title: Chaperone activity of α B-crystallin is responsible for its incorrect assignment as an autoantigen in multiple sclerosis.
abstract: For 15 y, α B-crystallin (heat shock protein [Hsp] B5) has been labeled an autoantigen in multiple sclerosis (MS) based on humoral and cellular responses found in humans and animal models. However, there have been several scientific inconsistencies with this assignment, ranging from studies demonstrating small differences in anticrystallin responses between patients and healthy individuals to the inability of crystallin-specific T cells to induce symptoms of experimental allergic encephalomyelitis in animal models. Experiments in this article demonstrate that the putative anti-HspB5 Abs from 23 MS patients cross-react with 7 other members of the human small Hsp family and were equally present in normal plasma. Biolayer interferometry demonstrates that the binding was temperature dependent, and that the calculated K(a) increased as the concentration of the sHsp decreased. These two patterns are characteristic of multiple binding sites with varying affinities, the composition of which changes with temperature, supporting the hypothesis that HspB5 bound the Ab and not the reverse. HspB5 also precipitated Ig heavy and L chains from sera from patients with MS. These results establish that small Hsps bind Igs with high affinity and refute much of the serological data used to assign α B-crystallin as an autoantigen.
authors.affiliation: Division of Immunology and Rheumatology, Department of Medicine, Stanford University School of Medicine, Stanford University, Stanford, CA 94305, USA.
authors.forename: Jonathan B
authors.initials: JB
authors.lastname: Rothbard
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Zhao
authors.forename: Orr
authors.initials: O
authors.lastname: Sharpe
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Strohman
authors.forename: Michael
authors.initials: M
authors.lastname: Kurnellas
authors.forename: Elizabeth D
authors.initials: ED
authors.lastname: Mellins
authors.forename: William H
authors.initials: WH
authors.lastname: Robinson
authors.forename: Lawrence
authors.initials: L
authors.lastname: Steinman
mesh_terms: D000818:Animals; D001324:Autoantigens / Q000276:immunology / Q000378:metabolism; D001666:Binding Sites, Antibody; D004195:Disease Models, Animal; D050888:Heat-Shock Proteins, Small / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D018832:Molecular Chaperones / Q000737:chemistry / Q000378:metabolism*; D009103:Multiple Sclerosis / Q000276:immunology / Q000378:metabolism; D038203:alpha-Crystallin B Chain / Q000737:chemistry / Q000378:metabolism*
chemical_list: D001324:Autoantigens; D050888:Heat-Shock Proteins, Small; D007141:Immunoglobulin Fc Fragments; D018832:Molecular Chaperones; D038203:alpha-Crystallin B Chain