Functional investigation of chimeric antigen receptor T cells targeting LMP1 antigen

• Published in: Zhōnghuá xuèyèxué zázhì 2022-03, Vol.43 (3), p.229-234
• Main Authors: He, H Z, Xing, Y Y, Zhang, Y, Xu, Y X, Tian, Z, Xing, H Y, Tang, K J, Rao, Q, Wang, J X, Wang, M
• Format: Article
• Language: Chinese
• Subjects: Cell Line, Tumor; Herpesvirus 4, Human; Humans; Lentivirus; Lymphoma - therapy; Receptors, Chimeric Antigen - genetics; T-Lymphocytes; Viral Matrix Proteins
• ISSN: 0253-2727
• DOI: 10.3760/cma.j.issn.0253-2727.2022.03.008

This study aimed to create a type of CAR-T cells that targets LMP1 antigen and study its immunotherapeutic effect on LMP1-positive hematological malignancies. To generate LMP1 CAR-T cells, a plasmid expressing LMP1 CAR was created using molecular cloning technology, and T cells were infected with LMP1 CAR lentivirus. The effects of LMP1 CAR-T cells on specific cytotoxicity against LMP1-positive tumor cell lines infected with the EB virus had been confirmed. ① LMP1 protein expressing on EB virus-positive lymphoma cells surface was verified. ② The LMP1 CAR-expressing plasmid was created, and LMP1 CAR-T cells were obtained by infecting T cells with a lentivirus packaging system, with an infection efficiency of more than 80% . ③LMP1 CAR-T cells have a 4∶1 effect-to-target ratio in killing LMP1-positive lymphoma cells. The killing effect of LMP1 CAR-T cells on Raji cells was enhanced after 48 h of coculture, but there was no significant killing effect on Ramos, which are LMP1-negative lymphoma cells. ④After cocult