Detection of Multiple Autoantibodies in Patients with Ankylosing Spondylitis Using Nucleic Acid Programmable Protein Arrays

Wright, Cynthia; Sibani, Sahar; Trudgian, David; Fischer, Roman; Kessler, Benedikt; LaBaer, Joshua; Bowness, Paul

doi:10.1074/mcp.M9.00384

Detection of Multiple Autoantibodies in Patients with Ankylosing Spondylitis Using Nucleic Acid Programmable Protein Arrays*

From the ‡Human Immunology Unit, Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, University of Oxford, Oxford OX3 9DS, United Kingdom,
the **Harvard Institute of Proteomics, Harvard Medical School, Cambridge, Massachusetts 02141,
the §Central Proteomics Facility, Henry Wellcome Building for Molecular Physiology, Department of Clinical Medicine, University of Oxford, Oxford OX3 7BN, United Kingdom,
the ‡‡Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-5001, and
the §§Nuffield Orthopaedic Centre, National Health Service Trust, Headington, Oxford OX3 7LD, United Kingdom

‖ To whom correspondence should be addressed. Tel.: 44-1865-287804; Fax: 44-1865-287787; E-mail: cwdrakesmith{at}gmai1.com and paul.bowness{at}imm.ox.ac.uk.

↵¶ These authors contributed equally to this work.

Abstract

Ankylosing spondylitis (AS) is a common, inflammatory rheumatic disease that primarily affects the axial skeleton and is associated with sacroiliitis, uveitis, and enthesitis. Unlike other autoimmune rheumatic diseases, such as rheumatoid arthritis or systemic lupus erythematosus, autoantibodies have not yet been reported to be a feature of AS. We therefore wished to determine whether plasma from patients with AS contained autoantibodies and, if so, characterize and quantify this response in comparison to patients with rheumatoid arthritis (RA) and healthy controls. Two high density nucleic acid programmable protein arrays expressing a total of 3498 proteins were screened with plasma from 25 patients with AS, 17 with RA, and 25 healthy controls. Autoantigens identified were subjected to Ingenuity Pathway Analysis to determine the patterns of signaling cascades or tissue origin. 44% of patients with ankylosing spondylitis demonstrated a broad autoantibody response, as compared with 33% of patients with RA and only 8% of healthy controls. Individuals with AS demonstrated autoantibody responses to shared autoantigens, and 60% of autoantigens identified in the AS cohort were restricted to that group. The autoantibody responses in the AS patients were targeted toward connective, skeletal, and muscular tissue, unlike those of RA patients or healthy controls. Thus, patients with AS show evidence of systemic humoral autoimmunity and multispecific autoantibody production. Nucleic acid programmable protein arrays constitute a powerful tool to study autoimmune diseases.

Footnotes

↵* This work was supported by funding provided by the National Institutes of Health Research Biomedical Research Centre Oxford, the Medical Research Council, Action Medical Research (UK), National Science and Engineering Research Council of Canada, and the European Molecular Biology Organization. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵ This article contains supplemental material.