Abstract
Genetic modification of primary mouse T cells with chimeric antigen receptors (CAR) has emerged as an important tool for optimizing adoptive T cell immunotherapy strategies. However, limitations in current protocols for generating highly pure and sufficient numbers of enriched effector and helper CAR+ T cell subsets remain problematic. Here, we describe a new retroviral transduction protocol for successfully generating transduced CD8+ and CD4+ T lymphocytes for in vitro and in vivo characterization.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Pham CD, Mitchell DA (2012) Chasing cancer with chimeric antigen receptor therapy. Immunotherapy 4(4):365–367
Rosenberg SA (2012) Raising the bar: the curative potential of human cancer immunotherapy. Sci Transl Med 4(127):127ps8
Curran KJ et al (2012) Chimeric antigen receptors for T cell immunotherapy: current understanding and future directions. J Gene Med 14(6):405–415
Xie Y et al (2010) Naive tumor-specific CD4+ T cells differentiated in vivo eradicate established melanoma. J Exp Med 207(3):651–667
Moeller M et al (2005) Adoptive transfer of gene-engineered CD4+ helper T cells induces potent primary and secondary tumor rejection. Blood 106(9):2995–3003
Tosolini M et al (2011) Clinical impact of different classes of infiltrating T cytotoxic and helper cells (Th1, Th2, Treg, Th17) in patients with colorectal cancer. Cancer Res 71(4):1263–1271
Moeller M et al (2007) Sustained antigen-specific antitumor recall response mediated by gene-modified CD4+ T helper-1 and CD8+ T cells. Cancer Res 67(23):11428–11437
Ellyard JI et al (2007) Th2-mediated anti-tumour immunity: friend or foe? Tissue Antigens 70(1):1–11
Noelle RJ, Nowak EC (2010) Cellular sources and immune functions of interleukin-9. Nat Rev Immunol 10(10):683–687
Sfanos KS et al (2008) Phenotypic analysis of prostate-infiltrating lymphocytes reveals Th17 and Treg skewing. Clin Cancer Res 14(11):3254–3261
Chen D et al (2012) Increased IL-17-producing CD4+ T cells in patients with esophageal cancer. Cell Immunol 272(2):166–174
Oleinika K et al (2013) Suppression, subversion and escape: the role of regulatory T cells in cancer progression. Clin Exp Immunol 171(1):36–45
Kaplan MH (2013) Th9 cells: differentiation and disease. Immunol Rev 252(1):104–115
Korn T et al (2009) IL-17 and Th17 Cells. Annu Rev Immunol 27(1):485–517
Cheadle EJ et al (2010) Natural expression of the CD19 antigen impacts the long-term engraftment but not antitumor activity of CD19-specific engineered T cells. J Immunol 184(4):1885–1896
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
John, L.B., Chee, T.M., Gilham, D.E., Darcy, P.K. (2014). Genetic Modification of Mouse Effector and Helper T Lymphocytes Expressing a Chimeric Antigen Receptor. In: Lawman, M., Lawman, P. (eds) Cancer Vaccines. Methods in Molecular Biology, vol 1139. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0345-0_16
Download citation
DOI: https://doi.org/10.1007/978-1-4939-0345-0_16
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-0344-3
Online ISBN: 978-1-4939-0345-0
eBook Packages: Springer Protocols