Cleavage by Granzyme B Is Strongly Predictive of Autoantigen Status: Implications for Initiation of Autoimmunity

Systemic autoimmune diseases are a genetically complex, heterogeneous group of disorders in which the immune system targets a diverse but highly specific group of intracellular autoantigens. The molecules targeted are not unified by common structure, function, or distribution in control cells but become clustered and concentrated in surface blebs when cells undergo apoptosis. We show here that the majority of autoantigens targeted across the spectrum of human systemic autoimmune diseases are efficiently cleaved by granzyme B in vitro and during cytotoxic lymphocyte granule–induced death, generating unique fragments not observed during any other form of apoptosis. These molecules are not cleaved by caspase-8, although this protease has a very similar specificity to granzyme B. The granzyme B cleavage sites in autoantigens contain amino acids in the P₂ and P₃ positions that are preferred by granzyme B but are not tolerated by caspase-8. In contrast to autoantigens, nonautoantigens are either not cleaved by granzyme B or are cleaved to generate fragments identical to those formed in other forms of apoptosis. The striking ability of granzyme B to generate unique fragments is therefore an exclusive property of autoantigens and unifies the majority of molecules targeted in this spectrum of diseases. These results focus attention on the role of the cytotoxic lymphocyte granule–induced death pathway in the initiation and propagation of systemic autoimmunity.

Key Words: apoptosis, cytotoxic T lymphocyte, protease, caspase

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

• Park, S. G., Schimmel, P., Kim, S. (2008). Aminoacyl tRNA synthetases and their connections to disease. Proc. Natl. Acad. Sci. USA 105: 11043-11049 [Abstract] [Full Text]  
• Eggleton, P., Haigh, R., Winyard, P. G. (2008). Consequence of neo-antigenicity of the 'altered self'. Rheumatology (Oxford) 47: 567-571 [Abstract] [Full Text]  
• Casciola-Rosen, L., Garcia-Calvo, M., Bull, H. G., Becker, J. W., Hines, T., Thornberry, N. A., Rosen, A. (2007). Mouse and Human Granzyme B Have Distinct Tetrapeptide Specificities and Abilities to Recruit the Bid Pathway. J. Biol. Chem. 282: 4545-4552 [Abstract] [Full Text]  
• Wehner, J., Morrell, C. N., Reynolds, T., Rodriguez, E. R., Baldwin, W. M. III (2007). Antibody and Complement in Transplant Vasculopathy. Circ. Res. 100: 191-203 [Abstract] [Full Text]  
• Ganor, Y., Teichberg, V. I., Levite, M. (2007). TCR Activation Eliminates Glutamate Receptor GluR3 from the Cell Surface of Normal Human T Cells, via an Autocrine/Paracrine Granzyme B-Mediated Proteolytic Cleavage. J. Immunol. 178: 683-692 [Abstract] [Full Text]  
• Grader-Beck, T., Casciola-Rosen, L., Lang, T. J., Puliaev, R., Rosen, A., Via, C. S. (2007). Apoptotic Splenocytes Drive the Autoimmune Response to Poly(ADP-ribose) Polymerase 1 in a Murine Model of Lupus. J. Immunol. 178: 95-102 [Abstract] [Full Text]  
• Specks, U., Fass, D. N., Finkielman, J. D., Hummel, A. M., Viss, M. A., Litwiller, R. D., Mcdonald, C. J. (2007). Functional Significance of Asn-linked Glycosylation of Proteinase 3 for Enzymatic Activity, Processing, Targeting, and Recognition by Anti-neutrophil Cytoplasmic Antibodies. J Biochem 141: 101-112 [Abstract] [Full Text]  
• Bell, C. W., Jiang, W., Reich, C. F. III, Pisetsky, D. S. (2006). The extracellular release of HMGB1 during apoptotic cell death. Am. J. Physiol. Cell Physiol. 291: C1318-C1325 [Abstract] [Full Text]  
• Loeb, C. R. K., Harris, J. L., Craik, C. S. (2006). Granzyme B Proteolyzes Receptors Important to Proliferation and Survival, Tipping the Balance toward Apoptosis. J. Biol. Chem. 281: 28326-28335 [Abstract] [Full Text]  
• Xiong, Z, Shaibani, A, Li, Y-P, Yan, Y, Zhang, S, Yang, Y, Yang, F, Wang, H, Yang, X-F (2006). Alternative splicing factor ASF/SF2 is down regulated in inflamed muscle. J. Clin. Pathol. 59: 855-861 [Abstract] [Full Text]  
• Denny, M. F., Chandaroy, P., Killen, P. D., Caricchio, R., Lewis, E. E., Richardson, B. C., Lee, K.-D., Gavalchin, J., Kaplan, M. J. (2006). Accelerated Macrophage Apoptosis Induces Autoantibody Formation and Organ Damage in Systemic Lupus Erythematosus. J. Immunol. 176: 2095-2104 [Abstract] [Full Text]  
• Bredberg, A., Henriksson, G., Larsson, A., Manthorpe, R., Sallmyr, A. (2005). Sjogren's syndrome and the danger model. Rheumatology (Oxford) 44: 965-970 [Full Text]  
• Riemekasten, G., Hahn, B. H. (2005). Key autoantigens in SLE. Rheumatology (Oxford) 44: 975-982 [Full Text]  
• Comtesse, N., Zippel, A., Walle, S., Monz, D., Backes, C., Fischer, U., Mayer, J., Ludwig, N., Hildebrandt, A., Keller, A., Steudel, W.-I., Lenhof, H.-P., Meese, E. (2005). Complex humoral immune response against a benign tumor: Frequent antibody response against specific antigens as diagnostic targets. Proc. Natl. Acad. Sci. USA 102: 9601-9606 [Abstract] [Full Text]  
• Backes, C., Kuentzer, J., Lenhof, H.-P., Comtesse, N., Meese, E. (2005). GraBCas: a bioinformatics tool for score-based prediction of Caspase- and Granzyme B-cleavage sites in protein sequences. Nucleic Acids Res 33: W208-W213 [Abstract] [Full Text]  
• Moss, C. X., Matthews, S. P., Lamont, D. J., Watts, C. (2005). Asparagine Deamidation Perturbs Antigen Presentation on Class II Major Histocompatibility Complex Molecules. J. Biol. Chem. 280: 18498-18503 [Abstract] [Full Text]  
• Goldbach-Mansky, R, Suson, S, Wesley, R, Hack, C E, El-Gabalawy, H S, Tak, P P (2005). Raised granzyme B levels are associated with erosions in patients with early rheumatoid factor positive rheumatoid arthritis. Ann Rheum Dis 64: 715-721 [Abstract] [Full Text]  
• Bien, C. G., Granata, T., Antozzi, C., Cross, J. H., Dulac, O., Kurthen, M., Lassmann, H., Mantegazza, R., Villemure, J.-G., Spreafico, R., Elger, C. E. (2005). Pathogenesis, diagnosis and treatment of Rasmussen encephalitis: A European consensus statement. Brain 128: 454-471 [Abstract] [Full Text]  
• Sebbagh, M., Hamelin, J., Bertoglio, J., Solary, E., Breard, J. (2005). Direct cleavage of ROCK II by granzyme B induces target cell membrane blebbing in a caspase-independent manner. JEM 201: 465-471 [Abstract] [Full Text]  
• Mahrus, S., Kisiel, W., Craik, C. S. (2004). Granzyme M Is a Regulatory Protease That Inactivates Proteinase Inhibitor 9, an Endogenous Inhibitor of Granzyme B. J. Biol. Chem. 279: 54275-54282 [Abstract] [Full Text]  
• Gabler, C, Blank, N, Hieronymus, T, Schiller, M, Berden, J H M, Kalden, J R, Lorenz, H-M (2004). Extranuclear detection of histones and nucleosomes in activated human lymphoblasts as an early event in apoptosis. Ann Rheum Dis 63: 1135-1144 [Abstract] [Full Text]  
• Graham, K. L., Gustin, K. E., Rivera, C., Kuyumcu-Martinez, N. M., Choe, S. S., Lloyd, R. E., Sarnow, P., Utz, P. J. (2004). Proteolytic Cleavage of the Catalytic Subunit of DNA-Dependent Protein Kinase during Poliovirus Infection. J. Virol. 78: 6313-6321 [Abstract] [Full Text]  
• Rosen, A., Casciola-Rosen, L. (2004). ALTERED AUTOANTIGEN STRUCTURE IN SJoGREN'S SYNDROME: IMPLICATIONS FOR THE PATHOGENESIS OF AUTOIMMUNE TISSUE DAMAGE. CROBM 15: 156-164 [Abstract] [Full Text]  
• Sun, J., Bird, C. H., Thia, K. Y., Matthews, A. Y., Trapani, J. A., Bird, P. I. (2004). Granzyme B Encoded by the Commonly Occurring Human RAH Allele Retains Pro-apoptotic Activity. J. Biol. Chem. 279: 16907-16911 [Abstract] [Full Text]  
• Russell, A. I., Cunninghame Graham, D. S., Shepherd, C., Roberton, C. A., Whittaker, J., Meeks, J., Powell, R. J., Isenberg, D. A., Walport, M. J., Vyse, T. J. (2004). Polymorphism at the C-reactive protein locus influences gene expression and predisposes to systemic lupus erythematosus. Hum Mol Genet 13: 137-147 [Abstract] [Full Text]  
• Greidinger, E. L., Foecking, M. F., Magee, J., Wilson, L., Ranatunga, S., Ortmann, R. A., Hoffman, R. W. (2004). A Major B Cell Epitope Present on the Apoptotic but Not the Intact Form of the U1-70-kDa Ribonucleoprotein Autoantigen. J. Immunol. 172: 709-716 [Abstract] [Full Text]  
• Caricchio, R., McPhie, L., Cohen, P. L. (2003). Ultraviolet B Radiation-Induced Cell Death: Critical Role of Ultraviolet Dose in Inflammation and Lupus Autoantigen Redistribution. J. Immunol. 171: 5778-5786 [Abstract] [Full Text]  
• Ulanet, D. B., Torbenson, M., Dang, C. V., Casciola-Rosen, L., Rosen, A. (2003). Unique conformation of cancer autoantigen B23 in hepatoma: A mechanism for specificity in the autoimmune response. Proc. Natl. Acad. Sci. USA 100: 12361-12366 [Abstract] [Full Text]  
• Andrade, F., Casciola-Rosen, L. A., Rosen, A. (2003). A Novel Domain in Adenovirus L4-100K Is Required for Stable Binding and Efficient Inhibition of Human Granzyme B: Possible Interaction with a Species-Specific Exosite. Mol. Cell. Biol. 23: 6315-6326 [Abstract] [Full Text]  
• Mok, C C, Lau, C S (2003). Pathogenesis of systemic lupus erythematosus. J. Clin. Pathol. 56: 481-490 [Abstract] [Full Text]  
• D'Agnillo, P., Levine, J. S., Subang, R., Rauch, J. (2003). Prothrombin Binds to the Surface of Apoptotic, But Not Viable, Cells and Serves as a Target of Lupus Anticoagulant Autoantibodies. J. Immunol. 170: 3408-3422 [Abstract] [Full Text]  
• Charles, P J (2003). Defective waste disposal: does it induce autoantibodies in SLE?. Ann Rheum Dis 62: 1-3 [Full Text]  
• Ascherman, D. P., Oriss, T. B., Oddis, C. V., Wright, T. M. (2002). Critical Requirement for Professional APCs in Eliciting T Cell Responses to Novel Fragments of Histidyl-tRNA Synthetase (Jo-1) in Jo-1 Antibody-Positive Polymyositis. J. Immunol. 169: 7127-7134 [Abstract] [Full Text]  
• Dieker, J W., van der Vlag, J, Berden, J H. (2002). Triggers for anti-chromatin autoantibody production in SLE. Lupus 11: 856-864 [Abstract]  
• Kamachi, M., Le, T. M., Kim, S. J., Geiger, M. E., Anderson, P., Utz, P. J. (2002). Human Autoimmune Sera as Molecular Probes for the Identification of an Autoantigen Kinase Signaling Pathway. JEM 196: 1213-1226 [Abstract] [Full Text]  
• Thebault, S., Gilbert, D., Hubert, M., Drouot, L., Machour, N., Lange, C., Charlionet, R., Tron, F. (2002). Orderly Pattern of Development of the Autoantibody Response in (New Zealand White x BXSB)F1 Lupus Mice: Characterization of Target Antigens and Antigen Spreading by Two-Dimensional Gel Electrophoresis and Mass Spectrometry. J. Immunol. 169: 4046-4053 [Abstract] [Full Text]  
• Lee, K.-J., Dong, X., Wang, J., Takeda, Y., Dynan, W. S. (2002). Identification of Human Autoantibodies to the DNA Ligase IV/XRCC4 Complex and Mapping of an Autoimmune Epitope to a Potential Regulatory Region. J. Immunol. 169: 3413-3421 [Abstract] [Full Text]  
• Greidinger, E. L., Foecking, M. F., Schafermeyer, K. R., Bailey, C. W., Primm, S. L., Lee, D. R., Hoffman, R. W. (2002). T Cell Immunity in Connective Tissue Disease Patients Targets the RNA Binding Domain of the U1-70kDa Small Nuclear Ribonucleoprotein. J. Immunol. 169: 3429-3437 [Abstract] [Full Text]  
• Hansen, M. H., Nielsen, H. V., Ditzel, H. J. (2002). Translocation of an Intracellular Antigen to the Surface of Medullary Breast Cancer Cells Early in Apoptosis Allows for an Antigen-Driven Antibody Response Elicited by Tumor-Infiltrating B Cells. J. Immunol. 169: 2701-2711 [Abstract] [Full Text]  
• Clancy, R. M., Askanase, A. D., Kapur, R. P., Chiopelas, E., Azar, N., Miranda-Carus, M. E., Buyon, J. P. (2002). Transdifferentiation of Cardiac Fibroblasts, a Fetal Factor in Anti-SSA/Ro-SSB/La Antibody-Mediated Congenital Heart Block. J. Immunol. 169: 2156-2163 [Abstract] [Full Text]  
• Chernysheva, A. D., Kirou, K. A., Crow, M. K. (2002). T Cell Proliferation Induced by Autologous Non-T Cells Is a Response to Apoptotic Cells Processed by Dendritic Cells. J. Immunol. 169: 1241-1250 [Abstract] [Full Text]  
• Stuart, L., Hughes, J. (2002). Apoptosis and autoimmunity. Nephrol Dial Transplant 17: 697-700 [Full Text]  
• Du, L., Fukushima, S., Sallmyr, A., Manthorpe, R., Bredberg, A. (2002). Exposure of HEp-2 Cells to Stress Conditions Influences Antinuclear Antibody Reactivity. CVI 9: 287-294 [Abstract] [Full Text]  
• McHugh, N. J. (2002). Systemic lupus erythematosus and dysregulated apoptosis--what is the evidence?. Rheumatology (Oxford) 41: 242-245 [Full Text]  
• Castiglioni, P., Martin-Fontecha, A., Milan, G., Tomajer, V., Magni, F., Michaelsson, J., Rugarli, C., Rosato, A., Bellone, M. (2002). Apoptosis-dependent Subversion of the T-Lymphocyte Epitope Hierarchy in Lymphoma Cells. Cancer Res. 62: 1116-1122 [Abstract] [Full Text]  
• Zhang, Y., O'Brien, B., Trudeau, J., Tan, R., Santamaria, P., Dutz, J. P. (2002). In Situ {beta} Cell Death Promotes Priming of Diabetogenic CD8 T Lymphocytes. J. Immunol. 168: 1466-1472 [Abstract] [Full Text]  
• Shoshan, Y., Shapira, I., Toubi, E., Frolkis, I., Yaron, M., Mevorach, D. (2001). Accelerated Fas-Mediated Apoptosis of Monocytes and Maturing Macrophages from Patients with Systemic Lupus Erythematosus: Relevance to In Vitro Impairment of Interaction with iC3b-Opsonized Apoptotic Cells. J. Immunol. 167: 5963-5969 [Abstract] [Full Text]  
• Kuwana, M., Okano, T., Ogawa, Y., Kaburaki, J., Kawakami, Y. (2001). Autoantibodies to the Amino-Terminal Fragment of {beta}-Fodrin Expressed in Glandular Epithelial Cells in Patients with Sjogren's Syndrome. J. Immunol. 167: 5449-5456 [Abstract] [Full Text]  
• Hansen, M. H., Nielsen, H., Ditzel, H. J. (2001). The tumor-infiltrating B cell response in medullary breast cancer is oligoclonal and directed against the autoantigen actin exposed on the surface of apoptotic cancer cells. Proc. Natl. Acad. Sci. USA 10.1073/pnas.171460798v1 [Abstract] [Full Text]  
• Smyth, M. J., Kelly, J. M., Sutton, V. R., Davis, J. E., Browne, K. A., Sayers, T. J., Trapani, J. A. (2001). Unlocking the secrets of cytotoxic granule proteins. J. Leukoc. Biol. 70: 18-29 [Abstract] [Full Text]  
• Haque, M. A., Hawes, J. W., Blum, J. S. (2001). Cysteinylation of MHC Class II Ligands: Peptide Endocytosis and Reduction Within APC Influences T Cell Recognition. J. Immunol. 166: 4543-4551 [Abstract] [Full Text]  
• Ratovitski, E. A., Patturajan, M., Hibi, K., Trink, B., Yamaguchi, K., Sidransky, D. (2001). p53 associates with and targets Delta Np63 into a protein degradation pathway. Proc. Natl. Acad. Sci. USA 98: 1817-1822 [Abstract] [Full Text]  
• Gahring, L. C., Carlson, N. G., Meyer, E. L., Rogers, S. W. (2001). Cutting Edge: Granzyme B Proteolysis of a Neuronal Glutamate Receptor Generates an Autoantigen and Is Modulated by Glycosylation. J. Immunol. 166: 1433-1438 [Abstract] [Full Text]  
• Maraia, R. J., Intine, R. V. A. (2001). Recognition of Nascent RNA by the Human La Antigen: Conserved and Divergent Features of Structure and Function. Mol. Cell. Biol. 21: 367-379 [Full Text]  
• Zampieri, S, Degen, W, Ghiradello, A, Doria, A, van Venrooij, W J (2001). Dephosphorylation of autoantigenic ribosomal P proteins during Fas-L induced apoptosis: a possible trigger for the development of the autoimmune response in patients with systemic lupus erythematosus. Ann Rheum Dis 60: 72-76 [Abstract] [Full Text]  
• Rovere, P., Peri, G., Fazzini, F., Bottazzi, B., Doni, A., Bondanza, A., Zimmermann, V. S., Garlanda, C., Fascio, U., Sabbadini, M. G., Rugarli, C., Mantovani, A., Manfredi, A. A. (2000). The long pentraxin PTX3 binds to apoptotic cells and regulates their clearance by antigen-presenting dendritic cells. Blood 96: 4300-4306 [Abstract] [Full Text]  
• Taylor, P. R., Carugati, A., Fadok, V. A., Cook, H. T., Andrews, M., Carroll, M. C., Savill, J. S., Henson, P. M., Botto, M., Walport, M. J. (2000). A Hierarchical Role for Classical Pathway Complement Proteins in the Clearance of Apoptotic Cells In Vivo. JEM 192: 359-366 [Abstract] [Full Text]  
• Mancini, M., Machamer, C. E., Roy, S., Nicholson, D. W., Thornberry, N. A., Casciola-Rosen, L. A., Rosen, A. (2000). Caspase-2 Is Localized at the Golgi Complex and Cleaves Golgin-160 during Apoptosis. JCB 149: 603-612 [Abstract] [Full Text]  
• Freed, J. H., Marrs, A., VanderWall, J., Cohen, P. L., Eisenberg, R. A. (2000). MHC Class II-Bound Self Peptides from Autoimmune MRL/lpr Mice Reveal Potential T Cell Epitopes for Autoantibody Production in Murine Systemic Lupus Erythematosus. J. Immunol. 164: 4697-4705 [Abstract] [Full Text]  
• Sun, J., Whisstock, J. C., Harriott, P., Walker, B., Novak, A., Thompson, P. E., Smith, A. I., Bird, P. I. (2001). Importance of the P4' Residue in Human Granzyme B Inhibitors and Substrates Revealed by Scanning Mutagenesis of the Proteinase Inhibitor 9 Reactive Center Loop. J. Biol. Chem. 276: 15177-15184 [Abstract] [Full Text]  
• Hansen, M. H., Nielsen, H., Ditzel, H. J. (2001). The tumor-infiltrating B cell response in medullary breast cancer is oligoclonal and directed against the autoantigen actin exposed on the surface of apoptotic cancer cells. Proc. Natl. Acad. Sci. USA 98: 12659-12664 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search

TABLE OF CONTENTS