Construction and functional analysis of EGFRvIII CAR-T cells co-expressing IL-15 and CCL19 / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 3787-3799, 2023.
Article
in Chinese
| WPRIM
| ID: wpr-1007993
ABSTRACT
The aim of this study was to investigate the functional characteristics and in vitro specific killing effect of EGFRvIII CAR-T cells co-expressing interleukin-15 and chemokine CCL19, in order to optimize the multiple functions of CAR-T cells and improve the therapeutic effect of CAR-T cells targeting EGFRvIII on glioblastoma (GBM). The recombinant lentivirus plasmid was obtained by genetic engineering, transfected into 293T cells to obtain lentivirus and infected T cells to obtain the fourth generation CAR-T cells targeting EGFRvIII (EGFRvIII-IL-15-CCL19 CAR-T). The expression rate of CAR molecules, proliferation, chemotactic ability, in vitro specific killing ability and anti-apoptotic ability of the fourth and second generation CAR-T cells (EGFRvIII CAR-T) were detected by flow cytometry, cell counter, chemotaxis chamber and apoptosis kit. The results showed that compared with EGFRvIII CAR-T cells, EGFRvIII-IL-15-CCL19 CAR-T cells successfully secreted IL-15 and CCL19, and had stronger proliferation, chemotactic ability and anti-apoptosis ability in vitro (all P < 0.05), while there was no significant difference in killing ability in vitro. Therefore, CAR-T cells targeting EGFRvIII and secreting IL-15 and CCL19 are expected to improve the therapeutic effect of glioblastoma and provide an experimental basis for clinical trials.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
T-Lymphocytes
/
Glioblastoma
/
Interleukin-15
/
Cell Line, Tumor
/
Chemokine CCL19
/
Receptors, Chimeric Antigen
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2023
Type:
Article
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