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Investigating the candidacy of lipopolysaccharide-based glycoconjugates as vaccines to combat Mannheimia haemolytica

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Abstract

Inner core lipopolysaccharide (LPS) has been shown to be conserved in the majority of veterinary strains from the species Mannheimia haemolytica, Actinobacillus pleuropneumoniae and Pasteurella multocida and as such is being considered as a possible vaccine antigen. The proof-in-principle that a LPS-based antigen could be considered as a vaccine candidate has been demonstrated from studies with monoclonal antibodies raised to the inner core LPS of Mannheimia haemolytica, which were shown to be both bactericidal and protective in a mouse model of disease. In this study we confirm and extend the candidacy of the inner core LPS by demonstrating that it is possible to elicit functional antibodies against Mannheimia haemolytica wild-type strains following immunisation of rabbits with glycoconjugates elaborating the conserved inner core LPS antigen. The present study describes a conjugation strategy that uses amidases produced by Dictyostelium discoideum, targeting the amino functionality created by the amidase activity as the attachment point on the LPS molecule. To protect the amino functionality on the phosphoethanolamine (PEtn) residue of the inner core, we developed a novel blocking and unblocking strategy with t-butyl oxycarbonyl. A maleimide-thiol linker strategy with the thiol linker on the carboxyl residues of the carrier protein and the maleimide linker on the carbohydrate resulted in a high loading of carbohydrates per carrier protein. Immunisation derived antisera from rabbits recognised fully extended Mannheimia haemolytica LPS and whole cells from serotypes 1 and 2, despite a somewhat immunodominant response to the linkers also being observed. Moreover, bactericidal activity was demonstrated to a strain elaborating the immunising carbohydrate antigen and crucially to wild-type cells of serotypes 1 and 2, thus further supporting the consideration of inner core LPS as a potential vaccine antigen to combat disease caused by Mannheimia haemolytica.

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Acknowledgements

We thank Perry Fleming (core Bacterial Culture Facility) for large scale biomass production and Jacek Stupak for mass spectrometry. We also thank Drs. Srikumaran Subramaniam and William Foreyt, Washington State University and Anthony Confer, Oklahoma State University, for providing us with M. haemolytica serotype A2 isolates from domestic sheep and cattle, respectively. We thank Annie Aubry and Dr. Susan Logan for helpful discussions. We are grateful to Novartis Vaccines for providing us with CRM197.

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  1. Institute for Biological Sciences, National Research Council, 100, Sussex Drive, Ottawa, ON, K1A 0R6, Canada

    Frank St. Michael, Chantelle Cairns, Amy Lea Filion, Dhamodharan Neelamegan, Suzanne Lacelle & Andrew D. Cox

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  1. Frank St. Michael
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  2. Chantelle Cairns
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Correspondence to Andrew D. Cox.

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St. Michael, F., Cairns, C., Filion, A.L. et al. Investigating the candidacy of lipopolysaccharide-based glycoconjugates as vaccines to combat Mannheimia haemolytica . Glycoconj J 28, 397–410 (2011). https://doi.org/10.1007/s10719-011-9339-0

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