Abstract

Western blot analysis of the human antibody response to Campylobacter jejuni cellular antigens during gastrointestinal infection. I Nachamkin and A M Hart ABSTRACT Western blot analysis was used to identify antigenic components of Campylobacter jejuni whole cells and outer membranes that elicit antibody responses in patients with campylobacter enteritis. Acute- and convalescent-phase sera from eight patients were analyzed for antibody activity against their homologous infecting strains and heterologous clinical isolates. Whole-cell and Sarkosyl-insoluble membrane components were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred electrophoretically to nitrocellulose paper for immunoblotting experiments. After the separated components were probed with patient sera, antibody binding was detected by autoradiography with 125I-protein A. Using this method, we detected several immunogenic components in whole cells and outer membranes. In the acute-phase response of some patients to infection, two to three components with approximate molecular weights of 66,000 (p66), 43,000 to 46,000 (major outer membrane protein), and 12,000 (p12) were detected in immunoblots. Convalescent-phase sera showed a more broad array of antibody binding to cell components. p66, shown to be campylobacter flagellin, was the major immunodominant component in almost all sera tested, however, p66 was not a major protein in Coomassie blue-stained gels. The major outer-membrane protein also bound to antibody, but with less intensity than p66. In general, the antibody specificity of patient sera was not limited to the homologous infecting strain, and antibodies cross-reacted with most components in heterologous strains. A low-molecular-weight component, identified as lipopolysaccharide with a modified silver stain, showed serological specificity for some patient sera. The results of this study showed that the antibody response of patients with campylobacter enteritis to C. jejuni antigens is variable. Flagellin appeared to be the major immunodominant component during infection. CiteULike Connotea Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter What's this? « Previous | Next Article » Table of Contents This Article J. Clin. Microbiol. January 1985 vol. 21 no. 1 33-38 » Abstract PDF Classifications Research Article Services Email this article to a colleague Similar articles in ASM journals Alert me when this article is cited Alert me if a correction is posted Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of JCM Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Load citing article information Citing articles via Web of Science Citing articles via Google Scholar Google Scholar Articles by Nachamkin, I. Articles by Hart, A. M. Search for related content PubMed PubMed citation Articles by Nachamkin, I. Articles by Hart, A. M. Related Content Load related web page information Social Bookmarking CiteULike Connotea Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter What's this? current issue December 2011, volume 49, issue 12 Alert me to new issues of JCM About JCM Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy JCM RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0095-1137 Online ISSN: 1098-660X Copyright © 2011 by the American Society for Microbiology. For an alternate route to JCM .asm.org, visit: http://intl- JCM .asm.org | More Info» var gaJsHost = (("https:" == document.location.protocol) ? "https://ssl." : "http://www."); document.write(unescape("%3Cscript src='" + gaJsHost + "google-analytics.com/ga.js' type='text/javascript'%3E%3C/script%3E")); var pageTracker = _gat._getTracker("UA-5821458-10"); pageTracker._trackPageview();