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Recombinant Human Monoclonal Autoantibodies Specific for Citrulline-Containing Peptides from Phage Display Libraries Derived from Patients with Rheumatoid Arthritis
JOS M.H. RAATS, EVELINE M. WIJNEN, GER J.M. PRUIJN, FRANK H.J. van den HOOGEN, and WALTHER J. van VENROOIJ
ABSTRACT.
Methods. Using lymphocytes from bone marrow or peripheral blood from RA patients, we constructed antibody fragment libraries representing the antibody repertoire of these individuals. Antibody fragments recognizing a citrulline-containing peptide were selected from these patient libraries. Individual antibody clones were analyzed for germline gene usage and reactivity toward citrullinated (auto)-antigens. Results. Sequence analysis of the cDNA encoding the 21 distinct antibody fragments that were obtained revealed a restricted germline gene usage. Individual antibody clones were positive in both antiperinuclear factor (APF) and antikeratin antibody (AKA) tests, stained citrullinated filaggrin and fibrinogen on Western blots, and reacted with subsets of citrulline-containing peptides in ELISA, but not with noncitrullinated peptides. Conclusion. Our report describes the first recombinant human Mab fragments reactive with citrulline-containing peptides. The restricted germline gene usage of these antibodies, and the fact that the VH alleles used are not present in all individuals, may indicate the existence of a genetic predisposition for the development of anticitrulline antibodies in individuals with these germline alleles. The selected antibody clones may facilitate studies on the role of these autoantibodies and their target antigens in the development of RA. (J Rheumatol 2003;30:1696-711) Key Indexing Terms:
RECOMBINANT ANTIBODY FRAGMENTS
From the Department of Biochemistry, University of Nijmegen; and the Department of Rheumatology, University Medical Centre St. Radboud, Nijmegen, The Netherlands. Supported by a grant from the Royal Netherlands Academy of Arts and Sciences. J.M.H. Raats, PhD; E.M. Wijnen, Ir.; G.J.M. Pruijn, PhD; W.J. van Venrooij, PhD, Department of Biochemistry, University of Nijmegen; F.H.J. van den Hoogen, MD, Department of Rheumatology, University Medical Centre St. Radboud. Address reprint requests to Dr. J.M.H. Raats, Department of Biochemistry 161, University of Nijmegen, PO Box 9101, NL-6500 HB Nijmegen, The Netherlands. E-mail: j.raats@ncmls.kun.nl Submitted June 18, 2002; revision accepted January 22, 2003. |