RESEARCH ARTICLE

Novel T cells with improved in vivo anti-tumor activity generated by RNA electroporation

  • Xiaojun Liu 1,2 ,
  • Shuguang Jiang 2 ,
  • Chongyun Fang 2 ,
  • Hua Li 2 ,
  • Xuhua Zhang 2 ,
  • Fuqin Zhang 2 ,
  • Carl H. June 2,3 ,
  • Yangbing Zhao , 2,3
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  • 1. Key Laboratory for Experimental Teratology of Ministry of Education and Department of Immunology, School of Basic Medical Sciences, Shandong University, Jinan 250012, China
  • 2. Center for Cellular Immunotherapies, University of Pennsylvania Cancer Center, Philadelphia, PA 19104, USA
  • 3. Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104-5156, USA

Received date: 16 Feb 2017

Accepted date: 20 Apr 2017

Published date: 23 Aug 2017

Copyright

2017 The Author(s) 2017. This article is an open access publication

Abstract

The generation of T cells with maximal anti-tumor activities will significantly impact the field of T-cellbased adoptive immunotherapy. In this report, we found that OKT3/IL-2-stimulated T cells were phenotypically more heterogeneous, with enhanced anti-tumor activity in vivo 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.

Cite this article

Xiaojun Liu , Shuguang Jiang , Chongyun Fang , Hua Li , Xuhua Zhang , Fuqin Zhang , Carl H. June , Yangbing Zhao . Novel T cells with improved in vivo anti-tumor activity generated by RNA electroporation[J]. Protein & Cell, 2017 , 8(7) : 514 -526 . DOI: 10.1007/s13238-017-0422-6

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