Review Article | Published:

Targeting tumours with genetically enhanced T lymphocytes


The genetic modification of T lymphocytes is an important approach to investigating normal T-cell biology and to increasing antitumour immunity. A number of genetic strategies aim to increase the recognition of tumour antigens, enhance antitumour activities and prevent T-cell malfunction. T cells can also be engineered to increase safety, as well as to express markers that can be tracked by non-invasive imaging technologies. Genetically modified T cells are therefore proving to be of great value for basic immunology and experimental immunotherapy.

Key Points

  • The genetic modification of T lymphocytes is the basis for novel approaches to study tumour immunity.

  • Antigen specificity can be redirected by the transduction of physiological and chimeric antigen receptors into T cells.

  • T-cell proliferation and function can be increased by genetically manipulating cells to express modified co-stimulatory receptors.

  • T cells can be genetically protected from immunosuppressive factors such as transforming growth factor-β.

  • T cells can also be genetically modified to include regulatable suicide mechanisms, thereby ensuring the safety of adoptive-cell therapy.

  • Expressing constitutive or inducible markers in T cells allows the tracking of cell migration and activation using non-invasive imaging technologies.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


  1. 1

    Houghton, A. N. Cancer antigens: immune recognition of self and altered self. J. Exp. Med. 180, 1–4 (1994).

  2. 2

    Boon, T. & Old, L. J. Cancer tumor antigens. Curr. Opin. Immunol. 9, 681–683 (1997).

  3. 3

    Rosenberg, S. A. Progress in human tumour immunology and immunotherapy. Nature 411, 380–384 (2001).

  4. 4

    Englehard, V. H. et al. Antigens derived from melanocyte differentiation proteins: self-tolerance, autoimmunity, and use for cancer immunotherapy. Immunol. Rev. 188, 136–146 (2002).

  5. 5

    Greenberg, P. D. Adoptive T cell therapy of tumors: mechanisms operative in the recognition and elimination of tumor cells. Adv. Immunol. 49, 281–355 (1991).

  6. 6

    Mavilio, F. et al. Peripheral blood lymphocytes as target cells of retroviral vector-mediated gene transfer. Blood 83, 1988–1997 (1994).

  7. 7

    Bunnell, B. A., Muul, L. M., Donahue, R. E., Blaese, R. M. & Morgan, R. A. High-efficiency retroviral-mediated gene transfer into human and nonhuman primate peripheral blood lymphocytes. Proc. Natl Acad. Sci. USA 92, 7739–7743 (1995).

  8. 8

    Gallardo, H. F., Tan, C., Ory, D. & Sadelain, M. Recombinant retroviruses pseudotyped with the vesicular stomatitis virus G glycoprotein mediate both stable gene transfer and pseudotransduction in human peripheral blood lymphocytes. Blood 90, 952–957 (1997).

  9. 9

    Hagani, A. B., Riviere, I., Tan, C., Krause, A. & Sadelain, M. Activation conditions determine susceptibility of murine primary T-lymphocytes to retroviral infection. J. Gene Med. 1, 341–351 (1999).

  10. 10

    Riviere, I., Gallardo, H. F., Hagani, A. B. & Sadelain, M. Retroviral-mediated gene transfer in primary murine and human T-lymphocytes. Mol. Biotechnol. 15, 133–142 (2000).

  11. 11

    Shibagaki, N., Hanada, K., Yamashita, H., Shimada, S. & Hamada, H. Overexpression of CD82 on human T cells enhances LFA-1/ICAM-1-mediated cell-cell adhesion: functional association between CD82 and LFA-1 in T cell activation. Eur. J. Immunol. 29, 4081–4091 (1999).

  12. 12

    Jensen, M., Tan, G., Forman, S., Wu, A. M. & Raubitschek, A. CD20 is a molecular target for scFvFc:zeta receptor redirected T cells: implications for cellular immunotherapy of CD20+ malignancy. Biol. Blood Marrow Transplant. 4, 75–83 (1998).

  13. 13

    Dembic, Z. et al. Transfer of specificity by murine alpha and beta T-cell receptor genes. Nature 320, 232–238 (1986).

  14. 14

    Clay, T. M. et al. Efficient transfer of a tumor antigen-reactive TCR to human peripheral blood lymphocytes confers anti-tumor reactivity. J. Immunol. 163, 507–513 (1999).

  15. 15

    Stanislawski, T. et al. Circumventing tolerance to a human MDM2-derived tumor antigen by TCR gene transfer. Nature Immunol. 2, 962–970 (2001).

  16. 16

    Orentas, R. J., Roskopf, S. J., Nolan, G. P. & Nishimura, M. I. Retroviral transduction of a T cell receptor specific for an Epstein–Barr virus-encoded peptide. Clin. Immunol. 98, 220–228 (2001).

  17. 17

    Fujio, K. et al. Functional reconstitution of class II MHC-restricted T cell immunity mediated by retroviral transfer of the alpha beta TCR complex. J. Immunol. 165, 528–532 (2000).

  18. 18

    Yang, L., Qin, X. F., Baltimore, D. & Van Parijs, L. Generation of functional antigen-specific T cells in defined genetic backgrounds by retrovirus-mediated expression of TCR cDNAs in hematopoietic precursor cells. Proc. Natl Acad. Sci. USA 99, 6204–6209 (2002).

  19. 19

    Kessels, H. W., Wolkers, M. C., van den Boom, M. D., van der Valk, M. A. & Schumacher, T. N. Immunotherapy through TCR gene transfer. Nature Immunol. 2, 957–961 (2001). In vivo tumour eradication by TCR-transduced mouse T cells, using a viral antigen-specific TCR.

  20. 20

    Chung, S., Wucherpfennig, K. W., Friedman, S. M., Hafler, D. A. & Strominger, J. L. Functional three-domain single-chain T-cell receptors. Proc. Natl Acad. Sci. USA 91, 12654–12658 (1994).

  21. 21

    Willemsen, R. A. et al. Grafting primary human T lymphocytes with cancer-specific chimeric single chain and two chain TCR. Gene Ther. 7, 1369–1377 (2000).

  22. 22

    Irving, B. A. & Weiss, A. The cytoplasmic domain of the T cell receptor zeta chain is sufficient to couple to receptor-associated signal transduction pathways. Cell 64, 891–901 (1991). Reports that the cytoplasmic domain of the CD3ζ chain can initiate T-cell activation in leukaemic T cells.

  23. 23

    Romeo, C. & Seed, B. Cellular immunity to HIV activated by CD4 fused to T cell or Fc receptor polypeptides. Cell 64, 1037–1046 (1991).

  24. 24

    Altenschmidt, U. et al. Cytolysis of tumor cells expressing the Neu/erbB-2, erbB-3, and erbB-4 receptors by genetically targeted naive T lymphocytes. Clin. Cancer Res. 2, 1001–1008 (1996).

  25. 25

    Muniappan, A., Banapour, B., Lebkowski, J. & Talib, S. Ligand-mediated cytolysis of tumor cells: use of heregulin-zeta chimeras to redirect cytotoxic T lymphocytes. Cancer Gene Ther. 7, 128–134 (2000).

  26. 26

    Eshhar, Z., Waks, T., Gross, G. & Schindler, D. G. Specific activation and targeting of cytotoxic lymphocytes through chimeric single chains consisting of antibody-binding domains and the gamma or zeta subunits of the immunoglobulin and T-cell receptors. Proc. Natl Acad. Sci. USA 90, 720–724 (1993). Shows that the antigen specificity of a CTL hybridoma can be redirected towards a hapten by scFV-based chimeric antigen receptors.

  27. 27

    Stancovski, I. et al. Targeting of T lymphocytes to Neu/HER2-expressing cells using chimeric single chain Fv receptors. J. Immunol. 151, 6577–6582 (1993).

  28. 28

    Geiger, T. L. & Jyothi, M. D. Development and application of receptor-modified T lymphocytes for adoptive immunotherapy. Transfus. Med. Rev. 15, 21–34 (2001).

  29. 29

    Ma, Q., Gonzalo–Daganzo, R. M., & Junghans, R. in Cancer Chemotherapy and Biological Response Modifiers, Annual 20 (eds Giaccone G, S. R. & Sondel P) 762 (Elsevier Science, New York, 2002).

  30. 30

    Altenschmidt, U., Klundt, E. & Groner, B. Adoptive transfer of in vitro-targeted, activated T lymphocytes results in total tumor regression. J. Immunol. 159, 5509–5515 (1997).

  31. 31

    Darcy, P. K. et al. Redirected perforin-dependent lysis of colon carcinoma by ex vivo genetically engineered CTL. J. Immunol. 164, 3705–3712 (2000).

  32. 32

    Haynes, N. M. et al. Single-chain antigen recognition receptors that costimulate potent rejection of established experimental tumors. Blood 100, 3155–3163 (2002).

  33. 33

    McGuinness, R. P. et al. Anti-tumor activity of human T cells expressing the CC49-zeta chimeric immune receptor. Hum. Gene Ther. 10, 165–173 (1999).

  34. 34

    Hwu, P. et al. In vivo antitumor activity of T cells redirected with chimeric antibody/T-cell receptor genes. Cancer Res. 55, 3369–3373 (1995).

  35. 35

    Wang, G. et al. A T cell-independent antitumor response in mice with bone marrow cells retrovirally transduced with an antibody/Fc-gamma chain chimeric receptor gene recognizing a human ovarian cancer antigen. Nature Med. 4, 168–172 (1998).

  36. 36

    Brentjens, R., Latouche, J. B., Riviere, I. & Sadelain, M. In vivo anti-tumor activity of genetically modified T cells is dependent on the method of ex vivo T cell expansion. Blood 100, 577a (2002). Demonstration of in vivo tumour eradication by systemic delivery of genetically targeted human PBLs. Also the first report to show that expansion in the presence of IL-15 is crucial for the therapeutic efficacy and persistence of adoptively transferred T cells.

  37. 37

    Patel, S. D. et al. Impact of chimeric immune receptor extracellular protein domains on T cell function. Gene Ther. 6, 412–419 (1999).

  38. 38

    Hombach, A. et al. Tumor-specific T cell activation by recombinant immunoreceptors: CD3 zeta signaling and CD28 costimulation are simultaneously required for efficient IL-2 secretion and can be integrated into one combined CD28/CD3 zeta signaling receptor molecule. J. Immunol. 167, 6123–6131 (2001).

  39. 39

    Mezzanzanica, D. et al. Transfer of chimeric receptor gene made of variable regions of tumor-specific antibody confers anticarbohydrate specificity on T cells. Cancer Gene Ther. 5, 401–407 (1998).

  40. 40

    Krause, A. et al. Antigen-dependent CD28 signaling selectively enhances survival and proliferation in genetically modified activated human primary T lymphocytes. J. Exp. Med. 188, 619–626 (1998). Reports that primary T-cell proliferation can be restored by a tumour-antigen-specific co-stimulatory receptor.

  41. 41

    Rossig, C., Bollard, C. M., Nuchtern, J. G., Rooney, C. M. & Brenner, M. K. Epstein–Barr virus-specific human T lymphocytes expressing antitumor chimeric T-cell receptors: potential for improved immunotherapy. Blood 99, 2009–2016 (2002).

  42. 42

    Ren-Heidenreich, L., Mordini, R., Hayman, G. T., Siebenlist, R. & LeFever, A. Comparison of the TCR zeta-chain with the FcR gamma-chain in chimeric TCR constructs for T cell activation and apoptosis. Cancer Immunol. Immunother. 51, 417–423 (2002).

  43. 43

    Moritz, D. & Groner, B. A spacer region between the single chain antibody- and the CD3 zeta-chain domain of chimeric T cell receptor components is required for efficient ligand binding and signaling activity. Gene Ther. 2, 539–546 (1995).

  44. 44

    Nolan, K. F. et al. Bypassing immunization: optimized design of 'designer T cells' against carcinoembryonic antigen (CEA)-expressing tumors, and lack of suppression by soluble CEA. Clin. Cancer Res. 5, 3928–3941 (1999).

  45. 45

    Geiger, T. L., Leitenberg, D. & Flavell, R. A. The TCR zeta-chain immunoreceptor tyrosine-based activation motifs are sufficient for the activation and differentiation of primary T lymphocytes. J. Immunol. 162, 5931–5939 (1999).

  46. 46

    Haynes, N. M. et al. Redirecting mouse CTL against colon carcinoma: superior signaling efficacy of single-chain variable domain chimeras containing TCR-zeta vs Fc epsilon RI-gamma. J. Immunol. 166, 182–187 (2001). Shows in vivo tumour eradication by systemic delivery of genetically targeted mouse T cells.

  47. 47

    Beecham, E. J., Ortiz-Pujols, S. & Junghans, R. P. Dynamics of tumor cell killing by human T lymphocytes armed with an anti-carcinoembryonic antigen chimeric immunoglobulin T-cell receptor. J. Immunother. 23, 332–343 (2000).

  48. 48

    Hombach, A. et al. T cells engrafted with a recombinant anti-CD30 receptor target autologous CD30(+) cutaneous lymphoma cells. Gene Ther. 8, 891–895 (2001).

  49. 49

    Yamada, G. et al. Retroviral expression of the human IL-2 gene in a murine T cell line results in cell growth autonomy and tumorigenicity. EMBO J. 6, 2705–2709 (1987).

  50. 50

    Karasuyama, H., Tohyama, N. & Tada, T. Autocrine growth and tumorigenicity of interleukin-2-dependent helper T cells transfected with IL-2 gene. J. Exp. Med. 1, 13–25 (1989).

  51. 51

    Treisman, J. et al. Interleukin-2-transduced lymphocytes grow in an autocrine fashion and remain responsive to antigen. Blood 85, 139–145 (1995).

  52. 52

    Hwu, P. & Rosenberg, S. A. The use of gene-modified tumor-infiltrating lymphocytes for cancer therapy. Ann. NY Acad. Sci. 716, 188–197 (1994).

  53. 53

    Hwu, P. et al. Functional and molecular characterization of tumor infiltrating lymphocytes transduced with tumor necrosis factor-α cDNA for the gene therapy of cancer in humans. J. Immunol. 150, 4104–4115 (1993).

  54. 54

    Liebowitz, D. N., Lee, K. P. & June, C. H. Costimulatory approaches to adoptive immunotherapy. Curr. Opin. Oncol. 10, 533–541 (1998).

  55. 55

    McAdam, A. J., Schweitzer, A. N. & Sharpe, A. H. The role of B7 co-stimulation in activation and differentiation of CD4+ and CD8+ T cells. Immunol. Rev. 165, 231–247 (1998).

  56. 56

    Chambers, C. A., Kuhns, M. S., Egen, J. G. & Allison, J. P. CTLA-4-mediated inhibition in regulation of T cell responses: mechanisms and manipulation in tumor immunotherapy. Annu. Rev. Immunol. 19, 565–594 (2001).

  57. 57

    Jenkins, M. K., Taylor, P. S., Norton, S. D. & Urdahl, K. B. CD28 delivers a costimulatory signal involved in antigen-specific IL-2 production by human T cells. J. Immunol. 147, 2461–2466 (1991).

  58. 58

    Staveley-O'Carroll, K. et al. Induction of antigen-specific T cell anergy: an early event in the course of tumor progression. Proc. Natl Acad. Sci. USA 95, 1178–1183 (1998).

  59. 59

    Croft, M., Bradley, L. M. & Swain, S. L. Naive versus memory CD4 T cell response to antigen. Memory cells are less dependent on accessory cell costimulation and can respond to many antigen-presenting cell types including resting B cells. J. Immunol. 152, 2675–2685 (1994).

  60. 60

    Scholz, C., Patton, K. T., Anderson, D. E., Freeman, G. J. & Hafler, D. A. Expansion of autoreactive T cells in multiple sclerosis is independent of exogenous B7 costimulation. J. Immunol. 160, 1532–1538 (1998).

  61. 61

    London, C. A., Lodge, M. P. & Abbas, A. K. Functional responses and costimulator dependence of memory CD4+ T cells. J. Immunol. 164, 265–272 (2000).

  62. 62

    Maric, M., Zheng, P., Sarma, S., Guo, Y. & Liu, Y. Maturation of cytotoxic T lymphocytes against a B7-transfected nonmetastatic tumor: a critical role for costimulation by B7 on both tumor and host antigen-presenting cells. Cancer Res. 58, 3376–3384 (1998).

  63. 63

    Ochsenbein, A. F. et al. Roles of tumour localization, second signals and cross priming in cytotoxic T-cell induction. Nature 411, 1058–1064 (2001).

  64. 64

    Gong, M. C. et al. Cancer patient T cells genetically targeted to prostate-specific membrane antigen specifically lyse prostate cancer cells and release cytokines in response to prostate-specific membrane antigen. Neoplasia 1, 123–127 (1999).

  65. 65

    Maher, J., Brentjens, R. J., Gunset, G., Riviere, I. & Sadelain, M. Human T-lymphocyte cytotoxicity and proliferation directed by a single chimeric TCRzeta /CD28 receptor. Nature Biotechnol. 20, 70–75 (2002).

  66. 66

    Hombach, A. et al. T-cell activation by recombinant receptors: CD28 costimulation is required for interleukin 2 secretion and receptor-mediated T-cell proliferation but does not affect receptor-mediated target cell lysis. Cancer Res. 61, 1976–1982 (2001). References 65, 66 and 69 show that cytotoxicity and proliferation can be redirected to tumour antigens by dual-signalling receptors that are derived from CD3ζ and CD28 in mouse and human primary T cells.

  67. 67

    Alvarez-Vallina, L. & Hawkins, R. E. Antigen-specific targeting of CD28-mediated T cell co-stimulation using chimeric single-chain antibody variable fragment-CD28 receptors. Eur. J. Immunol. 26, 2304–2309 (1996).

  68. 68

    Finney, H. M., Lawson, A. D., Bebbington, C. R. & Weir, A. N. Chimeric receptors providing both primary and costimulatory signaling in T cells from a single gene product. J. Immunol. 161, 2791–2797 (1998).

  69. 69

    Geiger, T. L., Nguyen, P., Leitenberg, D. & Flavell, R. A. Integrated src kinase and costimulatory activity enhances signal transduction through single-chain chimeric receptors in T lymphocytes. Blood 98, 2364–2371 (2001).

  70. 70

    Letterio, J. J. & Roberts, A. B. Regulation of immune responses by TGF-β. Annu. Rev. Immunol. 16, 137–161 (1998).

  71. 71

    Agrawal, B., Krantz, M. J., Reddish, M. A. & Longenecker, B. M. Cancer-associated MUC1 mucin inhibits human T-cell proliferation, which is reversible by IL-2. Nature Med. 4, 43–49 (1998).

  72. 72

    Harris, S. G. et al. Prostaglandins as modulators of immunity. Trends Immunol. 23, 144–150 (2002).

  73. 73

    Dong, H. et al. Tumor-associated B7-H1 promotes T cell apoptosis: a potential mechanism of immune evasion. Nature Med. 8, 793–800 (2002).

  74. 74

    Pasche, B. Role of TGF-β in cancer. J. Cell Physiol. 186, 153–168 (2001).

  75. 75

    Shariat, S. F. et al. Preoperative plasma levels of transforming growth factor-beta1 (TGF-β1) strongly predicts progression in patients undergoing radical prostatectomy. J. Clin. Oncol. 19, 2856–2864 (2001).

  76. 76

    Maeda, H. & Shiraishi, A. TGF-β contributes to the shift toward Th2-type responses through direct and IL10-mediated pathways in tumor-bearing mice. J. Immunol. 156, 73–78 (1996).

  77. 77

    Gorelik, L. & Flavell, R. A. Immune-mediated eradication of tumors through the blockade of TGF-β signaling in T cells. Nature Med. 7, 1118–1122 (2001). Shows that TGF-β blockade can improve CD8+ T-cell-mediated tumour elimination in transgenic mice.

  78. 78

    Bollard, C. M. et al. Adapting a TGF-β-related tumor protection strategy to enhance anti-tumor immunity. Blood 99, 3179–3187 (2002).

  79. 79

    Wieser, R., Attisano, L., Wrana, J. L. & Massague, J. Signaling activity of TGF-β type II receptors lacking specific domains in the cytoplasmic region. Mol. Cell. Biol. 15, 1573–1581 (1995).

  80. 80

    Cohen, J. L., Boyer, O., Thomas-Vaslin, V. & Klatzmann, D. Suicide gene-mediated modulation of graft-versus-host disease. Leuk. Lymph. 34, 473–480 (1999).

  81. 81

    Tiberghien, P. et al. Administration of herpes simplex-thymidine kinase-expressing donor T cells with a T-cell-depleted allogeneic marrow graft. Blood 97, 63–72 (2001).

  82. 82

    Marktel, S. et al. Immunologic potential of donor lymphocytes expressing a suicide gene for early immune reconstitution after hematopoietic T cell-depleted stem cell transplantation. Blood 3, 3 (2002).

  83. 83

    Sadelain, M. & Riviere, I. Sturm und drang over suicidal lymphocytes. Mol. Ther. 5, 655–657 (2002).

  84. 84

    Gallardo, H. F., Tan, C. & Sadelain, M. The internal ribosomal entry site of the encephalomyocarditis virus enables reliable coexpression of two transgenes in human primary T lymphocytes. Gene Ther. 4, 1115–1119 (1997).

  85. 85

    Sadelain, M. & Blasberg, R. G. Imaging transgene expression for gene therapy. J. Clin. Pharmacol. 39, 34S–39S (1999).

  86. 86

    Tjuvajev, J. G., et al. Imaging hsvtk gene transfer and expression by positron emission tomography. Cancer Res. 58, 4333–4341 (1995).

  87. 87

    Koehne, G. et al. In vivo imaging of the targeted migration of hsvtk transduced human antigen-specific T lymphocytes. Nature Biotechnol. (in the press).

  88. 88

    Hardy, J. et al. Bioluminescence imaging of lymphocytes trafficking in vivo. Exp. Hematol. 29, 1353–1360 (2001).

  89. 89

    Ponomarev, V. et al. Imaging TCR-dependent NFAT-mediated T-cell activation with positron emission tomography in vivo. Neoplasia 3, 480–488 (2001). Demonstration of non-invasive imaging of T-cell activation in leukaemic T cells in vivo.

  90. 90

    Gambhir, S. S. et al. Imaging transgene expression with radionuclide imaging technologies. Neoplasia 2, 118–138 (2000).

  91. 91

    Bogdanov, A. & Weissleder, R. In vivo imaging of gene delivery and expression. Trends Biotechnol. 20, S11–S18 (2002).

  92. 92

    Nanda, N. K. & Sercarz, E. E. Induction of anti-self-immunity to cure cancer. Cell 82, 13–17 (1995).

  93. 93

    Speiser, D. E. et al. Self antigens expressed by solid tumors do not efficiently stimulate naive or activated T cells: implications for immunotherapy. J. Exp. Med. 186, 645–653 (1997).

  94. 94

    Ochsenbein, A. F. et al. Immune surveillance against a solid tumor fails because of immunological ignorance. Proc. Natl Acad. Sci. USA 96, 2233–2238 (1999).

  95. 95

    Vasmel, W. L. E. et al. Primary virus-induced lymphomas evade T cell immunity by failure to express viral antigens. J. Exp. Med. 169, 1233–1254 (1989).

  96. 96

    Dunn, G. P., Bruce, A. T., Ikeda, H., Old, L. J. & Schreiber, R. D. Cancer immunoediting: from immunosurveillance to tumor escape. Nature Immunol. 3, 991–998 (2002).

  97. 97

    Garrido, F. et al. Implications for immunosurveillance of altered HLA class I phenotypes in human tumours. Immunol. Today 18, 89–95 (1997).

  98. 98

    Motyka, B. et al. Mannose 6-phosphate/IGF II receptor is a death receptor for granzyme B during cytotoxic T cell-induced apoptosis. Cell 103, 491–500 (2000).

  99. 99

    Medema, J. P. et al. Blockade of the granzyme B/perforin pathway through overexpression of the serine protease inhibitor PI-9/SPI-6 constitutes a mechanism for immune escape by tumors. Proc. Natl Acad. Sci. USA 98, 11515–11520 (2001).

  100. 100

    Correa, M. R. et al. Sequential development of structural and functional alterations in T cells from tumor-bearing mice. J. Immunol. 158, 5292–5296 (1997).

  101. 101

    Mitsuyatsu, R. T. et al. Prolonged survival and tissue trafficking following adoptive therapy of CD4z gene-modified autologous CD4+ and CD8+ T cells in HIV-infected subjects. Blood 96, 785–793 (2000).

  102. 102

    Dudley, M. E. et al. Cancer regression and autoimmunity in patients after clonal repopulation with anti-tumor lymphocytes. Science 298, 850–854 (2002).

  103. 103

    Bretscher, P. & Cohn, M. A theory of self-nonself discrimination. Science 169, 1042–1049 (1970).

  104. 104

    Jenkins, M. K. & Schwartz, R. H. Antigen presentation by chemically modified splenocytes induces antigen-specific T cell unresponsiveness in vitro and in vivo. J. Exp. Med. 165, 302–319 (1987).

  105. 105

    Salomon, B. & Bluestone, J. A. Complexities of CD28/B7:CTLA-4 costimulatory pathways in autoimmunity and transplantation. Annu. Rev. Immunol. 19, 225–252 (2001).

  106. 106

    Lindsten, T., June, C. H., Ledbetter, J. A., Stella, G. & Thompson, C. B. Regulation of lymphokine messenger RNA stability by a surface-mediated T cell activation pathway. Science 244, 339–343 (1989).

  107. 107

    Fraser, J. D., Irving, B. A., Crabtree, G. R. & Weiss, A. Regulation of interleukin-2 gene enhancer activity by the T cell accessory molecule CD28. Science 251, 313–316 (1991).

  108. 108

    Boise, L. H. et al. CD28 costimulation can promote T cell survival by enhancing the expression of Bcl-XL . Immunity 3, 87–98 (1995).

  109. 109

    Kearney, E. R. et al. Antigen-dependent clonal expansion of a trace population of antigen-specific CD4+ T cells in vivo is dependent on CD28 costimulation and inhibited by CTLA-4. J. Immunol. 155, 1032–1036 (1995).

  110. 110

    Viola, A. & Lanzavecchia, A. T cell activation determined by T cell receptor number and tunable thresholds. Science 273, 104–106 (1996).

  111. 111

    Wulfing, C. & Davis, M. M. A receptor/cytoskeletal movement triggered by costimulation during T cell activation. Science 282, 2266–2269 (1998).

  112. 112

    Viola, A., Schroeder, S., Sakakibara, Y. & Lanzavecchia, A. T lymphocyte costimulation mediated by reorganization of membrane microdomains. Science 283, 680–682 (1999).

  113. 113

    Pinkoski, M. J. & Green, D. R. Lymphocyte apoptosis: refining the paths to perdition. Curr. Opin. Hematol. 9, 43–49 (2002).

  114. 114

    Boussiotis, V. A., Freeman, G. J., Gray, G., Gribben, J. & Nadler, L. M. B7 but not intercellular adhesion molecule-1 costimulation prevents the induction of human alloantigen-specific tolerance. J. Exp. Med. 178, 1753–1763 (1993).

  115. 115

    Germain, R. N. & Stefanova, I. The dynamics of T cell receptor signaling: complex orchestration and the key roles of tempo and cooperation. Annu. Rev. Immunol. 17, 467–522 (1999).

  116. 116

    Heath, W. R. & Carbone, F. R. Cross-presentation, dendritic cells, tolerance and immunity. Annu. Rev. Immunol. 19, 47–64 (2001).

  117. 117

    Townsend, S. E. & Allison, J. P. Tumor rejection after direct costimulation of CD8+ T cells by B7-transfected melanoma cells. Science 259, 368–370 (1993).

  118. 118

    Mandelbrot, D. A., Kishimoto, K., Auchincloss, H., Sharpe, A. H. & Sayegh, M. H. Rejection of mouse cardiac allografts by costimulation in trans. J. Immunol. 167, 1174–1178 (2001).

  119. 119

    Beecham, E. J., Ma, Q., Ripley, R. & Junghans, R. P. Coupling CD28 co-stimulation to immunoglobulin T-cell receptor molecules: the dynamics of T-cell proliferation and death. J Immunother. 23, 631–642 (2000).

  120. 120

    Hombach, A. et al. An entirely humanized CD3 zeta-chain signaling receptor that directs peripheral blood T cells to specific lysis of carcinoembryonic antigen-positive tumor cells. Int. J. Cancer 88, 115–120 (2000).

  121. 121

    Daly, T. et al. Recognition of human colon cancer by T cells transduced with a chimeric receptor gene. Cancer Gene Ther. 7, 284–291 (2000).

  122. 122

    Patel, S. D., Ge, Y., Moskalenko, M. & McArthur, J. G. Anti-tumor CC49-zeta CD4 T cells possess both cytolytic and helper functions. J. Immunother. 23, 661–668 (2000).

  123. 123

    Yun, C. O., Nolan, K. F., Beecham, E. J., Reisfeld, R. A. & Junghans, R. P. Targeting of T lymphocytes to melanoma cells through chimeric anti-GD3 immunoglobulin T-cell receptors. Neoplasia 2, 449–459 (2000).

  124. 124

    Parker, L. L. et al. Expansion and characterization of T cells transduced with a chimeric receptor against ovarian cancer. Hum. Gene Ther. 11, 2377–2387 (2000).

  125. 125

    Weijtens, M. E., Willemsen, R. A., Valerio, D., Stam, K. & Bolhuis, R. L. Single chain Ig/gamma gene-redirected human T lymphocytes produce cytokines, specifically lyse tumor cells, and recycle lytic capacity. J. Immunol. 157, 836–843 (1996).

  126. 126

    Weijtens, M. E., Hart, E. H. & Bolhuis, R. L. Functional balance between T cell chimeric receptor density and tumor associated antigen density: CTL mediated cytolysis and lymphokine production. Gene Ther. 7, 35–42 (2000).

  127. 127

    Weijtens, M. E., Willemsen, R. A., van Krimpen, B. A. & Bolhuis, R. L. Chimeric scFv/gamma receptor-mediated T-cell lysis of tumor cells is coregulated by adhesion and accessory molecules. Int. J. Cancer 77, 181–187 (1998).

  128. 128

    Ren-Heidenreich, L., Hayman, G. T. & Trevor, K. T. Specific targeting of EGP-2+ tumor cells by primary lymphocytes modified with chimeric T cell receptors. Hum. Gene Ther. 11, 9–19 (2000).

  129. 129

    Kershaw, M. H. et al. Generation of gene-modified T cells reactive against the angiogenic kinase insert domain-containing receptor (KDR) found on tumor vasculature. Hum. Gene Ther. 11, 2445–2452 (2000).

  130. 130

    Niederman, T. M. et al. Antitumor activity of cytotoxic T lymphocytes engineered to target vascular endothelial growth factor receptors. Proc. Natl Acad. Sci. USA 99, 7009–7014 (2002).

Download references


Our work is primarily supported by National Institutes of Health grants and the Goodwin ETC fund. We thank K. Hurdle and E. Ciccaroni for excellent assistance with the preparation of the manuscript and the figures.

Author information

Correspondence to Michel Sadelain.

Related links



(CTLs). T lymphocytes that exert a cytolytic function following engagement of their T-cell antigen receptor on target cells. CTLs express the co-receptor CD8 and recognize antigenic peptides (or CTL epitopes) that are presented by HLA class I molecules.


A state of unresponsiveness to antigen that can be induced when the T-cell receptor binds antigen in the absence of co-stimulation.


A cell line that is created by the fusion of a lymphocyte tumour cell to an antigen-specific T-cell clone. This fusion results in a T-cell line that maintains the specificity of the T-cell clone as well as the immortalized phenotype of the tumour cell.


Primary T cells are cells that have been freshly harvested from the donor and have not undergone transformation or extensive culture. Hybridomas, leukaemias and lymphomas are transformed cells that might not faithfully reflect or recapitulate the activation requirements or differentiation of primary T cells. T-cell lines are obtained by periodic restimulation of cultured primary T cells. T-cell clones are the progeny of a single primary T cell.


(HLAs). Cell-surface molecules that are encoded by the major histocompatibility complex. These molecules present antigenic peptides to T cells via their T-cell antigen receptor. HLA class I molecules present antigen to CD8+ T lymphocytes, and HLA class II molecules present antigen to CD4+ lymphocytes.


T lymphocytes that exert regulatory and helper functions for B cells, cytotoxic T lymphocytes and other immune effector cells. Helper T cells express the co-receptor CD4, which recognizes antigenic peptides (or helper epitopes) that are presented by HLA class II molecules.


Long-lived clonal T cells that develop following a primary immune response. This T-cell population mediates the secondary T-cell response that is seen following late re-exposure to the cognate antigen.


A term that is used to describe the supra-molecular structure that is established between a T cell and an antigen-presenting cell (APC) bearing the HLA-peptide complex recognized by the T-cell antigen receptor (TCR). At the centre of the synapse, adhesion is mediated through binding of the TCR to the APC's HLA-peptide complex. Peripherally, this complex is stabilized through additional interactions — for example, between LFA1 and ICAM1.

Rights and permissions

Reprints and Permissions

About this article

Further reading

Figure 1: Structure of physiological antigen receptors.
Figure 2: Examples of chimeric antigen receptors.
Figure 3: Co-stimulation of tumour-reactive T cells in vivo.