融合IL2的慢病毒实现对T细胞的高效基因转导

Potent Gene Transfer in Human Primary T Lymphocytes Using Interleukin-2-displaying Lentiviral Vectors

目的制备融合IL2的慢病毒以提高对人原代T淋巴细胞的转导效率。方法将白细胞介素-2(IL2)基因定向克隆至水疱性口炎病毒G蛋白(VSV-G)编码基因5′端,构建新型慢病毒包装辅助质粒pMD2.IL2-G。使用不同剂量pMD2.IL2-G进行慢病毒包装,应用免疫印迹和ELISA比较病毒滴度,流式细胞术检测病毒对T细胞的转导效率。结果单独使用pMD2.IL2-G显著降低病毒得率。与传统慢病毒相比,pMD2.IL2-G与原始辅助质粒pMD2.G以1∶4混合制备的慢病毒,能够显著提高EGFP基因导入T细胞的效率(89.2%vs.40.2%),且病毒得率相当。结论融合IL2的慢病毒能够对T细胞进行高效的基因转导,为基因修饰T细胞的临床应用提供有力支持。

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 plas-mid 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 lentivi-ral 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 effi-ciency of enhanced green fluorescent protein into the T lymphocytes compared to the wild-type lenti-viruses(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.