The Journal of Immunology, Vol 151, Issue 7 3489-3499, Copyright © 1993 by American Association of Immunologists

ARTICLES

Characterization of CTLA-4 structure and expression on human T cells

T Lindsten, KP Lee, ES Harris, B Petryniak, N Craighead, PJ Reynolds, DB Lombard, GJ Freeman, LM Nadler and GS Gray
Department of Pathology, University of Michigan, Ann Arbor 48109.

CTLA-4 is an adhesion receptor expressed on activated T cells. The amino acid sequence of CTLA-4 is related to CD28, and although the function of CTLA-4 remains unknown, it shares several features with CD28, including a common counter-receptor, B7, that is present on Ag- presenting cells. In a recent study we found that CD28 and CTLA-4 were coexpressed at the mRNA level on activated T cells but that only CD28 was expressed on resting T cells. Here we show that within the T cell population, CTLA-4 expression is restricted to the subset of T cells that also express cell surface CD28. CTLA-4 mRNA expression can be induced on quiescent T cells via phorbol ester-mediated activation of protein kinase C but not with calcium ionophore treatment alone. Phorbol ester-induced expression of CTLA-4 mRNA could be enhanced with calcium ionophore treatment, and treatment of cells in this manner resulted in a reciprocal decrease in expression of CD28 mRNA. Ligation of CD28 with monoclonal antibody also resulted in the specific and rapid induction of CTLA-4 mRNA. To study the expression of CTLA-4 at the protein level, a rabbit antiserum against a recombinant protein derived from CTLA-4 cDNA was generated. When activated T cells were labeled with [35S]methionine, the rabbit antiserum precipitated a 41- to 43-kDa protein from whole cell lysates. Similar results were found when detergent-soluble lysates from 125I surface-labeled resting and activated T cells were analyzed by SDS-PAGE. Surprisingly, under the conditions tested, CTLA-4 migrated primarily as a monomer at the cell surface, and could not be shown to exist as a disulfide-bonded homodimer or as a heterodimer consisting of CTLA-4 and CD28. These results suggest that B7 can bind to T cells via distinct receptor complexes consisting of either CD28 or CTLA-4, and that these complexes may potentially mediate distinct biologic functions. Further, the present results suggest that noncovalent interactions might mediate association of CTLA-4 and/or CD28 at the cell surface.

This article has been cited by other articles:

				A. Huber, F. Menconi, S. Corathers, E. M. Jacobson, and Y. Tomer Joint Genetic Susceptibility to Type 1 Diabetes and Autoimmune Thyroiditis: from Epidemiology to Mechanisms Endocr. Rev., October 1, 2008; 29(6): 697 - 725. [Abstract] [Full Text] [PDF]

				M. Berg and N. Zavazava Regulation of CD28 expression on CD8+ T cells by CTLA-4 J. Leukoc. Biol., April 1, 2008; 83(4): 853 - 863. [Abstract] [Full Text] [PDF]

				S. G. Downey, J. A. Klapper, F. O. Smith, J. C. Yang, R. M. Sherry, R. E. Royal, U. S. Kammula, M. S. Hughes, T. E. Allen, C. L. Levy, et al. Prognostic Factors Related to Clinical Response in Patients with Metastatic Melanoma Treated by CTL-Associated Antigen-4 Blockade Clin. Cancer Res., November 15, 2007; 13(22): 6681 - 6688. [Abstract] [Full Text] [PDF]

				P. E. A. Souza, M. O. C. Rocha, C. A. S. Menezes, J. S. Coelho, A. C. L. Chaves, K. J. Gollob, and W. O. Dutra Trypanosoma cruzi Infection Induces Differential Modulation of Costimulatory Molecules and Cytokines by Monocytes and T Cells from Patients with Indeterminate and Cardiac Chagas' Disease Infect. Immun., April 1, 2007; 75(4): 1886 - 1894. [Abstract] [Full Text] [PDF]

				J. J. Engelhardt, T. J. Sullivan, and J. P. Allison CTLA-4 Overexpression Inhibits T Cell Responses through a CD28-B7-Dependent Mechanism J. Immunol., July 15, 2006; 177(2): 1052 - 1061. [Abstract] [Full Text] [PDF]

				D. Bernard, B. Riteau, J. D. Hansen, R. B. Phillips, F. Michel, P. Boudinot, and A. Benmansour Costimulatory Receptors in a Teleost Fish: Typical CD28, Elusive CTLA4 J. Immunol., April 1, 2006; 176(7): 4191 - 4200. [Abstract] [Full Text] [PDF]

				D. Homann, W. Dummer, T. Wolfe, E. Rodrigo, A. N. Theofilopoulos, M. B. A. Oldstone, and M. G. von Herrath Lack of Intrinsic CTLA-4 Expression Has Minimal Effect on Regulation of Antiviral T-Cell Immunity J. Virol., January 1, 2006; 80(1): 270 - 280. [Abstract] [Full Text] [PDF]

				P. Attia, G. Q. Phan, A. V. Maker, M. R. Robinson, M. M. Quezado, J. C. Yang, R. M. Sherry, S. L. Topalian, U. S. Kammula, R. E. Royal, et al. Autoimmunity Correlates With Tumor Regression in Patients With Metastatic Melanoma Treated With Anti-Cytotoxic T-Lymphocyte Antigen-4 J. Clin. Oncol., September 1, 2005; 23(25): 6043 - 6053. [Abstract] [Full Text] [PDF]

				P. J. Darlington, M. G. Kirchhof, G. Criado, J. Sondhi, and J. Madrenas Hierarchical Regulation of CTLA-4 Dimer-Based Lattice Formation and Its Biological Relevance for T Cell Inactivation J. Immunol., July 15, 2005; 175(2): 996 - 1004. [Abstract] [Full Text] [PDF]

				S. M. Anjos, M.-C. Tessier, and C. Polychronakos Association of the Cytotoxic T Lymphocyte-Associated Antigen 4 Gene with Type 1 Diabetes: Evidence for Independent Effects of Two Polymorphisms on the Same Haplotype Block J. Clin. Endocrinol. Metab., December 1, 2004; 89(12): 6257 - 6265. [Abstract] [Full Text] [PDF]

				H. Kwon, H.-S. Jun, L.-Y. Khil, and J.-W. Yoon Role of CTLA-4 in the Activation of Single- and Double-Positive Thymocytes J. Immunol., December 1, 2004; 173(11): 6645 - 6653. [Abstract] [Full Text] [PDF]

				N. Utku, A. Boerner, A. Tomschegg, F. Bennai-Sanfourche, G.-C. Bulwin, T. Heinemann, J. Loehler, R. S. Blumberg, and H.-D. Volk TIRC7 Deficiency Causes In Vitro and In Vivo Augmentation of T and B Cell Activation and Cytokine Response J. Immunol., August 15, 2004; 173(4): 2342 - 2352. [Abstract] [Full Text] [PDF]

				W. Stohl, D. Xu, K. S. Kim, C. S. David, and J. P. Allison MHC class II-independent and -dependent T cell expansion and B cell hyperactivity in vivo in mice deficient in CD152 (CTLA-4) Int. Immunol., July 1, 2004; 16(7): 895 - 904. [Abstract] [Full Text] [PDF]

				M Hirashima, T Fukazawa, K Abe, Y Morita, M Kusaoi, and H Hashimoto Expression and activity analyses of CTLA4 in peripheral blood lymphocytes in systemic lupus erythematosus patients Lupus, January 1, 2004; 13(1): 24 - 31. [Abstract] [PDF]

				G. Q. Phan, J. C. Yang, R. M. Sherry, P. Hwu, S. L. Topalian, D. J. Schwartzentruber, N. P. Restifo, L. R. Haworth, C. A. Seipp, L. J. Freezer, et al. Cancer regression and autoimmunity induced by cytotoxic T lymphocyte-associated antigen 4 blockade in patients with metastatic melanoma PNAS, July 8, 2003; 100(14): 8372 - 8377. [Abstract] [Full Text] [PDF]

				M. P. Pistillo, P. L. Tazzari, G. L. Palmisano, I. Pierri, A. Bolognesi, F. Ferlito, P. Capanni, L. Polito, M. Ratta, S. Pileri, et al. CTLA-4 is not restricted to the lymphoid cell lineage and can function as a target molecule for apoptosis induction of leukemic cells Blood, January 1, 2003; 101(1): 202 - 209. [Abstract] [Full Text] [PDF]

				A. M. Hall, F. J. Ward, M. A. Vickers, L.-M. Stott, S. J. Urbaniak, and R. N. Barker Interleukin-10-mediated regulatory T-cell responses to epitopes on a human red blood cell autoantigen Blood, December 15, 2002; 100(13): 4529 - 4536. [Abstract] [Full Text] [PDF]

				C. Guntermann and D. R. Alexander CTLA-4 Suppresses Proximal TCR Signaling in Resting Human CD4+ T Cells by Inhibiting ZAP-70 Tyr319 Phosphorylation: A Potential Role for Tyrosine Phosphatases J. Immunol., May 1, 2002; 168(9): 4420 - 4429. [Abstract] [Full Text] [PDF]

				J. S. Burr, S. L. Kimzey, D. R. Randolph, and J. M. Green CD28 and CTLA4 Coordinately Regulate Airway Inflammatory Cell Recruitment and T-Helper Cell Differentiation after Inhaled Allergen Am. J. Respir. Cell Mol. Biol., May 1, 2001; 24(5): 563 - 568. [Abstract] [Full Text]

				K. Runyon, K. Lee, K. Zuberek, M. Collins, J. P. Leonard, and K. Dunussi-Joannopoulos The combination of chemotherapy and systemic immunotherapy with soluble B7-immunoglobulin G leads to cure of murine leukemia and lymphoma and demonstration of tumor-specific memory responses Blood, April 15, 2001; 97(8): 2420 - 2426. [Abstract] [Full Text] [PDF]

				T. Iida, H. Ohno, C. Nakaseko, M. Sakuma, M. Takeda-Ezaki, H. Arase, E. Kominami, T. Fujisawa, and T. Saito Regulation of Cell Surface Expression of CTLA-4 by Secretion of CTLA-4-Containing Lysosomes Upon Activation of CD4+ T Cells J. Immunol., November 1, 2000; 165(9): 5062 - 5068. [Abstract] [Full Text] [PDF]

				M. L. Baroja, D. Luxenberg, T. Chau, V. Ling, C. A. Strathdee, B. M. Carreno, and J. Madrenas The Inhibitory Function of CTLA-4 Does Not Require Its Tyrosine Phosphorylation J. Immunol., January 1, 2000; 164(1): 49 - 55. [Abstract] [Full Text] [PDF]

				A. N. Schweitzer and A. H. Sharpe Mutual Regulation Between B7-1 (CD80) Expressed on T Cells and IL-4 J. Immunol., November 1, 1999; 163(9): 4819 - 4825. [Abstract] [Full Text] [PDF]

				S. Oki, T. Kohsaka, and M. Azuma Augmentation of CTLA-4 expression by wortmannin: involvement of lysosomal sorting properties of CTLA-4 Int. Immunol., September 1, 1999; 11(9): 1563 - 1571. [Abstract] [Full Text] [PDF]

				H. Schneider, M. Martin, F. A. Agarraberes, L. Yin, I. Rapoport, T. Kirchhausen, and C. E. Rudd Cytolytic T Lymphocyte-Associated Antigen-4 and the TCR{zeta}/CD3 Complex, But Not CD28, Interact with Clathrin Adaptor Complexes AP-1 and AP-2 J. Immunol., August 15, 1999; 163(4): 1868 - 1879. [Abstract] [Full Text] [PDF]

				F. J. Barrat, F. Le Deist, M. Benkerrou, P. Bousso, J. Feldmann, A. Fischer, and G. de Saint Basile Defective CTLA-4 cycling pathway in Chediak-Higashi syndrome: A possible mechanism for deregulation of T lymphocyte activation PNAS, July 20, 1999; 96(15): 8645 - 8650. [Abstract] [Full Text] [PDF]

				S. E Burastero and G. A Rossi Immunomodulation by interference with co-stimulatory molecules: therapeutic perspectives in asthma Thorax, June 1, 1999; 54(6): 554 - 557. [Full Text]

				M. C. Brunner, C. A. Chambers, F. K.-M. Chan, J. Hanke, A. Winoto, and J. P. Allison CTLA-4-Mediated Inhibition of Early Events of T Cell Proliferation J. Immunol., May 15, 1999; 162(10): 5813 - 5820. [Abstract] [Full Text] [PDF]

				C.A. CHAMBERS and J.P. ALLISON CTLA-4 -- The Costimulatory Molecule That Doesn't: Regulation of T-cell Responses by Inhibition Cold Spring Harb Symp Quant Biol, January 1, 1999; 64(0): 303 - 312. [Abstract] [PDF]

				G. Boulougouris, J. D. McLeod, Y. I. Patel, C. N. Ellwood, L. S. K. Walker, and D. M. Sansom Positive and Negative Regulation of Human T Cell Activation Mediated by the CTLA-4/CD28 Ligand CD80 J. Immunol., October 15, 1998; 161(8): 3919 - 3924. [Abstract] [Full Text] [PDF]

				M. L. Murphy, S. E. J. Cotterell, P. M. A. Gorak, C. R. Engwerda, and P. M. Kaye Blockade of CTLA-4 Enhances Host Resistance to the Intracellular Pathogen, Leishmania donovani J. Immunol., October 15, 1998; 161(8): 4153 - 4160. [Abstract] [Full Text] [PDF]

				M.-L. Alegre, H. Shiels, C. B. Thompson, and T. F. Gajewski Expression and Function of CTLA-4 in Th1 and Th2 Cells J. Immunol., October 1, 1998; 161(7): 3347 - 3356. [Abstract] [Full Text] [PDF]

				A. N. Schweitzer and A. H. Sharpe Studies Using Antigen-Presenting Cells Lacking Expression of Both B7-1 (CD80) and B7-2 (CD86) Show Distinct Requirements for B7 Molecules During Priming Versus Restimulation of Th2 But Not Th1 Cytokine Production J. Immunol., September 15, 1998; 161(6): 2762 - 2771. [Abstract] [Full Text] [PDF]

				D. Cefai, H. Schneider, O. Matangkasombut, H. Kang, J. Brody, and C. E. Rudd CD28 Receptor Endocytosis Is Targeted by Mutations That Disrupt Phosphatidylinositol 3-Kinase Binding and Costimulation J. Immunol., March 1, 1998; 160(5): 2223 - 2230. [Abstract] [Full Text] [PDF]

				P. J. Blair, J. L. Riley, B. L. Levine, K. P. Lee, N. Craighead, T. Francomano, S. J. Perfetto, G. S. Gray, B. M. Carreno, and C. H. June Cutting Edge: CTLA-4 Ligation Delivers a Unique Signal to Resting Human CD4 T Cells That Inhibits Interleukin-2 Secretion but Allows Bcl-XL Induction J. Immunol., January 1, 1998; 160(1): 12 - 15. [Abstract] [Full Text] [PDF]

				F. Colucci, M.-L. Bergman, C. Penha-Goncalves, C. M. Cilio, and D. Holmberg Apoptosis resistance of nonobese diabetic peripheral lymphocytes linked to the Idd5 diabetes susceptibility region PNAS, August 5, 1997; 94(16): 8670 - 8674. [Abstract] [Full Text] [PDF]

				H. T. Leung, J. Bradshaw, J. S. Cleaveland, and P. S. Linsley 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., October 20, 1995; 270(42): 25107 - 25114. [Abstract] [Full Text] [PDF]

				P. S. Linsley, S. G. Nadler, Jür. Bajorath, R. Peach, H. T. Leung, J. Rogers, J. Bradshaw, M. Stebbins, G. Leytze, W. Brady, et al. Binding Stoichiometry of the Cytotoxic T Lymphocyte-associated Molecule-4 (CTLA-4) J. Biol. Chem., June 23, 1995; 270(25): 15417 - 15424. [Abstract] [Full Text] [PDF]