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 K. Rendahl, Edward A. Berger, Elizabeth Connick, Pamela J. Skinner

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

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

Bibliographical note

Funding Information:
This study was supported by NIH grants 5R01AI096966-06S1 (P.S., E.C., and E.B.), 1UM1AI26617 (P.S., E.C., and E.B.), and P51OD011106 / P51RR000167 (E.R.), MN REACH grant 5U01HL127479-03 (P.S.), 1R01A143380-01 (P.S. and E.B.), and 1UM14126617 (P.S. and E.C.); as well as funds provided by the NIAID Division of Intramural Research and the NIH Intramural AIDS Targeted Antiviral Program . Anti-CD3 and anti-CD28 used in these studies was provided by the NIH Nonhuman Primate Reagent Resource ( R24 OD010976 and U24 AI126683 ). IL-2 used in these studies was provided by The NCI Preclinical Repository.

Funding Information:
This study was supported by NIH grants 5R01AI096966-06S1 (P.S. E.C. and E.B.), 1UM1AI26617 (P.S. E.C. and E.B.), and P51OD011106/P51RR000167 (E.R.), MN REACH grant 5U01HL127479-03 (P.S.), 1R01A143380-01 (P.S. and E.B.), and 1UM14126617 (P.S. and E.C.); as well as funds provided by the NIAID Division of Intramural Research and the NIH Intramural AIDS Targeted Antiviral Program. Anti-CD3 and anti-CD28 used in these studies was provided by the NIH Nonhuman Primate Reagent Resource (R24 OD010976 and U24 AI126683). IL-2 used in these studies was provided by The NCI Preclinical Repository. We thank Chi Phan and Jhomary Alegria-Berrocal at the University of Minnesota for assistance with gammaretroviral production, Andrea Weiler at the University of Wisconsin-Madison for conducting viral load assays, and Kim Weisgrau at the University of Wisconsin-Madison for isolation of rhesus macaque PBMCs. We would also like to thank Dr. Scott McIvor at the University of Minnesota, Dr. Leslie Kean at Harvard Medical School, Dr. Catherine Bollard at the Children's Research Institute, Dr. Christopher Peterson at the Fred Hutchinson Cancer Center, Dr. Matthew Trivett at NCI, Dr. Agne Taraseviciute at Seattle Children's Hospital, and Dr. Conrad Russell Cruz at the Children's Research Institute for their help in developing these methods.

Publisher Copyright:
© 2019 The Author(s)

Keywords

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

PubMed: MeSH publication types

  • Journal Article

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