title: Specific immunogenicity of heat shock protein gp96 derives from chaperoned antigenic peptides and not from contaminating proteins.
abstract: The peptide-binding property of MHC is central to adaptive immunological functions. A similar property of heat shock proteins (HSPs) hsp70 and hsp90 has been implicated in Ag presentation by MHC and in cross-priming. The peptide-binding pocket of hsp70 has been characterized structurally and functionally and a peptide-binding site in gp96 (of hsp90 family) has been defined. Nonetheless, questions persist whether the specific immunogenicity of HSP preparations derives from the peptides chaperoned by the HSPs or by proteins contaminating the HSP preparations. Because absolute purity of a protein preparation is a metaphysical concept, other approaches are necessary to address the question. In this study, we demonstrate that the specific immunogenicity of gp96 preparations isolated from cells expressing beta-galactosidase derives from the MHC I epitope precursors associated with the gp96 and not from contaminating beta-galactosidase protein nor unassociated fragments derived from it. Although the observations here are limited to a single HSP and antigenic peptides chaperoned by it, they can be extended broadly.
authors.affiliation: Department of Immunology, University of Pittsburgh, Pittsburgh, PA 15261, USA. rjb42@pitt.edu
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Binder
authors.forename: John B
authors.initials: JB
authors.lastname: Kelly
authors.forename: Ralph E
authors.initials: RE
authors.lastname: Vatner
authors.forename: Pramod K
authors.initials: PK
authors.lastname: Srivastava
mesh_terms: D000818:Animals; D017951:Antigen Presentation / Q000276:immunology; D000951:Antigens, Neoplasm / Q000276:immunology* / Q000302:isolation & purification; D002460:Cell Line; D000939:Epitopes / Q000276:immunology; D005260:Female; D018841:HSP90 Heat-Shock Proteins / Q000276:immunology*; D015395:Histocompatibility Antigens Class I / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D010455:Peptides / Q000276:immunology*; D001616:beta-Galactosidase / Q000235:genetics / Q000276:immunology*
chemical_list: D000951:Antigens, Neoplasm; D000939:Epitopes; D018841:HSP90 Heat-Shock Proteins; D015395:Histocompatibility Antigens Class I; D010455:Peptides; C052646:sarcoma glycoprotein gp96 rejection antigens; D001616:beta-Galactosidase