title: Rabbit anti-HMG-17 antibodies recognize similar epitopes on the HMG-17 molecule as lupus autoantibodies. Relation with histone H1 defined epitopes.
abstract: HMG-17 is a nucleosomal protein which is an immune target of autoantibodies in systemic lupus erythematosus (SLE) and other autoimmune diseases. Autoantibody production in SLE is believed to result from autoantigen specific immune stimulation and subsequently, it is expected that antigenic determinants recognized by SLE autoantibodies and induced antibodies by immunization are quite similar. To examine this issue, rabbits were immunized with purified HMG-17. The produced antiserum showed cross reactivity on blots and in inhibition ELISA with histone H1, even after its affinity purification with immobilized HMG-17. Finally, purification of the antiserum over H1 absorbed on nitrocellulose membrane produced specific anti-HMG-17 antibodies in the supernatant and anti-HMG-17/H1 antibodies that were bound to H1. SLE sera positive for HMG-17 had also cross reactivity with H1, and following the same procedure as before we received HMG-17 specific SLE autoantibodies and anti-HMG-17/H1 autoantibodies. Using the multipin epitope mapping technology, 19 overlapping 15-mer HMG-17 peptides and six 15-peptides, corresponding to known epitopes of histone H1, were synthesized. Four major epitopes were identified on the HMG-17 molecule, reactive with induced anti-HMG-17 antibodies, and these were the same as major autoepitopes In SLE. The sequence 25-51 of HMG-17, part of its DNA-binding domain, was recognized by the anti-HMG-17/H1 antibodies that were bound to H1. These antibodies recognized also defined epitopes of H1. Our results show that SLE autoantibodies can be directed against the same or similar epitopes as do IgGs evoked during the active immunization of animals, and provide additional evidence that autosensitization with an autoantigen might be operative. The possibility that the same or similar epitopes are found on different molecules (in this study HMG-17 and H1) supports the fact that there are rules by which nature selects the most dominant immunodeterminant to a given protein, which often represents functional or structural sites in the autoantigen.
authors.affiliation: Laboratory of Biological Chemistry, University of Ioannina, Medical School, 451 10 Ioannina, Greece.
authors.forename: Vassiliki A
authors.initials: VA
authors.lastname: Boumba
authors.forename: Konstantin
authors.initials: K
authors.lastname: Seferiadis
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D001323:Autoantibodies / Q000276:immunology*; D002417:Cattle; D018604:Epitope Mapping; D000939:Epitopes / Q000276:immunology*; D024242:HMGN2 Protein / Q000138:chemical synthesis / Q000235:genetics / Q000276:immunology*; D006657:Histones / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D008180:Lupus Erythematosus, Systemic / Q000097:blood / Q000276:immunology*; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000235:genetics / Q000276:immunology; D011817:Rabbits; D013552:Swine; D013950:Thymus Gland / Q000737:chemistry
chemical_list: D001323:Autoantibodies; D000939:Epitopes; D024242:HMGN2 Protein; D006657:Histones; D010455:Peptides