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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...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
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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. |
format | Online Article Text |
id | pubmed-7730610 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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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