|Table of Contents|

Construction of T cells expressing chimeric antigen receptor targeting prostate stem cell antigen and its anti-tumor effect in tumor-bearing mice

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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

Issue:
2018年第12期
Page:
1053-1059
Research Field:
临床医学
Publishing date:

Info

Title:

Construction of T cells expressing chimeric antigen receptor targeting prostate stem cell antigen and its anti-tumor effect in tumor-bearing mice

Author(s):

HONG Juan CHEN Yunfan SHEN Junjie XU Yanmin QIAN Cheng

Biological Therapy Center, First Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400038, China

Keywords:

chimeric antigen receptor prostate stem cell antigen tumor immunotherapy lentivirus

PACS:
R322.64;R392-33;R73-362
DOI:
-
Abstract:

ObjectiveTo construct genetically modified T cells expressing chimeric antigen receptor (CAR) targeting prostate stem cell antigen (PSCA) and test the cytotoxicity of these cells against tumor cells both in vitro and in a tumorbearing mouse model. MethodsThe sequences encoding the light and heavy chain variable regions of a monoclonal antibody targeting PSCA were synthesized and integrated into lentiviral vectors using restriction enzymes. Human T lymphocytes isolated from peripheral blood mononuclear cells were infected with the recombinant lentivirus and the positivity rate of PSCACAR expression in the T lymphocytes was detected using flow cytometry. Cytotoxic assay was performed to evaluate the specific cytotoxicity of the genetically modified T cells against the PSCApositive HeLa cells and PSCAnegotive T24 cells, and ELISA was used to detect cytokine secretion by the cells. The antitumor effect of PSCACART cells was also tested in a NOG mouse model bearing HeLa cell xenograft. ResultsDNA sequencing analysis suggested correct synthesis of PSCACAR sequences, and enzyme digestion verified successful construction of the lentiviral vector PLPSCAG4H28TM28BBζ. The positivity rate of CAR expression was 53.9% in the T lymphocytes after infection with the lentiviral vector. The PSCACART cells showed a significantly stronger cytotoxicity against PSCApositive tumor cells than against PSCAnegative tumor cells (P<001) both in shortterm coincubation at a high effectivetarget ratio and in longterm coincubation at a low effectivetarget ratio. The PSCACART cells secreted significantly higher levels of cytokines than the control T cells upon stimulation by PSCApositive tumor cells (P<001), but the secretions were almost undetectable upon stimulation by PSCAnegative tumor cells. In the in vivo experiment, PSCACART cells exhibited a significant tumorsuppressive effect in the tumorbearing mice (P<001). ConclusionPSCACART cells capable of specific killing of PSCApositive tumor cells are successfully constructed both in vitro and in vivo.

References:


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Last Update: 2018-07-02