title: Presentation of the Goodpasture autoantigen requires proteolytic unlocking steps that destroy prominent T cell epitopes.
abstract: The most abundant autoreactive T cells in patients with Goodpasture's disease are specific for peptides in the autoantigen that have high affinity for the disease-associated HLA class II molecule, DR15. How can such T cells escape self-tolerance mechanisms? This study showed that these peptides are highly susceptible to destruction in the earliest stages of antigen processing, and some must be cleaved for antigen digestion to be possible ("unlocking"). Goodpasture autoantigen [collagen alpha3(IV)NC1; approximately 31 kD] that was incubated with B cell lysosomes was cleaved within a few minutes to form approximately 9- and approximately 22-kD fragments, then increasing quantities of smaller peptides. The processing was completely abrogated by pepstatin A, a specific inhibitor of cathepsin D/E, even though lysosomal extracts contain a rich array of proteases. Purified cathepsin D generated the same major alpha3(IV)NC1 fragments as entire lysosomes, suggesting that cathepsin D cleavages are required to initiate alpha3(IV)NC1 processing. The initial unlocking cleavages destroyed two major self-epitopes, and subsequent preferred cleavages destroyed all of the other T cell epitopes that are recognized by most patients' autoreactive T cells. The responses of T cell clones that are specific for a major disease-associated peptide to antigen-pulsed intact antigen-presenting cells were substantially enhanced by pepstatin A treatment. Therefore, cathepsin D activity significantly diminishes presentation of alpha3(IV)NC1 peptides that are recognized by patients' T cells by destroying the peptides in early processing. These observations can explain why the mature T cell repertoire includes reactivity toward these self-peptides and suggests that a key factor in disease initiation is likely to be a shift in antigen processing.
authors.affiliation: MRC Centre for Inflammation Research (Renal Autoimmunity), University of Edinburgh, Edinburgh, UK.
authors.forename: Juan
authors.initials: J
authors.lastname: Zou
authors.forename: Lorna
authors.initials: L
authors.lastname: Henderson
authors.forename: Vicky
authors.initials: V
authors.lastname: Thomas
authors.forename: Patricia
authors.initials: P
authors.lastname: Swan
authors.forename: A Neil
authors.initials: AN
authors.lastname: Turner
authors.forename: Richard G
authors.initials: RG
authors.lastname: Phelps
mesh_terms: D019867:Anti-Glomerular Basement Membrane Disease; D000938:Antigen-Presenting Cells / Q000276:immunology*; D016282:Aspartic Acid Endopeptidases / Q000276:immunology / Q000378:metabolism*; D001324:Autoantigens / Q000187:drug effects / Q000378:metabolism*; D002402:Cathepsin D / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D020059:Cathepsin E / Q000037:antagonists & inhibitors; D024141:Collagen Type IV / Q000187:drug effects / Q000378:metabolism*; D000939:Epitopes / Q000276:immunology; D006801:Humans; D008247:Lysosomes / Q000201:enzymology / Q000502:physiology; D010436:Pepstatins / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000187:drug effects / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D001324:Autoantigens; D024141:Collagen Type IV; D000939:Epitopes; D010436:Pepstatins; C057112:type IV collagen alpha3 chain; C119090:CERT1 protein, human; D017346:Protein Serine-Threonine Kinases; D016282:Aspartic Acid Endopeptidases; D020059:Cathepsin E; D002402:Cathepsin D; C031375:pepstatin