Putative β-Barrel Outer Membrane Proteins of the Bovine Digital Dermatitis-Associated Treponemes: Identification, Functional Characterization, and Immunogenicity

Staton, GJ ORCID: 0000-0003-4100-8597, Carter, SD ORCID: 0000-0002-3585-9400, Ainsworth, S, Mullin, J, Smith, RF ORCID: 0000-0003-0944-310X and Evans, NJ ORCID: 0000-0002-2950-1007 (2020) Putative β-Barrel Outer Membrane Proteins of the Bovine Digital Dermatitis-Associated Treponemes: Identification, Functional Characterization, and Immunogenicity. INFECTION AND IMMUNITY, 88 (5). e00050-e00020.

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Official URL: http://dx.doi.org/10.1128/iai.00050-20

Abstract

Bovine digital dermatitis (BDD), an infectious disease of the bovine foot with a predominant treponemal etiology, is a leading cause of lameness in dairy and beef herds worldwide. BDD is poorly responsive to antimicrobial therapy and exhibits a relapsing clinical course; an effective vaccine is therefore urgently sought. Using a reverse vaccinology approach, the present study surveyed the genomes of the three BDD-associated Treponema phylogroups for putative β-barrel outer membrane proteins and considered their potential as vaccine candidates. Selection criteria included the presence of a signal peptidase I cleavage site, a predicted β-barrel fold, and cross-phylogroup homology. Four candidate genes were overexpressed in Escherichia coli BL21(DE3), refolded, and purified. Consistent with their classification as β-barrel OMPs, circular-dichroism spectroscopy revealed the adoption of a predominantly β-sheet secondary structure. These recombinant proteins, when screened for their ability to adhere to immobilized extracellular matrix (ECM) components, exhibited a diverse range of ligand specificities. All four proteins specifically and dose dependently adhered to bovine fibrinogen. One recombinant protein was identified as a candidate diagnostic antigen (disease specificity, 75%). Finally, when adjuvanted with aluminum hydroxide and administered to BDD-naive calves using a prime-boost vaccination protocol, these proteins were immunogenic, eliciting specific IgG antibodies. In summary, we present the description of four putative treponemal β-barrel OMPs that exhibit the characteristics of multispecific adhesins. The observed interactions with fibrinogen may be critical to host colonization and it is hypothesized that vaccination-induced antibody blockade of these interactions will impede treponemal virulence and thus be of therapeutic value.

Item Type:	Article
Uncontrolled Keywords:	Treponema, bovine digital dermatitis, outer membrane proteins, reverse vaccinology
Depositing User:	Symplectic Admin
Date Deposited:	13 Mar 2020 16:40
Last Modified:	18 Jan 2023 23:57
DOI:	10.1128/IAI.00050-20
Open Access URL:	https://doi.org/10.1128/IAI.00050-20
Related URLs:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3078936