PD-1 expression, mRNA level and cytotoxicity changes in CD19CAR-T cells
10.3760/cma.j.issn.0253-2727.2019.09.010
- VernacularTitle: CD19 CAR-T细胞培养过程中PD-1蛋白、mRNA水平及细胞杀伤活性变化
- Author:
Yedi PU
1
;
Jia WANG
;
Qi DENG
;
Haibo ZHU
;
Yanyu JIANG
;
Juanxia MENG
;
Yuming LI
Author Information
1. Department of Hematology, Tianjin First Central Hospital, Tianjin 300192, China
- Publication Type:Journal Article
- Keywords:
Expression of PD-1;
mRNA level;
Transfection efficiency;
T cells;
CAR-T Cells
- From:
Chinese Journal of Hematology
2019;40(9):759-763
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To observe the changes of PD-1 expression, mRNA level and cytotoxic activity of CD19 CAR-T cells during the culture process of CAR-T cells.
Methods:The peripheral blood T cells of 6 lymphoma patients with high expression of PD-1 and 6 healthy volunteers were the source of CAR-T cells. The expression of PD-1 was analyzed by flow cytometry. The mRNA level of PD-1 was analyzed by PCR. The cell proliferation was analyzed by CCK-8 assay. The cytotoxicity was analyzed by LDH assay.
Results:①The transfection efficiency of high PD-1 expression T cells and healthy volunteer T cells were as the same (P>0.05) . ②The cell proliferation capacity of CD19 CAR-T cells from high PD-1 expression T cells or healthy volunteer T cells, with or without PD-1 inhibitor were as the same (P>0.05) . ③The cytotoxicity to lymphoma cells of high PD-1 expression T cells and CAR-T cells were lower than that of these two T cells combined with PD-1 inhibitor and the CAR-T cells from healthy volunteer T cells (P<0.001) . There was no difference of the cytotoxicity between the CAR-T cells from high PD-1 expression T cells combined with PD-1 inhibitor and the CAR-T cells from healthy volunteer (P>0.05) . ④There was no difference of the expression of PD-1 in all CAR-T cell groups during the culture process (P>0.05) . There was no difference of mRNA level of PD-1 in all groups during the culture process (P>0.05) . ⑤The PD-1 expression of CAR-T cells increased by the time of culture after contacting with lymphoma cells (P<0.001) . The PD-1 inhibitors could antagonize this effect. There was no difference of mRNA level of PD-1 in all groups after contacting with lymphoma cells (P>0.05) .
Conclusion:The PD-1 expression of CAR-T cells from high PD-1 expression T cells increased by the time of culture after contacting with lymphoma cells. However, the mRNA level of PD-1 of all groups did not change, even if PD-1 inhibitor was applied.
