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Infection and Immunity, June 2000, p. 3469-3474, Vol. 68, No. 6
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Characterization of Hemolysin of Moraxella bovis Using a Hemolysis-Neutralizing Monoclonal Antibody

F. Mark Billson,1,dagger Colin Harbour,2 Wojtek P. Michalski,3 Jan M. Tennent,3 John R. Egerton,1 and Jennifer L. Hodgson1,*

Department of Veterinary Clinical Sciences, University of Sydney, Camden, New South Wales 2570,1 Department of Infectious Diseases, University of Sydney, Sydney 2000,2 and CSIRO, Division of Animal Health, Australian Animal Health Laboratory, Geelong, Victoria 3220,3 Australia

Received 23 November 1999/Returned for modification 17 January 2000/Accepted 20 March 2000

A concentrated bacterial culture supernatant from the hemolytic Moraxella bovis strain UQV 148NF was used to immunize mice and generate monoclonal antibodies (MAbs). One, MAb G3/D7, neutralized the hemolytic activity of M. bovis and recognized a 94-kDa protein by Western blot analysis in hemolytic M. bovis strains representing each of the different fimbrial serogroups. Exposure of corneal epithelial cells to M. bovis concentrated culture supernatants demonstrated a role for an exotoxin in the pathogenesis of infectious bovine keratoconjunctivitis, while neutralization of hemolytic and cytotoxic activities by MAb G3/D7 implies that these activities are related or have common epitopes. The action of M. bovis hemolysin was further characterized in sheep erythrocyte preparations with a binding step and Ca2+ required for lysis to proceed, similar to the RTX family of bacterial exotoxins. Neutralization of lytic activity in vitro is evidence for the presence of M. bovis antigens, which may be capable of protecting cattle from the development of infectious bovine keratoconjunctivitis.

* Corresponding author. Mailing address: University Veterinary Centre, Camden PMB 4, Narellan NSW 2567, Australia. Phone: 612-4655 0745. Fax: 612-4655 6942. E-mail: jennih{at}camden.usyd.edu.au.

dagger Present address: Department of Small Animal Clinical Studies, University of Glasgow, Glasgow, United Kingdom.

Infection and Immunity, June 2000, p. 3469-3474, Vol. 68, No. 6
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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