Brucella ovis, a naturally virulent rough Brucella species, is the aetiological agent of ram epididymitis. The identification of protective antigens is necessary to obtain a safe, specific subcellular vaccine. Monoclonal antibodies (MAbs) directed at both brucella outer-membrane proteins (OMPs) and rough lipopolysaccharide (R-LPS) in a mouse protection test were used to identify potential targets for humoral immunity. Mixtures of MAbs directed at the 16.5-, 25–27-, 31-34- and 36-38-kDa OMPs conferred significant protection 7 days after challenge with reference strain B. ovis 63/290 compared with controls receiving either saline or an anti-brucella O-polysaccharide MAb. Furthermore, an anti-R-LPS MAb tested alone conferred protection at a level comparable with that obtained with the mixture of anti-OMP MAbs. The combination of protective OMP MAbs with the anti-R-LPS MAb was also strongly protective. One combination of OMP MAbs, which bound intensely to B. ovis in vitro, was ineffective. These results indicate that B. ovis OMPs and R-LPS are targets for protective antibodies and that they can be regarded as candidates for ram epididymitis subcellular vaccines.
BlascoJ. M.,
MarinC. M.,
BarberánM.,
MoriyonI.,
DiazR. Immunization with Brucella melitensis Rev 1 against Brucella ovis infection of rams. Vet Microbiol1987; 14:381–392
ChinJ.,
TurnerB. Extraction of membrane antigens from Brucella ovis and an assessment of their serological activity by immunoblotting. J Gen Microbiol1990; 136:1615–1622
MyersD. M.,
SiniukA. A. Preliminary report on the development of a diffusion-in-gel method for the diagnosis of ram epididymitis. Appl Microbiol1970; 19:335–337
SuarezC. E.,
PachecoG. A.,
ViglioccoA. M. Immunochemical studies of oligosaccharides obtained from the lipopoly-saccharide of Brucella ovis. Vet Microbiol1990; 22:329–334
BlascoJ. M.,
GamazoC.,
WinterA. J. Evaluation of whole cell and subcellular vaccines against Brucella ovis in rams. Vet Immunol Immunopathol1993; 37:257–270
Jiménez de BaguésM. P.,
ElzerP. H.,
BlascoJ. M.,
MarinC. M.,
GamazoC.,
WinterA. J. Protective immunity to Brucella ovis in BALB/c mice following recovery from primary infection or immunization with subcellular vaccines. Infect Immun1994; 62:632–638
GamazoC.,
WinterA. J.,
MoriyonI.,
Riezu-BojJ. I.,
BlascoJ. M.,
DiazR. Comparative analyses of proteins extracted by hot saline or released spontaneously into outer-membrane blebs from field strains of Brucella ovis and Brucella melitensis. Infect Immun1989; 57:1419–1426
AltonG. G.,
JonesL. M.,
AngusR. D.,
VergerJ. M.Techniques for the brucellosis laboratory Institut National de la Recherche Agronomique; Paris, France: 1988
CloeckaertA.,
de WergifosseP.,
DubrayG.,
LimetJ. N. Identification of seven surface-exposed Brucella outer membrane proteins by use of monoclonal antibodies: immunogold labeling for electron microscopy and enzyme-linked immunosorbent assay. Infect Immun1990; 58:3980–3987
CloeckaertA.,
JacquesI.,
BosseraN. Protection conferred on mice by monoclonal antibodies directed against outer-membrane-protein antigens of Brucella. J Med Microbiol1991; 34:175–180
JacquesI.,
CloeckaertA.,
LimetJ. N.,
DubrayG. Protection conferred on mice by combinations of monoclonal antibodies directed against outer-membrane proteins or smooth lipopolysaccharide of Brucella. J Med Microbiol1992; 37:100–103
CloeckaertA.,
JacquesI.,
de WergifosseP.,
DubrayG.,
LimetJ. N. Protection against Brucella melitensis or Brucella abortus in mice with immunoglobulin G (IgG), IgA and IgM monoclonal antibodies specific for a common epitope shared by the Brucella A and M smooth lipopolysaccharides. Infect Immun1992; 60:312–315
LimetJ. N.,
PlommetA. M.,
DubrayG.,
PlommetM. Immunity conferred to mice by anti-LPS monoclonal antibodies in murine brucellosis. Ann Inst Pasteur/Immunol1987; 138:417–424
LimetJ. N.,
BosserayN.,
Garin-BastujiB.,
DubrayG.,
PlommetM. Humoral immunity in mice mediated by monoclonal antibodies directed against the A and M antigens of Brucella. J Med Microbiol1989; 30:37–43
CloeckaertA.,
JacquesI.,
BowdenR. A.,
DubrayG.,
LimetJ. N. Monoclonal antibodies to Brucella rough lipopolysaccharide: characterization and evaluation of their protective effect against B. abortus. Res Microbiol1993; 144:475–484
MontarazJ. A.,
WinterA. J.,
HunterD. M.,
SowaB. A.,
WuA. M.,
AdamsL. G. Protection against Brucella abortus in mice with O-polysaccharide-specific monoclonal antibodies. Infect Immun1986; 51:961–963
PhillipsM.,
DeyoeB. L.,
CanningP. C. Protection of mice against Brucella abortus infection by inoculation with monoclonal antibodies recognizing Brucella O-antigen. Am J Vet Res1989; 50:2158–2161