Abstract
Adoptive T cell therapy has demonstrated tremendous outcomes against treatment-refractory leukemias and solid tumor malignancies. As opposed to industry-developed drugs that are manufactured and dispensed to hospitals and/or patients, T cells are produced in academic laboratories for clinical research and are a highly personalized therapy that represents a “living drug.” The technology behind genetic modification of primary T cells has been developed and refined by a few academic medical centers. We anticipate that the exciting results generated by these efforts will lead to further investigation by other academic and industry institutions. To facilitate this adaptation we present optimized protocols for gammaretroviral production, T cell isolation, and genetic modification to create gene-targeted T cells.
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References
Davila ML, Riviere I, Wang X et al (2014) Efficacy and toxicity management of 19-28z CAR T cell therapy in B cell acute lymphoblastic leukemia. Sci Transl Med 6:224ra225. doi:10.1126/scitranslmed.3008226
Stevanovic S, Draper LM, Langhan MM et al (2015) Complete regression of metastatic cervical cancer after treatment with human papillomavirus-targeted tumor-infiltrating T cells. J Clin Oncol 33(14):1543–1550. doi:10.1200/JCO.2014.58.9093
Hodi FS, O’Day SJ, McDermott DF et al (2010) Improved survival with ipilimumab in patients with metastatic melanoma. N Engl J Med 363(8):711–723. doi:10.1056/NEJMoa1003466
Topp MS, Gokbuget N, Zugmaier G et al (2014) Phase II trial of the anti-CD19 bispecific T cell-engager blinatumomab shows hematologic and molecular remissions in patients with relapsed or refractory b-precursor acute lymphoblastic leukemia. J Clin Oncol 32(36):4134–4140. doi:10.1200/JCO.2014.56.3247
Restifo NP, Dudley ME, Rosenberg SA (2012) Adoptive immunotherapy for cancer: harnessing the T cell response. Nat Rev Immunol 12(4):269–281. doi:10.1038/nri3191
Sadelain M, Riviere I, Brentjens R (2003) Targeting tumours with genetically enhanced T lymphocytes. Nat Rev Cancer 3(1):35–45. doi:10.1038/nrc971
Davila ML, Brentjens R, Wang X et al (2012) How do CARs work?: Early insights from recent clinical studies targeting CD19. Oncoimmunology 1(9):1577–1583. doi:10.4161/onci.22524
Lee DW, Kochenderfer JN, Stetler-Stevenson M et al (2014) T cells expressing CD19 chimeric antigen receptors for acute lymphoblastic leukaemia in children and young adults: a phase 1 dose-escalation trial. Lancet. doi:10.1016/S0140-6736(14)61403-3
Hollyman D, Stefanski J, Przybylowski M et al (2009) Manufacturing validation of biologically functional T cells targeted to CD19 antigen for autologous adoptive cell therapy. J Immunother 32(2):169–180. doi:10.1097/CJI.0b013e318194a6e8
Kochenderfer JN, Feldman SA, Zhao Y et al (2009) Construction and preclinical evaluation of an anti-CD19 chimeric antigen receptor. J Immunother 32(7):689–702. doi:10.1097/CJI.0b013e3181ac6138
Lee J, Sadelain M, Brentjens R (2009) Retroviral transduction of murine primary T lymphocytes. Methods Mol Biol 506:83–96. doi:10.1007/978-1-59745-409-4_7
Zheng Z, Chinnasamy N, Morgan RA (2012) Protein L: a novel reagent for the detection of chimeric antigen receptor (CAR) expression by flow cytometry. J Transl Med 10:29. doi:10.1186/1479-5876-10-29
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Li, G., Park, K., Davila, M.L. (2017). Gammaretroviral Production and T Cell Transduction to Genetically Retarget Primary T Cells Against Cancer. In: Lugli, E. (eds) T-Cell Differentiation. Methods in Molecular Biology, vol 1514. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6548-9_9
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DOI: https://doi.org/10.1007/978-1-4939-6548-9_9
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Online ISBN: 978-1-4939-6548-9
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