Construction and functional analysis of EGFRvIII CAR-T cells co-expressing IL-15 and CCL19.
- Author:
Wanqiong CHEN
1
;
Na XIAN
2
;
Shaomei LIN
1
;
Wanting LIAO
1
;
Mingzhu CHEN
1
Author Information
- Publication Type:Journal Article
- Keywords: C-C chemokine ligand 19; chimeric antigen receptor T cell; glioblastoma; interleukin-15; the class III variant of the epidermal growth factor receptor
- MeSH: Humans; Receptors, Chimeric Antigen/metabolism*; Glioblastoma/metabolism*; Interleukin-15/metabolism*; Chemokine CCL19/metabolism*; Cell Line, Tumor; T-Lymphocytes/metabolism*
- From: Chinese Journal of Biotechnology 2023;39(9):3787-3799
- CountryChina
- Language:Chinese
- 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.
