Effects of sodium iodide symporter co-expression on proliferation and cytotoxic activity of chimeric antigen receptor T cells in vitro.
10.12122/j.issn.1673-4254.2022.07.15
- Author:
Chuanhuizi TIAN
1
;
Pu Feng HUANG
1
;
Yu Jia HE
1
;
Liang WANG
1
;
Zhi Ping PENG
1
Author Information
1. Department of Radiation Medicine, College of Basic Medicine, Chongqing Medical University, Chongqing 400014, China.
- Publication Type:Journal Article
- Keywords:
chimeric antigen receptor T cells;
co-expression;
killing activity;
proliferation in vitro;
sodium iodide symporter
- MeSH:
Antineoplastic Agents;
Cell Line, Tumor;
Cell Proliferation;
Iodine;
Lymphotoxin-alpha;
Receptors, Chimeric Antigen;
Symporters;
T-Lymphocytes
- From:
Journal of Southern Medical University
2022;42(7):1062-1068
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effects of co-expression of sodium iodide symporter (NIS) reporter gene on the proliferation and cytotoxic activity of chimeric antigen receptor (CAR)-T cells in vitro.
METHODS:T cells expressing CD19 CAR (CAR-T cells), NIS reporter gene (NIS-T cells), and both (NIS-CAR-T cells) were prepared by lentiviral infection. The transfection rates of NIS and CAR were determined by flow cytometry, and the cell proliferation rate was assessed using CCK-8 assay at 24, 48 and 72 h of routine cell culture. The T cells were co-cultured with Nalm6 tumor cells at the effector-target ratios of 1∶2, 1∶1, 2∶1 and 4∶1 for 24, 48 and 72 h, and the cytotoxicity of CAR-T cells to the tumor cells was evaluated using lactate dehydrogenase (LDH) assay. ELISA was used to detect the release of IFN-γ and TNF-β in the co-culture supernatant, and the function of NIS was detected with iodine uptake test.
RESULTS:The CAR transfection rate was 91.91% in CAR-T cells and 99.41% in NIS-CAR-T cells; the NIS transfection rate was 47.83% in NIS-T cells and 50.24% in NIS- CAR-T cells. No significant difference in the proliferation rate was observed between CAR-T and NIS-CAR-T cells cultured for 24, 48 or 72 h (P> 0.05). In the co-cultures with different effector-target ratios, the tumor cell killing rate was significantly higher in CAR-T group than in NIS-CAR-T group at 24 h (P < 0.05), but no significant difference was observed between the two groups at 48 h or 72 h (P>0.05). Higher IFN-γ and TNF-β release levels were detected in both CAR-T and NIS-CAR-T groups than in the control group (P < 0.05). NIS-T cells and NIS-CAR-T cells showed similar capacity of specific iodine uptake (P>0.05), which was significantly higher than that in the control T cells (P < 0.05).
CONCLUSION:The co-expression of the NIS reporter gene does not affect CAR expression, proliferation or tumor cell-killing ability of CAR-T cells.
