* Author for correspondence. Present address: Institut National de Recherche Agronomique, Laboratoire de Pathologie Infectieuse et mmunologie, 37380 Nouzilly, France. Tel. 47 42 78 67; fax 47 42 77 79
Summary: Twenty-two monoclonal antibodies (mAbs) specific for smooth lipopolysaccharide (S-LPS) were generated by fusion of spleen cells from mice infected with the rough Brucella melitensis strain B115 with the NSO myeloma. According to reactivity in enzyme-linked immunosorbent assay (ELISA) with O-polysaccharide (O-PS) and absence of reactivity with rough lipopolysaccharide (R-LPS), it was postulated that these mAbs recognized epitopes present on the O-PS. Most of the mAbs reacted equally well in ELISA and immunoblotting with S-LPS types of Brucella A and M dominant strains and were designated as specific for common (C) epitopes. Three mAbs were highly specific for M dominant S-LPS. All these mAbs, in contrast to a mAb specific for the A epitope, showed little or no cross-reactivity with Yersinia enterocolitica O:9 S-LPS. S-LPS of B. melitensis B115 was extracted and analysed by immunoblotting and ELISA with mAbs specific for A, M and C epitopes. Reactivity of the mAbs with this S-LPS was compared to reactivity with S-LPS of A and M dominant smooth Brucella strains. The results suggest that S-LPS of B. melitensis B115 bears mainly C epitopes and a few M epitopes. The very weak reactivity of this S-LPS with the mAb specific for the A epitope and the fact that the mAbs specific for C and M epitopes showed little or no cross-reactivity with Y. enterocolitica O:9 S-LPS suggest that O-PS from this rough strain could be used to distinguish Y. enterocolitica O:9 infection from Brucella infection. The potential value of the rough B. melitensis strain B115 as a vaccine strain is also discussed.
BundleD.R.,
CherwonogrodzkyJ.W.,
GidneyM.A.J.,
MeikleP.J.,
PerryM.B.,
PetersT.1989; Definition of Brucella A and M epitopes by monoclonal typing reagents and synthetic oligosaccharides.. Infection and Immunity 57:2829–2836
CloeckaertA.,
De WergifosseP.,
DubrayG.,
LimetJ.N.1990; Identification of seven surface-exposed Brucella outer membrane proteins by use of monoclonal antibodies: immunogold labeling for electron microscopy and enzyme-linked immunosorbent assay.. Infection and Immunity 58:3980–3987
CloeckaertA.,
JacquesI.,
De WergifosseP.,
DubrayG.,
LimetJ.N.1992a; 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.. Infection and Immunity 60:312–315
CloeckaertA.,
ZygmuntM.S.,
NicolleJ.-C.,
DubrayG.,
LimetJ.N.1992b; O-chain expression in the rough Brucella melitensis strain B115: induction of O-polysaccharide specific monoclonal antibodies and intracellular localization demonstrated byimmunoelectron microscopy.. Journal of General Microbiology 138:1211–1219
DouglasJ.T.,
PalmerD.A.1988; Use of monoclonal antibodies to identify the distribution of A and M epitopes on smooth Brucellaspecies.. Journal of Clinical Microbiology 26:1353–1356
Garin-BastujiB.,
BowdenR.A.,
DubrayG.,
LimetJ.N.1990; Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting analysis of smooth-lipopolysaccharide hetero-geneity among Brucella biovars related to A and M specificities.. Journal of Clinical Microbiology 28:2169–2174
HarmonB.G.,
AdamsL.G.,
FreyM.1988; Survival of rough and smooth strains of Brucella abortus in bovine mammary gland macrophages.. American Journal of Veterinary Research 49:1092–1097
JacquesI.,
Olivier-BernardinV.,
DubrayG.1991; Induction of antibody and protective responses in mice by Brucella O-polysaccharide-BSA conjugate.. Vaccine 9:896–900
JonesL.M.,
DiazR.,
TaylorA.G.1973; Characterization of allergens prepared from smooth and rough strains of Brucella melitensis.. British Journal of Experimental Pathology 54:492–508
KreutzerD.L.,
RobertsonD.C.1979; Surface macromolecules and virulence in intracellularparasitism: comparison of cell envelope components of smooth and rough strains of Brucella abortus.. Infection and Immunity 23:819–837
LeongD.,
DiazR.,
MilnerK.,
RudbachJ.,
WilsonJ.B.1970; Some structural and biological properties of Brucella endotoxin.. Infection and Immunity 1:174–182
LimetJ.N.,
BerbinschiA.,
CloeckaertA.,
CambiasoC.L.,
MassonP.L.1988; Longitudinal study of brucellosis in mice by immunoassay of lipopolysaccharide-related antigens in blood and urine.. Journal of Medical Microbiology 26:37–45
LimetJ.N.,
BosserayN.,
Garin-BastujiB.,
DubrayG.,
PlommetM.1989; Humoral immunity in mice mediated by monoclonal antibodies against the A and M antigens of Brucella.. Journal of Medical Microbiology 30:37–43
MeikleP.J.,
PerryM.B.,
CherwonogrodzkyJ.W.,
BundleD.R.1989; Fine structure of A and M antigens from Brucellabiovars.. Infection and Immunity 57:2820–2828
PalmerD.A.,
DouglasJ.T.1989; Analysis of Brucellalipopolysaccharide with specific and cross-reacting monoclonal antibodies.. Journal of Clinical Microbiology 27:2331–2337
RoopR.M.IIPreston-MooreD.,
BagchiT.,
SchurigG.G.1987; Rapid identification of smooth Brucella species with a monoclonal antibody.. Journal of Clinical Microbiology 25:2090–2093
TobiasL.,
SchurigG.G.,
CordesD.O.1992; Comparative behaviour of Brucella abortus strains 19 and RB51 in the pregnant mouse.. Research in Veterinary Science 53:179–183
VizcainoN.,
Fernandez-LagoL.1992; A rapid and sensitive method for the identification of Brucella species with a monoclonal antibody.. Research in Microbiology 143:513–518
ZygmuntM.S.,
DubrayG.,
BundleD.R.,
PerryM.B.1988; Purified native haptens of Brucella abortus B19 and B. melitensis16M reveal the lipopolysaccharide origin of the antigens.. Annales de l̓Institut Pasteur/Microbiologie 139:421–433