Mutants in the lipopolysaccharide of Brucella ovis are attenuated and protect against B. ovis infection in mice
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Brucella spp. are Gram-negative bacteria that behave as facultative intracellular parasites of a variety of mammals. This genus includes smooth (S) and rough (R) species that carry S and R lipopolysaccharides (LPS), respectively. S-LPS is a virulence factor, and mutants affected in the S-LPS O-polysaccharide (R mutants), core oligosaccharide or both show attenuation. However, B. ovis is naturally ... [++]
Brucella spp. are Gram-negative bacteria that behave as facultative intracellular parasites of a variety of mammals. This genus includes smooth (S) and rough (R) species that carry S and R lipopolysaccharides (LPS), respectively. S-LPS is a virulence factor, and mutants affected in the S-LPS O-polysaccharide (R mutants), core oligosaccharide or both show attenuation. However, B. ovis is naturally R and is virulent in sheep. We studied the role of B. ovis LPS in virulence by mutating the orthologues of wadA, wadB and wadC, three genes known to encode LPS core glycosyltransferases in S brucellae. When mapped with antibodies to outer membrane proteins (Omps) and R-LPS, wadB and wadC mutants displayed defects in LPS structure and outer membrane topology but inactivation of wadA had little or no effect. Consistent with these observations, the wadB and wadC but not the wadA mutants were attenuated in mice. When tested as vaccines, the wadB and wadC mutants protected mice against B. ovis challenge. The results demonstrate that the LPS core is a structure essential for survival in vivo not only of S brucellae but also of a naturally R Brucella pathogenic species, and they confirm our previous hypothesis that the Brucella LPS core is a target for vaccine development. Since vaccine B. melitensis Rev 1 is S and thus interferes in serological testing for S brucellae, wadB mutant represents a candidate vaccine to be evaluated against B. ovis infection of sheep suitable for areas free of B. melitensis. [--]
Linked immunosorbent assay, Outer membrane protein, Monoclonal-antibodies, O-polysaccharide, Rough lipopolysaccharide, OMP25/OMP31 family, DNA polymorphism, Host-range, Melitensis, Vaccines
Veterinary Research 2014, 45:72
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Universidad Pública de Navarra. Instituto de Agrobiotecnología / Nafarroako Unibertsitate Publikoa. Agrobioteknologiako Institutua
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This work was funded by Ministerio de Economía y Competitividad of Spain (reference project AGL2011-30453-C04) and Fundación para la Investigación Médica Aplicada (FIMA). Financial support to PS-L from Ministerio de Economía y Competitividad of Spain (reference BES-2009-015656), AZ-B from Universidad Pública de Navarra, and BSR from CSIC and European Social Fund (Programa JAE-Doc) are also gratefully acknowledged.
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