title: Autoantibodies recognizing the amino terminal 1-17 segment of CENP-A display unique specificities in systemic sclerosis.
abstract: Centromere-associated protein A (CENP-A), a common autoimmune target in a subset of systemic sclerosis patients, appears to have no role to explain why its corresponding auto-antibodies are more frequently found in the limited than the diffuse form of systemic sclerosis. Therefore, we investigated the fine specificity of anti-CENP-A antibodies as a first step to understanding their role in systemic sclerosis pathology. We focused on the amino-terminal portion of CENP-A spanning amino acids 1 to 17 (Ap(1-17)), which represents, along with Ap(17-30), an immunodominant epitope of the protein. Peptide Ap(1-17) was used to purify antibodies from 8 patients with systemic sclerosis. Anti-Ap(1-17) antibodies specifically reacted with human CENP-A but did not cross-react with CENP-B or Ap(17-30). Panning of a phage display peptide library with anti-Ap(1-17) antibodies from 2 patients identified two novel, partially overlapping motifs, <(5)Rx(st)xKP(10)> and <(9)KPxxPxR(15)> as the result of the alignment of specific phage clone insert sequences. Anti-Ap(1-17) IgG from the 8 patients had different reactivities to isolated phage clone insert sequences. Scanning the Swiss-Prot database revealed a large number of different types of proteins containing the two Ap(1-17) antigenic motifs. These data show that anti-CENP-A(1-17) antibodies are generated independently from anti-CENP-B antibodies and display great heterogeneity in their specificity by recognizing different motifs within that peptide sequence. This finding, along with the widespread interspecies and human tissue distribution of the two motifs, suggests that the number of motif-expressing proteins which can be the potential target of these antibodies is markedly higher than that estimated from the peptide-based epitope spreading model.
authors.affiliation: Department of Internal Medicine (DIMO), University of Bari Medical School, Bari, Italy.
authors.forename: Elvira
authors.initials: E
authors.lastname: Favoino
authors.forename: Liboria
authors.initials: L
authors.lastname: Digiglio
authors.forename: Giovanna
authors.initials: G
authors.lastname: Cuomo
authors.forename: Isabella E
authors.initials: IE
authors.lastname: Favia
authors.forename: Vito
authors.initials: V
authors.lastname: Racanelli
authors.forename: Gabriele
authors.initials: G
authors.lastname: Valentini
authors.forename: Federico
authors.initials: F
authors.lastname: Perosa
mesh_terms: D020816:Amino Acid Motifs; D000918:Antibody Specificity; D001323:Autoantibodies / Q000097:blood / Q000276:immunology* / Q000302:isolation & purification; D001324:Autoantigens / Q000737:chemistry* / Q000276:immunology*; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry* / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D012595:Scleroderma, Systemic / Q000097:blood / Q000276:immunology*
chemical_list: D001323:Autoantibodies; D001324:Autoantigens; C000619700:CENPA protein, human; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone; D007074:Immunoglobulin G