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Generation of Chimeric Antigen Receptor T Cells Using Gammaretroviral Vectors

  • Feiyan Mo
  • Maksim MamonkinEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2086)

Abstract

Manufacturing chimeric antigen receptor (CAR)-modified T cells requires incorporation of the CAR transgene, for which viral vectors are most often used. Here, we describe the generation of CAR T cells using primary human T cells and a non–self-inactivating gammaretroviral vector encoding a CAR transgene. The gammaretroviral vector is produced by 293T cells transiently transfected with DNA plasmids encoding necessary components of the viral vector. The resulting viral particles efficiently infect activated T cells and integrate the CAR transgene into the genome of dividing cells for stable expression.

Key words

Gammaretroviral vectors Vector production CAR T cell generation T cell transduction Chimeric antigen receptors 

Notes

Acknowledgments

The authors thank Catherine Gillespie for editing the chapter. This work was supported by the CPRIT Awards RP180810 and RP150611, and by the Leukemia and Lymphoma Society TRP Award.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Center for Cell and Gene Therapy, Texas Children’s Hospital, Houston Methodist HospitalBaylor College of MedicineHoustonUSA
  2. 2.Interdepartmental Graduate Program in Translational Biology and Molecular MedicineBaylor College of MedicineHoustonUSA
  3. 3.Department of Pathology and ImmunologyBaylor College of MedicineHoustonUSA

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