Abstract
Genetic modification of tumor-infiltrating lymphocytes (TILs) or circulating T cells has become an important avenue in cancer therapy. Here we describe a comprehensive method for establishing and expanding TIL cultures and genetically modifying them with a gene of interest (GOI) via retroviral transduction or mRNA transfection. The method includes all the important steps starting with TIL extraction from tumors through to the maintenance of the genetically modified TILs. The protocol includes instructions for retroviral transduction and mRNA transfection of circulating T cells or T-cell lines. The GOIs most commonly introduced into the target cells are chimeric antigen receptors (CARs); genetic adjuvants, such as membrane-bound interleukins; and antitumor T-cell receptors (TCRs).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Dudley ME, Wunderlich JR, Shelton TE et al (2003) Generation of tumor-infiltrating lymphocyte cultures for use in adoptive transfer therapy for melanoma patients. J Immunother 26:332–342
Dudley ME, Gross CA, Langhan MM et al (2010) C D 8 + enriched “young” tumor infiltrating lymphocytes can mediate regression of metastatic melanoma. Clin Cancer Res 16(24):6122–6131
Weinstein-Marom H, Gross G, Levi M, Brayer H, Schachter J, Itzhaki O, Besser MJ (2021) Genetic modification of tumor- infiltrating lymphocytes via retroviral transduction. Front Immunol 11:584148
Acknowledgments
We would like to thank Dr. Mikael Schwarz (PhD) for his essential help with the writing and editing of the manuscript. We thank Dr. Gillian Kay (PhD) for fine-tuning the final manuscript. LN is supported by the Israel Science Foundation (ISF), the Israel Cancer Association (ICA), and the State of Israel Ministry of Innovation, Science and Technology Science Forefront grants. LN has direct financial interests in biotechnology companies in the field of cancer immunotherapy.
All figures were created with Biorender.com.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Weinstein-Marom, H. et al. (2024). Genetic Modification of Tumor-Infiltrating Lymphocytes, Peripheral T Cells, and T-Cell Model Cell Lines. In: Siciliano, V., Ceroni, F. (eds) Cancer Immunotherapy. Methods in Molecular Biology, vol 2748. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3593-3_13
Download citation
DOI: https://doi.org/10.1007/978-1-0716-3593-3_13
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-3592-6
Online ISBN: 978-1-0716-3593-3
eBook Packages: Springer Protocols