标题：Novel T cells with improved in vivo anti-tumor activity generated by RNA electroporation
作者：Liu, Xiaojun; Jiang, Shuguang; Fang, Chongyun; Li, Hua; Zhang, Xuhua; Zhang, Fuqin; June, Carl H.; Zhao, Yangbing
作者机构：[Liu Xiaojun] Department of Immunology, School of Basic Medical Sciences, Shandong University, Key Laboratory for Experimental Teratology of Ministry 更多
通讯作者地址：[Zhao, YB]Univ Penn, Ctr Canc, Ctr Cellular Immunotherapies, Philadelphia, PA 19104 USA;[Zhao, YB]Univ Penn, Dept Pathol & Lab Med, Perelman Sch Med, 更多
关键词：T lymphocytes; CAR; manufacture; gene transfer; RNA electroporation
摘要：The generation of T cells with maximal anti-tumor activities will significantly impact the field of T-cell-based adoptive immunotherapy. In this report, we found that OKT3/IL-2-stimulated T cells were phenotypically more heterogeneous, with enhanced anti-tumor activity in vitro and when locally administered in a solid tumor mouse model. To further improve the OKT3/IL-2-based T cell manufacturing procedure, we developed a novel T cell stimulation and expansion method in which peripheral blood mononuclear cells were electroporated with mRNA encoding a chimeric membrane protein consisting of a single-chain variable fragment against CD3 and the intracellular domains of CD28 and 4-1BB (OKT3-28BB). The expanded T cells were phenotypically and functionally similar to T cells expanded by OKT3/IL-2. Moreover, co-electroporation of CD86 and 4-1BBL could further change the phenotype and enhance the in vivo anti-tumor activity. Although T cells expanded by the coelectroporation of OKT3-28BB with CD86 and 4-1BBL showed an increased central memory phenotype, the T cells still maintained tumor lytic activities as potent as those of OKT3/IL-2 or OKT3-28BB-stimulated T cells. In different tumor mouse models, T cells expanded by OKT3-28BB RNA electroporation showed anti-tumor activities superior to those of OKT3/IL-2 T cells. Hence, T cells with both a less differentiated phenotype and potent tumor killing ability can be generated by RNA electroporation, and this T cell manufacturing procedure can be further optimized by simply co-delivering other splices of RNA, thus providing a simple and cost-effective method for generating high-quality T cells for adoptive immunotherapy.