Abstract
The Chimeric Antigen Receptor (CAR) consists of an antibody-derived targeting domain fused with T-cell signaling domains that, when expressed by a T-cell, endows the T-cell with antigen specificity determined by the targeting domain of the CAR. CARs can potentially redirect the effector functions of a T-cell towards any protein and nonprotein target expressed on the cell surface as long as an antibody or similar targeting domain is available. This strategy thereby avoids the requirement of antigen processing and presentation by the target cell and is applicable to nonclassical T-cell targets like carbohydrates. This circumvention of HLA-restriction means that the CAR T-cell approach can be used as a generic tool broadening the potential of applicability of adoptive T-cell therapy. Proof-of-principle studies focusing upon the investigation of the potency of CAR T-cells have primarily focused upon the genetic modification of human and mouse T-cells for therapy. This chapter focuses upon methods to modify T-cells from both species to generate CAR T-cells for functional testing.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Gross G, Waks T, Eshhar Z (1989) Expression of immunoglobulin-T-cell receptor chimeric molecules as functional receptors with antibody-type specificity. Proc Natl Acad Sci U S A 86:10024–10028
Brenner MK, Heslop HE (2010) Adoptive T cell therapy of cancer. Curr Opin Immunol 22:251–257
Bridgeman JS, Hawkins RE, Hombach AA, Abken H, Gilham DE (2010) Building better chimeric antigen receptors for adoptive T cell therapy. Curr Gene Ther 10:77–90
Morgan RA, Dudley ME, Rosenberg SA (2010) Adoptive cell therapy: genetic modification to redirect effector cell specificity. Cancer J 16:336–341
Movassagh M, Boyer O, Burland MC, Leclercq V, Klatzmann D, Lemoine FM (2000) Retrovirus-mediated gene transfer into T cells: 95 % transduction efficiency without further in vitro selection. Hum Gene Ther 11:1189–1200
Lee J, Sadelain M, Brentjens R (2009) Retroviral transduction of murine primary T lymphocytes. Methods Mol Biol 506:83–96
Riviere I, Gallardo HF, Hagani AB, Sadelain M (2000) Retroviral-mediated gene transfer in primary murine and human T-lymphocytes. Mol Biotechnol 15:133–142
Miller DG, Adam MA, Miller AD (1990) Gene transfer by retrovirus vectors occurs only in cells that are actively replicating at the time of infection. Mol Cell Biol 10:4239–4242
Pollok KE, Hanenberg H, Noblitt TW, Schroeder WL, Kato I, Emanuel D, Williams DA (1998) High-efficiency gene transfer into normal and adenosine deaminase-deficient T lymphocytes is mediated by transduction on recombinant fibronectin fragments. J Virol 72:4882–4892
Le Doux JM, Landazuri N, Yarmush ML, Morgan JR (2001) Complexation of retrovirus with cationic and anionic polymers increases the efficiency of gene transfer. Hum Gene Ther 12:1611–1621
DiGiusto DL, Cooper LJ (2007) Preparing clinical grade Ag-specific T cells for adoptive immunotherapy trials. Cytotherapy 9:613–629
Lamers CH, van Elzakker P, van Steenbergen SC, Sleijfer S, Debets R, Gratama JW (2008) Retronectin-assisted retroviral transduction of primary human T lymphocytes under good manufacturing practice conditions: tissue culture bag critically determines cell yield. Cytotherapy 10:406–416
Ferrand C, Robinet E, Contassot E, Certoux JM, Lim A, Herve P, Tiberghien P (2000) Retrovirus-mediated gene transfer in primary T lymphocytes: influence of the transduction/selection process and of ex vivo expansion on the T cell receptor beta chain hypervariable region repertoire. Hum Gene Ther 11:1151–1164
Sauce D, Bodinier M, Garin M, Petracca B, Tonnelier N, Duperrier A, Melo JV, Apperley JF, Ferrand C, Herve P, Lang F, Tiberghien P, Robinet E (2002) Retrovirus-mediated gene transfer in primary T lymphocytes impairs their anti-Epstein-Barr virus potential through both culture-dependent and selection process-dependent mechanisms. Blood 99:1165–1173
Sauce D, Tonnelier N, Duperrier A, Petracca B, de Carvalho Bittencourt M, Saadi M, Saas P, Ferrand C, Herve P, Tiberghien P, Robinet E (2002) Influence of ex vivo expansion and retrovirus-mediated gene transfer on primary T lymphocyte phenotype and functions. J Hematother Stem Cell Res 11:929–940
Gilham DE, Lie ALM, Taylor N, Hawkins RE (2010) Cytokine stimulation and the choice of promoter are critical factors for the efficient transduction of mouse T cells with HIV-1 vectors. J Gene Med 12:129–136
Swainson L, Mongellaz C, Adjali O, Vicente R, Taylor N (2008) Lentiviral transduction of immune cells. Methods Mol Biol 415:301–320
Jensen MC, Clarke P, Tan G, Wright C, Chung-Chang W, Clark TN, Zhang F, Slovak ML, Wu AM, Forman SJ, Raubitschek A (2000) Human T lymphocyte genetic modification with naked DNA. Mol Ther 1:49–55
Birkholz K, Hombach A, Krug C, Reuter S, Kershaw M, Kampgen E, Schuler G, Abken H, Schaft N, Dorrie J (2009) Transfer of mRNA encoding recombinant immunoreceptors reprograms CD4+ and CD8+ T cells for use in the adoptive immunotherapy of cancer. Gene Ther 16:596–604
Zhao Y, Moon E, Carpenito C, Paulos CM, Liu X, Brennan AL, Chew A, Carroll RG, Scholler J, Levine BL, Albelda SM, June CH (2010) Multiple injections of electroporated autologous T cells expressing a chimeric antigen receptor mediate regression of human disseminated tumor. Cancer Res 70:9053–9061
Kruschinski A, Moosmann A, Poschke I, Norell H, Chmielewski M, Seliger B, Kiessling R, Blankenstein T, Abken H, Charo J (2008) Engineering antigen-specific primary human NK cells against HER-2 positive carcinomas. Proc Natl Acad Sci U S A 105:17481–17486
Pegram HJ, Kershaw MH, Darcy PK (2009) Genetic modification of natural killer cells for adoptive cellular immunotherapy. Immunotherapy 1:623–630
Biglari A, Southgate TD, Fairbairn LJ, Gilham DE (2006) Human monocytes expressing a CEA-specific chimeric CD64 receptor specifically target CEA-expressing tumour cells in vitro and in vivo. Gene Ther 13:602–610
Pouw NM, Westerlaken EJ, Willemsen RA, Debets R (2007) Gene transfer of human TCR in primary murine T cells is improved by pseudo-typing with amphotropic and ecotropic envelopes. J Gene Med 9:561–570
Boczkowski D, Nair SK, Nam JH, Lyerly HK, Gilboa E (2000) Induction of tumor immunity and cytotoxic T lymphocyte responses using dendritic cells transfected with messenger RNA amplified from tumor cells. Cancer Res 60:1028–1034
Weijtens ME, Willemsen RA, Hart EH, Bolhuis RL (1998) A retroviral vector system ‘STITCH’ in combination with an optimized single chain antibody chimeric receptor gene structure allows efficient gene transduction and expression in human T lymphocytes. Gene Ther 5:1195–1203
Fehse B, Kustikova OS, Li Z, Wahlers A, Bohn W, Beyer WR, Chalmers D, Tiberghien P, Kuhlcke K, Zander AR, Baum C (2002) A novel ‘sort-suicide’ fusion gene vector for T cell manipulation. Gene Ther 9:1633–1638
Swift S, Lorens J, Achacoso P, Nolan GP (2001) Rapid production of retroviruses for efficient gene delivery to mammalian cells using 293 T cell-based systems. Curr Protoc Immunol 10:14–29, Chapter 10, Unit 10 17 C
Miller AD, Garcia JV, von Suhr N, Lynch CM, Wilson C, Eiden MV (1991) Construction and properties of retrovirus packaging cells based on gibbon ape leukemia virus. J Virol 65:2220–2224
Lamers CH, van Elzakker P, Luider BA, van Steenbergen SC, Sleijfer S, Debets R, Gratama JW (2008) Retroviral vectors for clinical immunogene therapy are stable for up to 9 years. Cancer Gene Ther 15:268–274
Lamers CH, Willemsen R, van Elzakker P, van Steenbergen-Langeveld S, Broertjes M, Oosterwijk-Wakka J, Oosterwijk E, Sleijfer S, Debets R, Gratama JW (2011) Immune responses to transgene and retroviral vector in patients treated with ex vivo-engineered T cells. Blood 117:72–82
Schroten C, Kraaij R, Veldhoven JL, Berrevoets CA, den Bakker MA, Ma Q, Sadelain M, Bangma CH, Willemsen RA, Debets R (2010) T cell activation upon exposure to patient-derived tumor tissue: a functional assay to select patients for adoptive T cell therapy. J Immunol Methods 359:11–20
Lamers CH, Willemsen RA, Luider BA, Debets R, Bolhuis RL (2002) Protocol for gene transduction and expansion of human T lymphocytes for clinical immunogene therapy of cancer. Cancer Gene Ther 9:613–623
Hollyman D, Stefanski J, Przybylowski M, Bartido S, Borquez-Ojeda O, Taylor C, Yeh R, Capacio V, Olszewska M, Hosey J, Sadelain M, Brentjens RJ, Riviere I (2009) Manufacturing validation of biologically functional T cells targeted to CD19 antigen for autologous adoptive cell therapy. J Immunother 32:169–180
Pouw N, Treffers-Westerlaken E, Kraan J, Wittink F, ten Hagen T, Verweij J, Debets R (2010) Combination of IL-21 and IL-15 enhances tumour-specific cytotoxicity and cytokine production of TCR-transduced primary T cells. Cancer Immunol Immunother 59:921–931
Suhoski MM, Golovina TN, Aqui NA, Tai VC, Varela-Rohena A, Milone MC, Carroll RG, Riley JL, June CH (2007) Engineering artificial antigen-presenting cells to express a diverse array of co-stimulatory molecules. Mol Ther 15:981–988
Pouw N, Treffers-Westerlaken E, Mondino A, Lamers C, Debets R (2010) TCR gene-engineered T cell: limited T cell activation and combined use of IL-15 and IL-21 ensure minimal differentiation and maximal antigen-specificity. Mol Immunol 47:1411–1420
Acknowledgments
The work has been supported by the European Union FP6 program “ATTACK” and FP7 Network “ATTRACT.”
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Cheadle, E.J. et al. (2012). Chimeric Antigen Receptors for T-Cell Based Therapy. In: Chames, P. (eds) Antibody Engineering. Methods in Molecular Biology, vol 907. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-974-7_36
Download citation
DOI: https://doi.org/10.1007/978-1-61779-974-7_36
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-973-0
Online ISBN: 978-1-61779-974-7
eBook Packages: Springer Protocols