efficiency, adoptive immunotherapy