Expansion of Human Tregs from Cryopreserved Umbilical Cord Blood for GMP-Compliant Autologous Adoptive Cell Transfer Therapy
Umbilical cord blood is a traditional and convenient source of cells for hematopoietic stem cell transplantation. Thymic regulatory T cells (Tregs) are also present in cord blood, and there is growing interest in the use of autologous Tregs to provide a low-risk, fully human leukocyte antigen (HLA)-...
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Elsevier
2017-03-01
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Series: | Molecular Therapy: Methods & Clinical Development |
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Online Access: | http://www.sciencedirect.com/science/article/pii/S232905011630136X |
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record_format |
Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Howard R. Seay Amy L. Putnam Judit Cserny Amanda L. Posgai Emma H. Rosenau John R. Wingard Kate F. Girard Morey Kraus Angela P. Lares Heather L. Brown Katherine S. Brown Kristi T. Balavage Leeana D. Peters Ashley N. Bushdorf Mark A. Atkinson Jeffrey A. Bluestone Michael J. Haller Todd M. Brusko |
spellingShingle |
Howard R. Seay Amy L. Putnam Judit Cserny Amanda L. Posgai Emma H. Rosenau John R. Wingard Kate F. Girard Morey Kraus Angela P. Lares Heather L. Brown Katherine S. Brown Kristi T. Balavage Leeana D. Peters Ashley N. Bushdorf Mark A. Atkinson Jeffrey A. Bluestone Michael J. Haller Todd M. Brusko Expansion of Human Tregs from Cryopreserved Umbilical Cord Blood for GMP-Compliant Autologous Adoptive Cell Transfer Therapy Molecular Therapy: Methods & Clinical Development autoimmunity cord blood adoptive cell transfer regulatory T cell type 1 diabetes T cell receptor good manufacturing practices |
author_facet |
Howard R. Seay Amy L. Putnam Judit Cserny Amanda L. Posgai Emma H. Rosenau John R. Wingard Kate F. Girard Morey Kraus Angela P. Lares Heather L. Brown Katherine S. Brown Kristi T. Balavage Leeana D. Peters Ashley N. Bushdorf Mark A. Atkinson Jeffrey A. Bluestone Michael J. Haller Todd M. Brusko |
author_sort |
Howard R. Seay |
title |
Expansion of Human Tregs from Cryopreserved Umbilical Cord Blood for GMP-Compliant Autologous Adoptive Cell Transfer Therapy |
title_short |
Expansion of Human Tregs from Cryopreserved Umbilical Cord Blood for GMP-Compliant Autologous Adoptive Cell Transfer Therapy |
title_full |
Expansion of Human Tregs from Cryopreserved Umbilical Cord Blood for GMP-Compliant Autologous Adoptive Cell Transfer Therapy |
title_fullStr |
Expansion of Human Tregs from Cryopreserved Umbilical Cord Blood for GMP-Compliant Autologous Adoptive Cell Transfer Therapy |
title_full_unstemmed |
Expansion of Human Tregs from Cryopreserved Umbilical Cord Blood for GMP-Compliant Autologous Adoptive Cell Transfer Therapy |
title_sort |
expansion of human tregs from cryopreserved umbilical cord blood for gmp-compliant autologous adoptive cell transfer therapy |
publisher |
Elsevier |
series |
Molecular Therapy: Methods & Clinical Development |
issn |
2329-0501 |
publishDate |
2017-03-01 |
description |
Umbilical cord blood is a traditional and convenient source of cells for hematopoietic stem cell transplantation. Thymic regulatory T cells (Tregs) are also present in cord blood, and there is growing interest in the use of autologous Tregs to provide a low-risk, fully human leukocyte antigen (HLA)-matched cell product for treating autoimmune diseases, such as type 1 diabetes. Here, we describe a good manufacturing practice (GMP)-compatible Treg expansion protocol using fluorescence-activated cell sorting, resulting in a mean 2,092-fold expansion of Tregs over a 16-day culture for a median yield of 1.26 × 109 Tregs from single-donor cryopreserved units. The resulting Tregs passed prior clinical trial release criteria for Treg purity and sterility, including additional rigorous assessments of FOXP3 and Helios expression and epigenetic analysis of the FOXP3 Treg-specific demethylated region (TSDR). Compared with expanded adult peripheral blood Tregs, expanded cord blood Tregs remained more naive, as assessed by continued expression of CD45RA, produced reduced IFN-γ following activation, and effectively inhibited responder T cell proliferation. Immunosequencing of the T cell receptor revealed a remarkably diverse receptor repertoire within cord blood Tregs that was maintained following in vitro expansion. These data support the feasibility of generating GMP-compliant Tregs from cord blood for adoptive cell transfer therapies and highlight potential advantages in terms of safety, phenotypic stability, autoantigen specificity, and tissue distribution. |
topic |
autoimmunity cord blood adoptive cell transfer regulatory T cell type 1 diabetes T cell receptor good manufacturing practices |
url |
http://www.sciencedirect.com/science/article/pii/S232905011630136X |
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doaj-b5555cc4f2ea4d9793b448e24da450ff2020-11-24T21:01:39ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012017-03-014C17819110.1016/j.omtm.2016.12.003Expansion of Human Tregs from Cryopreserved Umbilical Cord Blood for GMP-Compliant Autologous Adoptive Cell Transfer TherapyHoward R. Seay0Amy L. Putnam1Judit Cserny2Amanda L. Posgai3Emma H. Rosenau4John R. Wingard5Kate F. Girard6Morey Kraus7Angela P. Lares8Heather L. Brown9Katherine S. Brown10Kristi T. Balavage11Leeana D. Peters12Ashley N. Bushdorf13Mark A. Atkinson14Jeffrey A. Bluestone15Michael J. Haller16Todd M. Brusko17Department of Pathology, Immunology and Laboratory Medicine, University of Florida College of Medicine, Gainesville, FL 32610, USADiabetes Center and Department of Medicine, University of California, San Francisco, San Francisco, CA 94143, USADepartment of Pathology, Immunology and Laboratory Medicine, University of Florida College of Medicine, Gainesville, FL 32610, USADepartment of Pathology, Immunology and Laboratory Medicine, University of Florida College of Medicine, Gainesville, FL 32610, USADivision of Hematology and Oncology, Department of Medicine, University of Florida College of Medicine, Gainesville, FL 32610, USADivision of Hematology and Oncology, Department of Medicine, University of Florida College of Medicine, Gainesville, FL 32610, USAViaCord, LLC, Waltham, MA 02451, USAViaCord, LLC, Waltham, MA 02451, USADiabetes Center and Department of Medicine, University of California, San Francisco, San Francisco, CA 94143, USACbr Systems, Inc., San Bruno, CA 94066, USACbr Systems, Inc., San Bruno, CA 94066, USADepartment of Pathology, Immunology and Laboratory Medicine, University of Florida College of Medicine, Gainesville, FL 32610, USADepartment of Pathology, Immunology and Laboratory Medicine, University of Florida College of Medicine, Gainesville, FL 32610, USADepartment of Pathology, Immunology and Laboratory Medicine, University of Florida College of Medicine, Gainesville, FL 32610, USADepartment of Pathology, Immunology and Laboratory Medicine, University of Florida College of Medicine, Gainesville, FL 32610, USADiabetes Center and Department of Medicine, University of California, San Francisco, San Francisco, CA 94143, USADepartment of Pediatrics, University of Florida College of Medicine, Gainesville, FL 32610, USADepartment of Pathology, Immunology and Laboratory Medicine, University of Florida College of Medicine, Gainesville, FL 32610, USAUmbilical cord blood is a traditional and convenient source of cells for hematopoietic stem cell transplantation. Thymic regulatory T cells (Tregs) are also present in cord blood, and there is growing interest in the use of autologous Tregs to provide a low-risk, fully human leukocyte antigen (HLA)-matched cell product for treating autoimmune diseases, such as type 1 diabetes. Here, we describe a good manufacturing practice (GMP)-compatible Treg expansion protocol using fluorescence-activated cell sorting, resulting in a mean 2,092-fold expansion of Tregs over a 16-day culture for a median yield of 1.26 × 109 Tregs from single-donor cryopreserved units. The resulting Tregs passed prior clinical trial release criteria for Treg purity and sterility, including additional rigorous assessments of FOXP3 and Helios expression and epigenetic analysis of the FOXP3 Treg-specific demethylated region (TSDR). Compared with expanded adult peripheral blood Tregs, expanded cord blood Tregs remained more naive, as assessed by continued expression of CD45RA, produced reduced IFN-γ following activation, and effectively inhibited responder T cell proliferation. Immunosequencing of the T cell receptor revealed a remarkably diverse receptor repertoire within cord blood Tregs that was maintained following in vitro expansion. These data support the feasibility of generating GMP-compliant Tregs from cord blood for adoptive cell transfer therapies and highlight potential advantages in terms of safety, phenotypic stability, autoantigen specificity, and tissue distribution.http://www.sciencedirect.com/science/article/pii/S232905011630136Xautoimmunitycord bloodadoptive cell transferregulatory T celltype 1 diabetesT cell receptorgood manufacturing practices |