Identification of Potent CD19 scFv for CAR T Cells through scFv Screening with NK/T-Cell Line

被引:13
作者
Kang, Chung Hyo [1 ,2 ]
Kim, Yeongrin [1 ,3 ]
Lee, Heung Kyoung [1 ]
Lee, So Myoung [1 ]
Jeong, Hye Gwang [2 ]
Choi, Sang Un [1 ]
Park, Chi Hoon [1 ,3 ]
机构
[1] Korea Res Inst Chem Technol, Therapeut & Biotechnol Div, POB 107, Daejeon 34114, South Korea
[2] Chungnam Natl Univ, Coll Pharm, Daejeon 34134, South Korea
[3] Korea Univ Sci & Technol, Med Chem & Pharmacol, Daejeon 34113, South Korea
关键词
CD19; chimeric-antigen receptor; scFv; FMC63; leukemia; CHIMERIC ANTIGEN RECEPTOR; NK-92; CELLS; CLINICAL DEVELOPMENT; MONOCLONAL-ANTIBODY; THERAPY; ACTIVATION; LEUKEMIA; DOMAIN;
D O I
10.3390/ijms21239163
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
CD19 is the most promising target for developing chimeric-antigen receptor (CAR) T cells against B-cell leukemic cancer. Currently, two CAR-T-cell products, Kymriah and Yescarta, are approved for leukemia patients, and various anti-CD19 CAR T cells are undergoing clinical trial. Most of these anti-CD19 CAR T cells use FMC63 single-chain variable fragments (scFvs) for binding CD19 expressed on the cancer cell surface. In this study, we screened several known CD19 scFvs for developing anti-CD19 CAR T cells. We used the KHYG-1 NK/T-cell line for screening of CD19 scFvs because it has advantages in terms of cell culture and gene transduction compared to primary T cells. Using our CAR construct backbone, we made anti-CD19 CAR constructs which each had CD19 scFvs including FMC63, B43, 25C1, BLY3, 4G7, HD37, HB12a, and HB12b, then made each anti-CD19 CAR KHYG-1 cells. Interestingly, only FMC63 CAR KHYG-1 and 4G7 CAR KHYG-1 efficiently lysed CD19-positive cell lines. In addition, in Jurkat cell line, only these two CAR Jurkat cell lines secreted IL-2 when co-cultured with CD19-positive cell line, NALM-6. Based on these results, we made FMC63 CAR T cells and 4G7 CAR T cells from PBMC. In in vitro lysis assay, 4G7 CAR T cells lysed CD19-positive cell line as well as FMC63 CAR T cells. In in vivo assay with NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice, 4G7 CAR T cells eradicated NALM-6 as potently as FMC63 CAR T cells. Therefore, we anticipate that 4G7 CAR T cells will show as good a result as FMC63 CAR T cells for B-cell leukemia patients.
引用
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页码:1 / 12
页数:12
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