Novel T cells with improved in vivo anti-tumor activity generated by RNA electroporation.
10.1007/s13238-017-0422-6
- Author:
Xiaojun LIU
1
;
Shuguang JIANG
2
;
Chongyun FANG
2
;
Hua LI
2
;
Xuhua ZHANG
2
;
Fuqin ZHANG
2
;
Carl H JUNE
2
;
Yangbing ZHAO
3
Author Information
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. Center for Cellular Immunotherapies, University of Pennsylvania Cancer Center, Philadelphia, PA, 19104, USA. Yangbing@upenn.edu.
- Publication Type:Journal Article
- Keywords:
CAR;
RNA electroporation;
T lymphocytes;
gene transfer;
manufacture
- MeSH:
Animals;
CD28 Antigens;
genetics;
immunology;
Electroporation;
Humans;
Immunity, Cellular;
Interleukin-2;
immunology;
K562 Cells;
Mice;
Muromonab-CD3;
immunology;
Neoplasms, Experimental;
genetics;
immunology;
pathology;
RNA, Messenger;
genetics;
immunology;
T-Lymphocytes;
immunology;
Tumor Necrosis Factor Receptor Superfamily, Member 9;
genetics;
immunology
- From:
Protein & Cell
2017;8(7):514-526
- CountryChina
- Language:English
-
Abstract:
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 co-electroporation 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.
