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GMP-Grade Manufacturing of T Cells Engineered to Express a Defined γδTCR
γ9δ2T cells play a critical role in daily cancer immune surveillance by sensing cancer-mediated metabolic changes. However, a major limitation of the therapeutic application of γ9δ2T cells is their diversity and regulation through innate co-receptors. In order to overcome natural obstacles of γ9δ2T...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5988845/ https://www.ncbi.nlm.nih.gov/pubmed/29899740 http://dx.doi.org/10.3389/fimmu.2018.01062 |
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author | Straetemans, Trudy Kierkels, Guido J. J. Doorn, Ruud Jansen, Koen Heijhuurs, Sabine dos Santos, Joao M. van Muyden, Anna D. D. Vie, Henri Clemenceau, Béatrice Raymakers, Reinier de Witte, Moniek Sebestyén, Zsolt Kuball, Jürgen |
author_facet | Straetemans, Trudy Kierkels, Guido J. J. Doorn, Ruud Jansen, Koen Heijhuurs, Sabine dos Santos, Joao M. van Muyden, Anna D. D. Vie, Henri Clemenceau, Béatrice Raymakers, Reinier de Witte, Moniek Sebestyén, Zsolt Kuball, Jürgen |
author_sort | Straetemans, Trudy |
collection | PubMed |
description | γ9δ2T cells play a critical role in daily cancer immune surveillance by sensing cancer-mediated metabolic changes. However, a major limitation of the therapeutic application of γ9δ2T cells is their diversity and regulation through innate co-receptors. In order to overcome natural obstacles of γ9δ2T cells, we have developed the concept of T cells engineered to express a defined γδT cell receptor (TEGs). This next generation of chimeric antigen receptor engineered T (CAR-T) cells not only allows for targeting of hematological but also of solid tumors and, therefore, overcomes major limitations of many CAR-T and γδT cell strategies. Here, we report on the development of a robust manufacturing procedure of T cells engineered to express the high affinity Vγ9Vδ2T cell receptor (TCR) clone 5 (TEG001). We determined the best concentration of anti-CD3/CD28 activation and expansion beads, optimal virus titer, and cell density for retroviral transduction, and validated a Good Manufacturing Practice (GMP)-grade purification procedure by utilizing the CliniMACS system to deplete non- and poorly-engineered T cells. To the best of our knowledge, we have developed the very first GMP manufacturing procedure in which αβTCR depletion is used as a purification method, thereby delivering untouched clinical grade engineered immune cells. This enrichment method is applicable to any engineered T cell product with a reduced expression of endogenous αβTCRs. We report on release criteria and the stability of TEG001 drug substance and TEG001 drug product. The GMP-grade production procedure is now approved by Dutch authorities and allows TEG001 to be generated in cell numbers sufficient to treat patients within the approved clinical trial NTR6541. NTR6541 will investigate the safety and tolerability of TEG001 in patients with relapsed/refractory acute myeloid leukemia, high-risk myelodysplastic syndrome, and relapsed/refractory multiple myeloma. |
format | Online Article Text |
id | pubmed-5988845 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-59888452018-06-13 GMP-Grade Manufacturing of T Cells Engineered to Express a Defined γδTCR Straetemans, Trudy Kierkels, Guido J. J. Doorn, Ruud Jansen, Koen Heijhuurs, Sabine dos Santos, Joao M. van Muyden, Anna D. D. Vie, Henri Clemenceau, Béatrice Raymakers, Reinier de Witte, Moniek Sebestyén, Zsolt Kuball, Jürgen Front Immunol Immunology γ9δ2T cells play a critical role in daily cancer immune surveillance by sensing cancer-mediated metabolic changes. However, a major limitation of the therapeutic application of γ9δ2T cells is their diversity and regulation through innate co-receptors. In order to overcome natural obstacles of γ9δ2T cells, we have developed the concept of T cells engineered to express a defined γδT cell receptor (TEGs). This next generation of chimeric antigen receptor engineered T (CAR-T) cells not only allows for targeting of hematological but also of solid tumors and, therefore, overcomes major limitations of many CAR-T and γδT cell strategies. Here, we report on the development of a robust manufacturing procedure of T cells engineered to express the high affinity Vγ9Vδ2T cell receptor (TCR) clone 5 (TEG001). We determined the best concentration of anti-CD3/CD28 activation and expansion beads, optimal virus titer, and cell density for retroviral transduction, and validated a Good Manufacturing Practice (GMP)-grade purification procedure by utilizing the CliniMACS system to deplete non- and poorly-engineered T cells. To the best of our knowledge, we have developed the very first GMP manufacturing procedure in which αβTCR depletion is used as a purification method, thereby delivering untouched clinical grade engineered immune cells. This enrichment method is applicable to any engineered T cell product with a reduced expression of endogenous αβTCRs. We report on release criteria and the stability of TEG001 drug substance and TEG001 drug product. The GMP-grade production procedure is now approved by Dutch authorities and allows TEG001 to be generated in cell numbers sufficient to treat patients within the approved clinical trial NTR6541. NTR6541 will investigate the safety and tolerability of TEG001 in patients with relapsed/refractory acute myeloid leukemia, high-risk myelodysplastic syndrome, and relapsed/refractory multiple myeloma. Frontiers Media S.A. 2018-05-30 /pmc/articles/PMC5988845/ /pubmed/29899740 http://dx.doi.org/10.3389/fimmu.2018.01062 Text en Copyright © 2018 Straetemans, Kierkels, Doorn, Jansen, Heijhuurs, dos Santos, van Muyden, Vie, Clemenceau, Raymakers, de Witte, Sebestyén and Kuball. https://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Immunology Straetemans, Trudy Kierkels, Guido J. J. Doorn, Ruud Jansen, Koen Heijhuurs, Sabine dos Santos, Joao M. van Muyden, Anna D. D. Vie, Henri Clemenceau, Béatrice Raymakers, Reinier de Witte, Moniek Sebestyén, Zsolt Kuball, Jürgen GMP-Grade Manufacturing of T Cells Engineered to Express a Defined γδTCR |
title | GMP-Grade Manufacturing of T Cells Engineered to Express a Defined γδTCR |
title_full | GMP-Grade Manufacturing of T Cells Engineered to Express a Defined γδTCR |
title_fullStr | GMP-Grade Manufacturing of T Cells Engineered to Express a Defined γδTCR |
title_full_unstemmed | GMP-Grade Manufacturing of T Cells Engineered to Express a Defined γδTCR |
title_short | GMP-Grade Manufacturing of T Cells Engineered to Express a Defined γδTCR |
title_sort | gmp-grade manufacturing of t cells engineered to express a defined γδtcr |
topic | Immunology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5988845/ https://www.ncbi.nlm.nih.gov/pubmed/29899740 http://dx.doi.org/10.3389/fimmu.2018.01062 |
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