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

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-C...

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Autores principales: Kang, Chung Hyo, Kim, Yeongrin, Lee, Heung Kyoung, Lee, So Myoung, Jeong, Hye Gwang, Choi, Sang Un, Park, Chi Hoon
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7730610/
https://www.ncbi.nlm.nih.gov/pubmed/33271901
http://dx.doi.org/10.3390/ijms21239163
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author Kang, Chung Hyo
Kim, Yeongrin
Lee, Heung Kyoung
Lee, So Myoung
Jeong, Hye Gwang
Choi, Sang Un
Park, Chi Hoon
author_facet Kang, Chung Hyo
Kim, Yeongrin
Lee, Heung Kyoung
Lee, So Myoung
Jeong, Hye Gwang
Choi, Sang Un
Park, Chi Hoon
author_sort Kang, Chung Hyo
collection PubMed
description 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.
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spelling pubmed-77306102020-12-12 Identification of Potent CD19 scFv for CAR T Cells through scFv Screening with NK/T-Cell Line Kang, Chung Hyo Kim, Yeongrin Lee, Heung Kyoung Lee, So Myoung Jeong, Hye Gwang Choi, Sang Un Park, Chi Hoon Int J Mol Sci Article 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. MDPI 2020-12-01 /pmc/articles/PMC7730610/ /pubmed/33271901 http://dx.doi.org/10.3390/ijms21239163 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kang, Chung Hyo
Kim, Yeongrin
Lee, Heung Kyoung
Lee, So Myoung
Jeong, Hye Gwang
Choi, Sang Un
Park, Chi Hoon
Identification of Potent CD19 scFv for CAR T Cells through scFv Screening with NK/T-Cell Line
title Identification of Potent CD19 scFv for CAR T Cells through scFv Screening with NK/T-Cell Line
title_full Identification of Potent CD19 scFv for CAR T Cells through scFv Screening with NK/T-Cell Line
title_fullStr Identification of Potent CD19 scFv for CAR T Cells through scFv Screening with NK/T-Cell Line
title_full_unstemmed Identification of Potent CD19 scFv for CAR T Cells through scFv Screening with NK/T-Cell Line
title_short Identification of Potent CD19 scFv for CAR T Cells through scFv Screening with NK/T-Cell Line
title_sort identification of potent cd19 scfv for car t cells through scfv screening with nk/t-cell line
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7730610/
https://www.ncbi.nlm.nih.gov/pubmed/33271901
http://dx.doi.org/10.3390/ijms21239163
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