Rapid Transduction and Expansion of Transduced T Cells with Maintenance of Central Memory Populations

Mary S Pampusch, Kumudhini Preethi Haran, Geoffrey T. Hart, Eva G. Rakasz, Aaron Rendahl, Edward A. Berger, Elizabeth Connick, Pamela J Skinner

Research output: Contribution to journalArticle

Abstract

Chimeric antigen receptor (CAR)-T cells show great promise in treating cancers and viral infections. However, most protocols developed to expand T cells require relatively long periods of time in culture, potentially leading to progression toward populations of terminally differentiated effector memory cells. Here, we describe in detail a 9-day protocol for CAR gene transduction and expansion of primary rhesus macaque peripheral blood mononuclear cells (PBMCs). Cells produced and expanded with this method show high levels of viability, high levels of co-expression of two transduced genes, retention of the central memory phenotype, and sufficient quantity for immunotherapeutic infusion of 1–2 × 108 cells/kg in a 10 kg rhesus macaque. This 9-day protocol may be broadly used for CAR-T cell and other T cell immunotherapy approaches to decrease culture time and increase maintenance of central memory populations.

Original languageEnglish (US)
Pages (from-to)1-10
Number of pages10
JournalMolecular Therapy - Methods and Clinical Development
Volume16
DOIs
StatePublished - Mar 13 2020

Fingerprint

Maintenance
T-Cell Antigen Receptor
Macaca mulatta
T-Lymphocytes
Population
Antigen Receptors
Virus Diseases
Immunotherapy
Genes
Blood Cells
Phenotype
Neoplasms

Keywords

  • CAR-T cells
  • PBMC
  • central memory
  • expansion
  • retrovirus
  • rhesus macaque
  • transduction

Cite this

Rapid Transduction and Expansion of Transduced T Cells with Maintenance of Central Memory Populations. / Pampusch, Mary S; Haran, Kumudhini Preethi; Hart, Geoffrey T.; Rakasz, Eva G.; Rendahl, Aaron; Berger, Edward A.; Connick, Elizabeth; Skinner, Pamela J.

In: Molecular Therapy - Methods and Clinical Development, Vol. 16, 13.03.2020, p. 1-10.

Research output: Contribution to journalArticle

Pampusch, Mary S ; Haran, Kumudhini Preethi ; Hart, Geoffrey T. ; Rakasz, Eva G. ; Rendahl, Aaron ; Berger, Edward A. ; Connick, Elizabeth ; Skinner, Pamela J. / Rapid Transduction and Expansion of Transduced T Cells with Maintenance of Central Memory Populations. In: Molecular Therapy - Methods and Clinical Development. 2020 ; Vol. 16. pp. 1-10.
@article{130662ba53664dbaa9eaf729c040d68b,
title = "Rapid Transduction and Expansion of Transduced T Cells with Maintenance of Central Memory Populations",
abstract = "Chimeric antigen receptor (CAR)-T cells show great promise in treating cancers and viral infections. However, most protocols developed to expand T cells require relatively long periods of time in culture, potentially leading to progression toward populations of terminally differentiated effector memory cells. Here, we describe in detail a 9-day protocol for CAR gene transduction and expansion of primary rhesus macaque peripheral blood mononuclear cells (PBMCs). Cells produced and expanded with this method show high levels of viability, high levels of co-expression of two transduced genes, retention of the central memory phenotype, and sufficient quantity for immunotherapeutic infusion of 1–2 × 108 cells/kg in a 10 kg rhesus macaque. This 9-day protocol may be broadly used for CAR-T cell and other T cell immunotherapy approaches to decrease culture time and increase maintenance of central memory populations.",
keywords = "CAR-T cells, PBMC, central memory, expansion, retrovirus, rhesus macaque, transduction",
author = "Pampusch, {Mary S} and Haran, {Kumudhini Preethi} and Hart, {Geoffrey T.} and Rakasz, {Eva G.} and Aaron Rendahl and Berger, {Edward A.} and Elizabeth Connick and Skinner, {Pamela J}",
year = "2020",
month = "3",
day = "13",
doi = "10.1016/j.omtm.2019.09.007",
language = "English (US)",
volume = "16",
pages = "1--10",
journal = "Molecular Therapy - Methods and Clinical Development",
issn = "2329-0501",
publisher = "Nature Publishing Group",

}

TY - JOUR

T1 - Rapid Transduction and Expansion of Transduced T Cells with Maintenance of Central Memory Populations

AU - Pampusch, Mary S

AU - Haran, Kumudhini Preethi

AU - Hart, Geoffrey T.

AU - Rakasz, Eva G.

AU - Rendahl, Aaron

AU - Berger, Edward A.

AU - Connick, Elizabeth

AU - Skinner, Pamela J

PY - 2020/3/13

Y1 - 2020/3/13

N2 - Chimeric antigen receptor (CAR)-T cells show great promise in treating cancers and viral infections. However, most protocols developed to expand T cells require relatively long periods of time in culture, potentially leading to progression toward populations of terminally differentiated effector memory cells. Here, we describe in detail a 9-day protocol for CAR gene transduction and expansion of primary rhesus macaque peripheral blood mononuclear cells (PBMCs). Cells produced and expanded with this method show high levels of viability, high levels of co-expression of two transduced genes, retention of the central memory phenotype, and sufficient quantity for immunotherapeutic infusion of 1–2 × 108 cells/kg in a 10 kg rhesus macaque. This 9-day protocol may be broadly used for CAR-T cell and other T cell immunotherapy approaches to decrease culture time and increase maintenance of central memory populations.

AB - Chimeric antigen receptor (CAR)-T cells show great promise in treating cancers and viral infections. However, most protocols developed to expand T cells require relatively long periods of time in culture, potentially leading to progression toward populations of terminally differentiated effector memory cells. Here, we describe in detail a 9-day protocol for CAR gene transduction and expansion of primary rhesus macaque peripheral blood mononuclear cells (PBMCs). Cells produced and expanded with this method show high levels of viability, high levels of co-expression of two transduced genes, retention of the central memory phenotype, and sufficient quantity for immunotherapeutic infusion of 1–2 × 108 cells/kg in a 10 kg rhesus macaque. This 9-day protocol may be broadly used for CAR-T cell and other T cell immunotherapy approaches to decrease culture time and increase maintenance of central memory populations.

KW - CAR-T cells

KW - PBMC

KW - central memory

KW - expansion

KW - retrovirus

KW - rhesus macaque

KW - transduction

UR - http://www.scopus.com/inward/record.url?scp=85073101687&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85073101687&partnerID=8YFLogxK

U2 - 10.1016/j.omtm.2019.09.007

DO - 10.1016/j.omtm.2019.09.007

M3 - Article

AN - SCOPUS:85073101687

VL - 16

SP - 1

EP - 10

JO - Molecular Therapy - Methods and Clinical Development

JF - Molecular Therapy - Methods and Clinical Development

SN - 2329-0501

ER -