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Infection and Immunity, March 2000, p. 1542-1548, Vol. 68, No. 3
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Colonic Bacteria Express an Ulcerative Colitis pANCA-Related Protein Epitope

O. Cohavy,1 D. Bruckner,1 L. K. Gordon,2 R. Misra,3 B. Wei,1 M. E. Eggena,1 S. R. Targan,4,5 and J. Braun1,6,*

Departments of Pathology and Laboratory Medicine,1 Ophthalmology,2 and Medicine4 and Molecular Biology Institute,6 University of California, Los Angeles, California 90095; Department of Microbiology, Arizona State University, Tempe, Arizona 852873; and Inflammatory Bowel Disease Research Center, Cedars-Sinai Medical Center, Los Angeles, California 900485

Received 13 August 1999/Returned for modification 11 October 1999/Accepted 7 December 1999

Bacteria are a suspected pathogenic factor in inflammatory bowel disease, but the identity of the relevant microbial species remains unresolved. The pANCA autoantibody is associated with most cases of ulcerative colitis (UC) and hence reflects an immune response associated with the disease process. This study addresses the hypothesis that pANCA identifies an antigen(s) expressed by bacteria resident in the human colonic mucosa. Libraries of colonic bacteria were generated using aerobic and anaerobic microbiologic culture conditions, and bacterial pools and clonal isolates were evaluated for cross-reactive antigens by immunoblot analysis using the pANCA monoclonal antibody Fab 5-3. Two major species of proteins immunoreactive to pANCA monoclonal antibodies were detected in bacteria from the anaerobic libraries. Colony isolates of the expressing bacteria were identified as Bacteroides caccae and Escherichia coli. Isolation and partial sequencing of the B. caccae antigen identified a 100-kDa protein without database homologous sequences. The E. coli protein was biochemically and genetically identified as the outer membrane porin OmpC. Enzyme-linked immunosorbent assay with human sera demonstrated elevated immunoglobulin G anti-OmpC in UC patients compared to healthy controls. These findings demonstrate that a pANCA monoclonal antibody detects a recurrent protein epitope expressed by colonic bacteria and implicates colonic bacterial proteins as a target of the disease-associated immune response.

* Corresponding author. Mailing address: Department of Pathology and Lab Medicine, CHS 13-222, Los Angeles, CA 90095-1732. Phone: (310) 794-7953. Fax: (310) 825-5674. E-mail: jbraun{at}mednet.ucla.edu.

Infection and Immunity, March 2000, p. 1542-1548, Vol. 68, No. 3
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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