Abstract
The discovery of multiple costimulatory cell surface molecules that influence the course of T cell activation has increased our appreciation of the complexity of the T cell response. It remains clear, however, that CD28 and cytotoxic T lymphocyte antigen 4 (CTLA-4) are the critical costimulatory receptors that determine the early outcome of stimulation through the T cell antigen receptor (TCR). Details of how the T cell integrates TCR stimulation with the costimulatory signals of CD28 and the inhibitory signals of CTLA-4 remain to be established, but unique features of the cell biology of CTLA-4 provide important insights into its function. We summarize here recent findings that suggest a previously unrecognized role for CTLA-4 in the regulation of T cell responses. We also describe preclinical and clinical results that indicate manipulation of CTLA-4 has considerable promise as a strategy for the immunotherapy of cancer.
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References
Mueller, D.L., Jenkins, M.K. & Schwartz, R.H. Clonal expansion versus functional clonal inactivation: A costimulatory signalling pathway determines the outcome of T cell antigen receptor occupancy. Annu. Rev. Immunol. 7, 445–480 (1989).
Sharpe, A.H. & Freeman, G.J. The B7-CD28 superfamily. Nature Immunol. 2, 116–126 (2002).
Brunet, J.F. et al. A new member of the immunoglobulin superfamily–CTLA-4. Nature 328, 267–270 (1987).
Linsley, P.S. et al. CTLA-4 is a second receptor for the B cell activation antigen B7. J. Exp. Med. 174, 561–569 (1991).
Linsley, P.S. et al. Human B7-1 (CD80) and B7-2 (CD86) bind with similar avidities but distinct kinetics to CD28 and CTLA-4 receptors. Immunity 1, 793–801 (1994).
van der Merwe, P.A., Bodian, D.L., Daenke, S., Linsley, P. & Davis, S.J. CD80 (B7-1) binds both CD28 and CTLA-4 with a low affinity and very fast kinetics. J. Exp. Med. 185, 393–403 (1997).
Linsley, P.S. et al. Coexpression and functional cooperativity of CTLA-4 and CD28 on activated T lymphocytes. J. Exp. Med. 176, 1595–1604 (1992).
Walunas, T.L. et al. CTLA-4 can function as a negative regulator of T cell activation. Immunity 1, 405–413 (1994).
Krummel, M.F. & Allison, J.P. CD28 and CTLA-4 have opposing effects on the response of T cells to stimulation. J. Exp. Med. 182, 459–465 (1995).
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, 1033–1036 (1995).
Krummel, M.F., Sullivan, T.J. & Allison, J.P. Superantigen responses and costimulation: CD28 and CTLA-4 have opposing effects on T cell expansion in vitro and in vivo. Int. Immunol. 8, 519–523 (1996).
Waterhouse, P. et al. Lymphoproliferative disorders with early lethality in mice deficient in CTLA-4. Science 270, 985–988 (1995).
Tivol, E.A. et al. Loss of CTLA-4 leads to massive lymphoproliferation and fatal multiorgan tissue destruction, revealing a critical negative regulatory role of CTLA-4. Immunity 3, 541–547 (1995).
Chambers, C.A., Sullivan, T.J. & Allison, J.P. Lymphoproliferation in CTLA-4-deficient mice is mediated by costimulation-dependent activation of CD4+ T cells. Immunity 7, 885–895 (1997).
Lindsten, T. et al. Characterization of CTLA-4 structure and expression on human T cells. J. Immunol. 151, 3489–3499 (1993).
Brunner, M.C. et al. CTLA-4 mediated inhibition of early events of T cell proliferation. J. Immunol. 162, 5813–5820 (1999).
Iezzi, G., Karjalainen, K. & Lanzavecchia, A. The duration of antigenic stimulation determines the fate of naive and effector T cells. Immunity 8, 89–95 (1998).
Lee, K.H. et al. T cell receptor signaling precedes immunological synapse formation. Science 295, 1539–1542 (2002).
Egen, J.G. & Allison, J.P. Cytotoxic T lymphocyte antigen-4 accumulation in the immunological synapse is regulated by TCR signal strength. Immunity 16, 23–35 (2002).
Schneider, H. et al. Cytolytic T lymphocyte-associated antigen-4 and the TCRζ/CD3 complexes, but not CD28, interact with clathrin adaptor complexes AP-1 and AP-2. J. Immunol. 163, 1868–1879 (1999).
Oki, S., Kohsaka, T. & Azuma, M. Augmentation of CTLA-4 expression by wortmannin: involvement of lysosomal sorting properties of CTLA-4. Int. Immunol. 11, 1563–1571 (1999).
Iida, T. et al. Regulation of cell surface expression of CTLA-4 by secretion of CTLA-4- containing lysosomes upon activation of CD4+ T cells. J. Immunol. 165, 5062–5068 (2000).
Blott, E.J. & Griffiths, G.M. Secretory lysosomes. Nature Rev. Mol. Cell Biol. 3, 122–31 (2002).
Truitt, K.E., Hicks, C.M. & Imboden, J.B. Stimulation of CD28 triggers an association between CD28 and phospatidylinositol 3-kinase in Jurkat T cells. J. Exp. Med. 179, 1071–1076 (1994).
Prasad, K.V. et al. T-cell antigen CD28 interacts with the lipid kinase phosphatidylinositol 3-kinase by a cytoplasmic Tyr(P)-Met-Xaa-Met motif. Proc. Natl. Acad. Sci. USA 91, 2834–2838 (1994).
Schneider, H., Prasad, V.S., Shoelson, S.E. & Rudd, C.E. CTLA-4 binding to the lipid kinase phosphatidylinositol 3-kinase in T cells. J. Exp. Med. 181, 351–355 (1995).
Leung, H.T., Bradshaw, J., Cleaveland, J.S. & Linsley, P.S. Cytotoxic T lymphocyte-associated molecule-4, a high avidity receptor for CD80 and CD86, contains an intracellular localization motif in its cytoplasmic tail. J. Biol. Chem. 270, 25107–25114 (1995).
Shiratori, T. et al. Tyrosine phosphorylation controls internalization of CTLA-4 by regulating its interaction with clathrin-associated adaptor complex AP-2. Immunity 6, 583–589 (1997).
Chuang, E. et al. Interaction of CTLA-4 with the clathrin-associated protein AP50 results in ligand-independent endocytosis that limits cell surface expression. J. Immunol. 159, 144–151 (1997).
Zhang, Y. & Allison, J.P. Interaction of CTLA-4 with AP50, a clathrin-coated pit adaptor protein. Proc. Natl. Acad. Sci. USA 94, 9273–9278 (1997).
Bradshaw, J.D. et al. Interaction of the cytoplasmic tail of CTLA-4 (CD152) with a clathrin- associated protein is negatively regulated by tyrosine phosphorylation. Biochemistry 36, 15975–15982 (1997).
Kupfer, A., Swain, S.L. & Singer, S.J. The specific direct interaction of helper T cells and antigen-presenting B cells. II. Reorientation of the microtubule organizing center and reorganization of the membrane-associated cytoskeleton inside the bound helper T cells. J. Exp. Med. 165, 1565–1580 (1987).
Linsley, P.S. et al. Intracellular trafficking of CTLA-4 and focal localization towards sites of TCR engagement. Immunity 4, 535–543 (1996).
Alegre, M.L., Frauwirth, K.A. & Thompson, C.B. T-cell regulation by CD28 and CTLA-4. Nature Rev. Immunol. 1, 220–228 (2001).
Schwartz, J.-C.D., Zhang, X., Nathenson, S.G. & Almo, S.C. Structural mechanisms of costimulation. Nature Immunol. 2, 427–434 (2002).
Krummel, M.F. & Allison, J.P. CTLA-4 engagement inhibits IL-2 accumulation and cell cycle progression upon activation of resting T cells. J. Exp. Med. 183, 2533–2540 (1996).
Walunas, T.L., Bakker, C.Y. & Bluestone, J.A. CTLA-4 ligation blocks CD28-dependent T cell activation. J. Exp. Med. 183, 2541–2550 (1996).
Greenwald, R.J. et al. CTLA-4 regulates cell cycle progression during a primary immune response. Eur. J. Immunol. 32, 366–373 (2002).
Bachmann, M.F., Köhler, G., Ecabert, B., Mak, T.W. & Kopf, M. Cutting edge: lymphoproliferative disease in the absence of CTLA-4 is not T cell autonomous. J. Immunol. 163, 1128–1131 (1999).
Bachmann, M.F. et al. Normal pathogen-specific immune responses mounted by CTLA-4-deficient T cells: a paradigm reconsidered. Eur. J. Immunol. 31, 450–458 (2001).
Salomon, B. et al. B7/CD28 costimulation is essential for the homeostasis of the CD4+CD25+ immunoregulatory T cells that control autoimmune diabetes. Immunity 12, 431–440 (2000).
Read, S., Malmstrom, V. & Powrie, F. Cytotoxic T lymphocyte-associated antigen 4 plays an essential role in the function of CD25+CD4+ regulatory cells that control intestinal inflammation. J. Exp. Med. 192, 295–302 (2000).
Takahashi, T. et al. Immunologic self-tolerance maintained by CD25+CD4+ regulatory T cells constitutively expressing cytotoxic T lymphocyte-associated antigen 4. J. Exp. Med. 192, 303–310 (2000).
Sutmuller, R.P.M. et al. Synergism of CTLA-4 blockade and depletion of CD25+ regulatory T cells in anti-tumor therapy reveals alternative pathways for suppression of auto-reactive CTL responses. J. Exp. Med. 194, 823–832 (2001).
Greenwald, R.J., Boussiotis, V.A., Lorsbach, R.B., Abbas, A.K. & Sharpe, A. CTLA-4 regulates induction of anergy in vivo. Immunity 14, 145–155 (2001).
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).
Chambers, C.A. et al. The role of CTLA-4 in the regulation and initiation of T cell responses. Immunol. Rev. 153, 27–46 (1996).
Fallarino, F., Fields, P.E. & Gajewski, T.F. B7-1 engagement of cytotoxic T lymphocyte antigen 4 inhibits T cell activation in the absence of CD28. J. Exp. Med. 188, 205–210 (1998).
Gajewski, T.F., Fallarino, F., Fields, P.E., Rivas, F. & Alegre, M.L. Absence of CTLA-4 lowers the activation threshold of primed CD8+ TCR-transgenic T cells: lack of correlation with Src homology domain 2-containing protein tyrosine phosphatase. J. Immunol. 166, 3900–3907 (2001).
Chambers, C.A., Kuhns, M.S. & Allison, J.P. Cytotoxic T lymphocyte antigen-4 (CTLA-4) regulates primary and secondary peptide-specific CD4+ T cell responses. Proc. Natl. Acad. Sci. USA 96, 8603–8608 (1999).
Tivol, E.A. et al. CTLA4Ig prevents lymphoproliferation and fatal multiorgan tissue destruction in CTLA-4-deficient mice. J. Immunol. 158, 5091–5094 (1997).
Mandelbrot, D.A., McAdam, A.J. & Sharpe, A.H. B7-1 or B7-2 is required to produce the lymphoproliferative phenotype in mice lacking cytotoxic T lymphocyte-associated antigen 4 (CTLA-4). J. Exp. Med. 189, 435–440 (1999).
Masteller, E.L., Chuang, E., A.C., M., Reiner, S.L. & Thompson, C.B. Structural analysis of CTLA-4 function in vivo. J. Immunol. 164, 5319–5327 (2000).
Doyle, A.M. et al. Induction of cytotoxic T lymphocyte antigen 4 (CTLA-4) restricts clonal expansion of helper T cells. J. Exp. Med. 194, 893–902 (2001).
Salomon, B. & Bluestone, J.A. Complexities of CD28/B7: CTLA-4 costimulatory pathways in autoimmunity and transplantation. Annu. Rev. Immunol. 19, 225–252 (2001).
Perez, V.L. et al. Induction of peripheral T cell tolerance in vivo requires CTLA-4 engagement. Immunity 6, 411–417 (1997).
Murphy, M.L., Cotterell, S.E., Gorak, P.M., Engwerda, C.R. & Kaye, P.M. Blockade of CTLA-4 enhances host resistance to the intracellular pathogen, Leishmania donovani. J. Immunol. 161, 4153–4160 (1998).
McCoy, K., Camberis, M. & Gros, G.L. Protective immunity to nematode infection is induced by CTLA-4 blockade. J. Exp. Med. 186, 183–187 (1997).
Kuhns, M.S., Epshteyn, V., Sobel, R.A. & Allison, J.P. CTLA-4 regulates the size, function, and reactivity of a primed pool of T cells. Proc. Natl. Acad. Sci. USA 97, 12711–12716 (2000).
Busch, D.K. & Pamer, E.G. T cell affinity maturation by selective expansion during infection. J. Exp. Med. 189, 701–709 (1999).
Savage, P.A., Boniface, J.J. & Davis, M.M. A kinetic basis for T cell receptor repertoire selection during an immune response. Immunity 10, 485–492 (1999).
Butz, E.A. & Bevan, M.J. Massive expansion of antigen-specific CD8+ T cells during an acute virus infection. Immunity 8, 167–175 (1998).
Bousso, P., Levraud, J.P., Kourilsky, P. & Abastado, J.P. The composition of a primary T cell response is largely determined by the timing of recruitment of individual T cell clones. J. Exp. Med. 189, 1591–1600 (1999).
Kedl, R.M. et al. T cells compete for access to antigen-bearing antigen-presenting cells. J. Exp. Med. 192, 1105–1113 (2000).
Rogers, P.R. & Croft, M. Peptide dose, affinity, and time of differentiation can contribute to the Th1/Th2 cytokine balance. J. Immunol. 163, 1205–1213 (1999).
Malherbe, L. et al. Selective activation and expansion of high-affinity CD4+ T cells in resistant mice upon infection with Leishmania major. Immunity 13, 771–782 (2000).
Hurwitz, A.A., Sullivan, T.J., Sobel, R.A. & Allison, J.P. Cytotoxic T lymphocyte antigen-4 (CTLA-4) limits the expansion of encephalitogenic T cells in experimental autoimmune encephalomyelitis (EAE)-resistant BALB/c mice. Proc. Natl. Acad. Sci. USA 99, 3013–3017 (2002).
Vanasek, T.L., Khoruts, A., Zell, T. & Mueller, D.L. Antagonistic roles for CTLA-4 and the mammalian target of rapamycin in the regulation of clonal anergy: enhanced cell cycle progression promotes recall antigen responsiveness. J. Immunol. 167, 5636–5644 (2001).
Chen, L. et al. Costimulation of antitumor immunity by the B7 counterreceptor for the T lymphocyte molecules CD28 and CTLA-4. Cell 71, 1093–1102 (1992).
Townsend, S. & Allison, J.P. Tumor rejection after direct costimulation of CD8+ T cells by B7-transfected melanoma cells. Science 259, 368–370 (1993).
Huang, A.Y. et al. Role of bone marrow-derived cells in presenting MHC class I-restricted tumor antigens. Science 264, 961–965 (1994).
Leach, D., Krummel, M. & Allison, J.P. Enhancement of antitumor immunity by CTLA-4 blockade. Science 271, 1734–1736 (1996).
Kwon, E.D. et al. Manipulation of T cell costimulatory and inhibitory signals for immunotherapy of prostate cancer. Proc. Natl. Acad. Sci. USA 94, 8099–8103 (1997).
Yang, Y. et al. Enhanced induction of antitumor T-cell responses by cytotoxic T lymphocyte associated molecule-4 blockade: The effect is manifested only at the restricted tumor-bearing stages. Cancer Res. 57, 4036–4041 (1997).
Shrikant, P., Khoruts, A. & Mescher, M.F. CTLA-4 blockade reverses CD8+ T cell tolerance to tumor by a CD4+ T cell and IL-2 dependent mechanism. Immunity 11, 483–493 (1999).
Sotomayor, E.M., Borrello, I.M., Tubb, E., Allison, J.P. & Levitsky, H.I. In vivo blockade of CTLA-4 enhances the priming of responsive T-cells but fails to prevent the induction of tumor antigen-specific tolerance. Proc. Natl. Acad. Sci. USA 96, 11476–11481 (1999).
Hurwitz, A.A. et al. in Cytotoxic cells: basic mechanisms and medical applications (eds. Sitovsky, M. V. & Henkart, P. A.) 385–393 (Lippincott Williams & Wilkins, Philadelphia, PA, 1999).
Hurwitz, A.A., Yu, T.F., Leach, D.R. & Allison, J.P. CTLA-4 blockade synergizes with tumor-derived GM-CSF for treatment of an experimental mammary carcinoma. Proc. Natl. Acad. Sci. USA 95, 10067–10071 (1998).
van Elsas, A., Hurwitz, A.A. & Allison, J.P. Combination immunotherapy of B16 melanoma using anti–CTLA-4 and GM-CSF producing vaccines induces rejection of subcutaneous and metastatic tumors accompanied by autoimmune depigmentation. J. Exp. Med. 190, 355–366 (1999).
Dranoff, G. et al. Vaccination with irradiated tumor cells engineered to secrete GM-CSF stimulates potent, specific, and long lasting anti-tumor immunity. Proc. Natl. Acad. Sci. USA 90, 3539–3543 (1993).
Hurwitz, A.A. et al. Combination immunotherapy of primary prostate cancer in a transgenic model using CTLA-4 blockade. Cancer Res. 60, 2444–2448 (2000).
Rosenberg, S.A. & White, D.E. Vitiligo in patients with melanoma: normal tissue antigens can be targets for cancer immunotherapy. J. Immunother. EmphasisTumor Immunol. 19, 81–84 (1996).
van Elsas, A. et al. Elucidating the autoimmune and anti-tumor effector mechanisms of a treatment based on cytotoxic T lymphocyte antigen-4 (CTLA-4) blockade in combination with a B16 melanoma vaccine: Comparison of prophylaxis and therapy. J. Exp. Med. 194, 427–438 (2001).
Hung, K. et al. The central role of CD4+ T cells in the antitumor immune response. J. Exp. Med. 188, 2357–2368 (1998).
Karandikar, N.J., Vanderlugt, C.L., Walunas, T.L., Miller, S.D. & Bluestone, J.A. CTLA-4: A negative regulator of autoimmune disease. J. Exp. Med. 184, 783–788 (1996).
Perrin, P.J., Maldonado, J.H., Davis, T.A., June, C.H. & Racke, M.K. CTLA-4 blockade enhances clinical disease and cytokine production during experimental allergic encephalomyelitis. J. Immunol. 157, 1333–1336 (1996).
Hurwitz, A.A., Sullivan, T.J., Krummel, M.F., Sobel, R.A. & Allison, J.P. Specific blockade of CTLA-4/B7 interactions results in exacerbated clinical and histologic disease in an actively-induced model of experimental allergic encephalomyelitis. J. Neuroimmunol. 73, 57–62 (1997).
Lühder, F., Höglund, P., Allison, J.P., Benoist, C. & Mathis, D. Cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) regulates the unfolding of autoimmune diabetes. J. Exp. Med. 187, 427–432 (1998).
Mokyr, M.B., Kalinichenko, T., Gorelik, L. & Bluestone, J.A. Realization of the therapeutic potential of CTLA-4 blockade in low-dose chemotherapy-treated tumor-bearing mice. Cancer Res. 58, 5301–5304 (1998).
Mercader, M. et al. T cell infiltration of the prostate induced by androgen withdrawal in patients with prostate cancer. Proc. Natl. Acad. Sci. USA 98, 14565–14570 (2001).
Acknowledgements
We thank P. Savage and J. Engelhardt for helpful discussions and critical reading of the manuscript and T. Sullivan, M. Fasso, C. Chambers, A. Hurwitz, A. van Elsas and E. Kwon for helpful discussions. Supported by the Howard Hughes Medical Institute and grants from the NIH (NCI CA40041 and CA57986) and CaPCURE (J. P. A.).
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Egen, J., Kuhns, M. & Allison, J. CTLA-4: new insights into its biological function and use in tumor immunotherapy. Nat Immunol 3, 611–618 (2002). https://doi.org/10.1038/ni0702-611
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DOI: https://doi.org/10.1038/ni0702-611
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