title: Immunogenic HLA-DR-Presented Self-Peptides Identified Directly from Clinical Samples of Synovial Tissue, Synovial Fluid, or Peripheral Blood in Patients with Rheumatoid Arthritis or Lyme Arthritis.
abstract: Human leukocyte antigen-antigen D related (HLA-DR) molecules are highly expressed in synovial tissue (ST), the target of the immune response in chronic inflammatory forms of arthritis. Here, we used LC-MS/MS to identify HLA-DR-presented self-peptides in cells taken directly from clinical samples: ST, synovial fluid mononuclear cells (SFMC), or peripheral blood mononuclear cells (PBMC) from five patients with rheumatoid arthritis (RA) and eight with Lyme arthritis (LA). We identified 1593 non-redundant HLA-DR-presented peptides, derived from 870 source proteins. A total of 67% of the peptides identified in SFMC and 55% of those found in PBMC were found in ST, but analysis of SFMC/PBMC also revealed new antigen-presented peptides. Peptides were synthesized and examined for reactivity with the patients' PBMC. To date, three autoantigens in RA and four novel autoantigens in LA, presented in ST and/or PBMC, were shown to be targets of T- and B-cell responses in these diseases; ongoing analyses may add to this list. Thus, immunoprecipitation and LC-MS/MS can now identify hundreds of HLA-DR-presented self-peptides from individual patients' tissues or fluids with mixed cell populations. Importantly, identification of HLA-DR-presented peptides from SFMC or PBMC allows testing of more patients, including those early in the disease. Direct analysis of clinical samples facilitates identification of novel immunogenic T-cell epitopes.
authors.affiliation: Center for Biomedical Mass Spectrometry, Department of Biochemistry, Boston University School of Medicine , Boston, Massachusetts 02118, United States.
authors.forename: Qi
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Center for Immunology and Inflammatory Diseases, Division of Rheumatology, Allergy and Immunology, Massachusetts General Hospital, Harvard Medical School , Boston, Massachusetts 02114, United States.
authors.forename: Elise E
authors.initials: EE
authors.lastname: Drouin
authors.affiliation: Center for Biomedical Mass Spectrometry, Department of Biochemistry, Boston University School of Medicine , Boston, Massachusetts 02118, United States.
authors.forename: Chunxiang
authors.initials: C
authors.lastname: Yao
authors.affiliation: Center for Biomedical Mass Spectrometry, Department of Biochemistry, Boston University School of Medicine , Boston, Massachusetts 02118, United States.|National University of Defense Technology , Changsha, 410000 Hunan Province, China.
authors.forename: Jiyang
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Center for Biomedical Mass Spectrometry, Department of Biochemistry, Boston University School of Medicine , Boston, Massachusetts 02118, United States.
authors.forename: Yu
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Center for Biomedical Mass Spectrometry, Department of Biochemistry, Boston University School of Medicine , Boston, Massachusetts 02118, United States.
authors.forename: Deborah R
authors.initials: DR
authors.lastname: Leon
authors.affiliation: Center for Immunology and Inflammatory Diseases, Division of Rheumatology, Allergy and Immunology, Massachusetts General Hospital, Harvard Medical School , Boston, Massachusetts 02114, United States.
authors.forename: Allen C
authors.initials: AC
authors.lastname: Steere
authors.affiliation: Center for Biomedical Mass Spectrometry, Department of Biochemistry, Boston University School of Medicine , Boston, Massachusetts 02118, United States.
authors.forename: Catherine E
authors.initials: CE
authors.lastname: Costello
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D017951:Antigen Presentation; D001172:Arthritis, Rheumatoid / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D001324:Autoantigens / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology / Q000473:pathology; D015870:Gene Expression; D063990:Gene Ontology; D006684:HLA-DR Antigens / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D006801:Humans; D008193:Lyme Disease / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D008875:Middle Aged; D058977:Molecular Sequence Annotation; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology* / Q000302:isolation & purification; D013582:Synovial Fluid / Q000737:chemistry / Q000276:immunology*; D013583:Synovial Membrane / Q000737:chemistry / Q000276:immunology* / Q000473:pathology; D013601:T-Lymphocytes / Q000276:immunology / Q000473:pathology
chemical_list: D001324:Autoantigens; D006684:HLA-DR Antigens; D010455:Peptides
keywords: HLA-DR; Lyme arthritis; autoantigens; peripheral blood mononuclear cells; rheumatoid arthritis; synovial fluid mononuclear cells; synovial tissue