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Generation, ex vivo expansion and safety of engineered PD1-knockout primary T cells from cynomolgus macaques.

Abstract：

:Programmed cell death protein 1 (PD1) is a cell-surface receptor that plays a vital regulatory role in suppressing inflammatory T cell activity; therefore, it is an ideal target for T cell-redirecting therapies. Here, we describe a cynomolgus macaque model for studying the transfer of PD1-modified T cells. We developed the first T cell engager targeting the disruption of PD1 by electroporation of plasmids encoding sgRNA and Cas9. There were no significant differences between mock T cells and PD1-knockout (PD1-KO) T cells in terms of cell viability, T cell signature marker expression, cell apoptosis, or cell cycling during prolonged in vitro culture. However, in a mixed lymphocyte reaction, PD1-KO T cells exhibited increased proliferation for both CD4+ and CD8+T cells and enhanced IFNγ release. We adoptively transferred autologous PD1-KO T cells into three cynomolgus monkeys. The PD1-KO T cells did not cause overt toxicity as measured by evaluating body weight, hematological parameters, and blood chemistry parameters. Histopathological analyses of tissues showed no lesions related to the infused PD1-KO T cells. Our findings demonstrate the utility of cynomolgus monkeys in expanding PD1-KO T cells and evaluating the safety of this immunotherapy and provide a new strategy for T cell-based adoptive cell therapies.

journal_name

Mol Immunol

journal_title

Molecular immunology

authors

Gao CE,Song Q,Zhang M,Li J,Miao Y,Li Z,Dong J

doi

10.1016/j.molimm.2020.05.007

subject

Has Abstract

pub_date

2020-08-01 00:00:00

pages

100-108

eissn

0161-5890

issn

1872-9142

pii

S0161-5890(20)30348-5

journal_volume

124

pub_type

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