Preparation of CD52-targeted chimeric antigen receptor-modified T cells and their anti-leukemia effects

• Published in: Zhōnghuá xuèyèxué zázhì 2022-04, Vol.43 (4), p.279-286
• Main Authors: Liu, Y, Tang, K J, Chen, Z Q, Mou, J L, Xu, Y X, Xing, H Y, Tian, Z, Rao, Q, Wang, M, Wang, J X
• Format: Article
• Language: Chinese
• Subjects: CD52 Antigen; Cell Line, Tumor; Humans; Immunotherapy, Adoptive - methods; Lentivirus - genetics; Leukemia; Receptors, Antigen, T-Cell; Receptors, Chimeric Antigen - genetics
• ISSN: 0253-2727
• DOI: 10.3760/cma.j.issn.0253-2727.2022.04.003

To construct chimeric antigen receptor (CAR) T cells targeting CD52 (CD52 CAR-T) and validate the effect of CD52 CAR-T cells on CD52-positive leukemia. A second-generation CD52-targeting CAR bearing 4-1BB costimulatory domain was ligated into a lentiviral vector through molecular cloning. Lentivirus was prepared and packaged by 293 T cells with a four-plasmid system. Fluorescein was used to label cell surface antigens to evaluate the phenotype of CD52 CAR-T cells after infection. Flow cytometry and ELISA were used to evaluate the specific cytotoxicity of CD52 CAR-T cells to CD52-positive cell lines in vitro. ①A pCDH-CD52scFv-CD8α-4-1BB-CD3ζ-GFP expressing plasmid was successfully constructed and used to transduce T cells expressing a novel CD52-targeting CAR. ②On day 6, CD52-positive T cells were almost killed by CD52-targeted CAR-T post lentivirus transduction [CD52 CAR-T (4.48 ± 4.99) %, Vector-T (56.58±19.8) %, =0.011]. ③T cells transduced with the CAR targeting CD52 showed low levels of apoptosis and coul