An experimental study of CD4 targeted chimeric antigen receptor modified T cell with anti-lymphoma activity
10.3760/cma.j.issn.0253-2727.2018.02.014
- VernacularTitle: 针对CD4膜蛋白的CAR-T细胞抗T细胞淋巴瘤的活性研究
- Author:
Guanghua CHEN
1
;
Haiwen HUANG
;
Yi WANG
;
Huiwen LIU
;
Liangjing XU
;
Xiao MA
;
Shengli XUE
;
Xuefeng HE
;
Ying WANG
;
Bin GU
;
Caixia LI
;
Huiying QIU
;
Xiaowen TANG
;
Zhengming JIN
;
Miao MIAO
;
Aining SUN
;
Depei WU
Author Information
1. The First Affiliated Hospital of Soochow University, Jiangsu Institute of Hematology, Collaborative Innovation Center of Hematology, Suzhou Institute of Blood and Marrow Transplantation, Suzhou 215006, China
- Publication Type:Journal Article
- Keywords:
Antigen, CD4;
Lymphoma, T cell;
Chimeric antigen receptor modified T cell
- From:
Chinese Journal of Hematology
2018;39(2):148-152
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the specific killing effect of CD4 membrane protein targeted chimeric antigen receptor modified T (CAR-T) cell.
Methods:The second generation CD4 targeted chimeric antigen receptor containing 4-1BB costimulation domain was insert into lentiviral vector through recombinant DNA technology. Lentivirus was prepared and packaged by 293T cells with four plasmids. Beads activated T cells were transduced with lentivirus and the transduction efficiency was checked with Protein L and flow cytometry. T cell subsets and IFN-γ concentrations were detected with probe-tagged antibody and cytometric bead assay.
Results:①The transduction efficiency of activated T cells with prepared lentivirus were 50.0%-70.0%. A subset of CD8+ T cell acquired dim expression of CD4 membrane protein after activation. CD4+T cell and CD8+CD4dim T cell were gradually killed by CD4 targeted CAR-T post lentivirus transduction. ②The kill efficacy of CD4 targeted CAR-T cell and control T cell toward KARPAS 299 T cell at an E∶T ratio of 8∶1 for 24 h was (96.9±2.1)% and (11.2±3.1)%, CAR-T cell has a higher killing efficacy than control T cell (t=7.137, P=0.028). The IFN-γ concentrations in culture supernatant of CAR-T cell with K562-CD4 cell, CAR-T cell with K562 cell and CAR-T cell alone were (15 648±2 168), (1 978±354) and (1 785±268) pg/ml, CAR-T cell cocultured with K562-CD4 cell produced more IFN-γ than the other two controls (P<0.01).
Conclusions:CD4 targeted CAR-T has an immunophenotype of CD8+CD4-T cell. CD4 targeted CAR-T cell has killing efficacy toward normal CD4+T cell and CD4+T lymphoma cell. CD4 targeted CAR-T cell also has a killing efficacy toward CD4dim target cell.
