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:
Ye Di PU
1
;
Jia WANG
;
Qi DENG
;
Hai Bo ZHU
;
Yan Yu JIANG
;
Juan Xia MENG
;
Yu Ming LI
Author Information
1. Department of Hematology, Tianjin First Central Hospital, Tianjin 300192, China.
- Publication Type:Journal Article
- Keywords:
CAR-T Cells;
Expression of PD-1;
T cells;
Transfection efficiency;
mRNA level
- MeSH:
Antigens, CD19;
Humans;
Programmed Cell Death 1 Receptor/genetics*;
RNA, Messenger;
Receptors, Antigen, T-Cell;
T-Lymphocytes
- 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.
