Transduction and expansion of primary T cells in nine days with maintenance of central memory phenotype

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

Emerging immunotherapies to treat infectious diseases and cancers often involve transduction of cellular populations with genes encoding disease-targeting proteins. For example, chimeric antigen receptor (CAR)-T cells to treat cancers and viral infections involve the transduction of T cells with synthetic genes encoding CAR molecules. The CAR molecules make the T cells specifically recognize and kill cancer or virally infected cells. Cells can also be co-transduced with other genes of interest. For example, cells can be co-transduced with genes encoding proteins that target cells to specific locations. Here, we present a protocol to transduce primary peripheral blood mononuclear cells (PBMCs) with genes encoding a virus-specific CAR and the B cell follicle homing molecule chemokine receptor type 5 (CXCR5). This procedure takes nine days and results in transduced T cell populations that maintain a central memory phenotype. Maintenance of a central memory or less differentiated phenotype has been shown to associate with persistence of cells post-infusion. Furthermore, cells produced with this method show high levels of viability, high levels of co-expression of the two transduced genes, and large enough quantities of cells for immunotherapeutic infusion. This nine-day protocol may be broadly used for CAR-T cell and other T cell immunotherapy approaches. The methods described here are based on studies presented in our previous publications.

Original languageEnglish (US)
Article numbere60400
JournalJournal of Visualized Experiments
Volume2020
Issue number157
DOIs
StatePublished - Mar 2020

Bibliographical note

Publisher Copyright:
© 2020 Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License.

Keywords

  • Bioengineering
  • CAR-T cells
  • Central memory
  • Expansion
  • Issue 157
  • PBMC
  • Retrovirus
  • Transduction

Fingerprint

Dive into the research topics of 'Transduction and expansion of primary T cells in nine days with maintenance of central memory phenotype'. Together they form a unique fingerprint.

Cite this