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Journal of Clinical Microbiology, December 2000, p. 4412-4419, Vol. 38, No. 12
Center for Molecular Medicine and Infectious
Diseases, Virginia-Maryland Regional College of Veterinary Medicine,
Virginia Polytechnic Institute and State University, Blacksburg,
Virginia 240611; Institute for
Biological Sciences, National Research Council of Canada, Ottawa,
Ontario K1A 0R6, Canada2; and
Departments of Pediatrics and Microbiology, University of
Pennsylvania School of Medicine, Philadelphia, Pennsylvania
191043
Received 18 May 2000/Returned for modification 29 August
2000/Accepted 24 September 2000
The lipooligosaccharide (LOS) of Haemophilus somnus
undergoes antigenic phase variation, which may facilitate evasion from the bovine host immune response and/or colonization and dissemination. However, LOS antigenic diversity in H. somnus has not been
adequately investigated. In this study, monoclonal antibodies (MAbs)
specific to various LOS epitopes were used to investigate antigenic
variation and stability in LOS from H. somnus strains and
phase variants. Clinical isolates of H. somnus exhibited
intrastrain, as well as interstrain, antigenic heterogeneity in LOS
when probed with MAbs to outer core oligosaccharide epitopes in an
enzyme-linked immunosorbent assay (ELISA). However, epitopes
reactive with MAbs directed predominately to the inner core heptose
region were highly conserved. At least one epitope, which was
expressed in few strains, was identified. One LOS component affected by
phase variation was identified as phosphorylcholine (PCho), which is
linked to the primary glucose residue. Inhibition ELISA,
immunoblotting, and electrospray-mass spectrometry were used to confirm
that MAb 5F5.9 recognized PCho. LOS reactivity with MAb 5F5.9 was
associated with loss of most of the outer core oligosaccharide,
indicating that reactivity with PCho was affected by phase variation of
the glycose residues in this region. Our results indicate that outer core epitopes of H. somnus LOS exhibit a high degree of
random, phase-variable antigenic heterogeneity and that such
heterogeneity must be considered in the design of vaccines and
diagnostic tests.
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Antigenic Diversity of Haemophilus
somnus Lipooligosaccharide: Phase-Variable Accessibility of the
Phosphorylcholine Epitope
*
Corresponding author. Mailing address: College of
Veterinary Medicine, Virginia Tech, 1410 Prices Fork Rd., CMMID,
Blacksburg, VA 24061. Phone: (540) 231-4692. Fax: (540) 231-3426. E-mail: tinzana{at}vt.edu.
Journal of Clinical Microbiology, December 2000, p. 4412-4419, Vol. 38, No. 12
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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