Immunological characterization of yersinia enterocolitica O:9 and O:3 LPS antigens by monoclonal antibodies

Weninger, Jürgen; Wartenberg, Klaus; Röllinghoff, Martin

doi:10.1016/s0176-6724(88)80173-1

Cited by 5 publications

(6 citation statements)

References 18 publications

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“…After centrifugation and washes, the bacteria were adjusted to a concentration of 109 bacteria per ml (optical density at 600 nm of 1.0). The expression of plasmidencoded proteins was controlled immunochemically in a dot blot assay, using monoclonal antibodies specific for YadA and YopE (28,29).…”

Section: Methodsmentioning

confidence: 99%

Mechanism of YadA-mediated serum resistance of Yersinia enterocolitica serotype O3

et al. 1992

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Complement activation via the alternative pathway was analyzed with isogenic strains of Yersinia enterocolitica serotype 03 differing in plasmid content (por p+ strains) or selective lack of YadA expression (YadAstrain). The p+ strain was serum resistant, even after antibody-enhanced complement activation. Serum sensitivity was observed with the p-and YadAstrains but was more pronounced in the p-strain. The p+ strain deposited less C5b-9(m) complexes on its surface than the p-and YadA-strains. No size difference, however, was detected with solubilized C5b-9(m) complexes obtained from resistant and sensitive strains. At the C3 level, it became evident that surface-bound C3b was degraded faster into iC3b on the p+ strain than on the pand YadAstrains. Our results demonstrate that YadA inhibits complement activation at the C3 and C9 level. As a result, reduced amounts of C5b-9(m) are generated on the surface of YadA-bearing bacteria. In addition, YadA seems to protect against the lytic action of those C5b-9(m) complexes whose deposition could not be prevented.

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Section: Methodsmentioning

confidence: 99%

Mechanism of YadA-mediated serum resistance of Yersinia enterocolitica serotype O3

et al. 1992

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“…Microagglutination assay. Heat-killed bacteria of the plasmid-free Y. enterocolitica serotype 09, strain Ye 09/6, and B. abortus S19 were used as antigens, as described in detail recently (23). Briefly, each well of a flat-bottom microculture plate (Dynatech, Plochingen, Federal Republic of Germany) received 108 heat-killed (100°C) bacteria in 50 ,ul of PBS and 50 ,lI of serum in serial dilutions, beginning with a dilution of 1:40.…”

Section: Methodsmentioning

confidence: 99%

“…Immunoblot analysis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was performed according to the method of Laemmli (15), and the immunoblot was set up according to a modification of the method of Towbin et al (21), as recently described (23). Briefly, 50 ,ul of a whole-cell bacterial suspension (OD, 1.0) or 100 ktg of the released proteins ofthe plasmid-carrying Ye 09/5 strain per 10 ,ul was heated with sample buffer (62.5 mM Tris hydrochloride, pH 6.8, containing 10% glycerol, 5% 2-mercaptoethanol, 2% sodium dodecyl sulfate, and 0.001% bromphenol blue) at 100°C for 5 min.…”

Section: Methodsmentioning

confidence: 99%

Differentiation of serological responses to Yersinia enterocolitica serotype O9 and Brucella species by immunoblot or enzyme-linked immunosorbent assay using whole bacteria and Yersinia outer membrane proteins

1990

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Serum samples from 134 patients showing by the microagglutination test serological cross-reactivity between Yersinia enterocolitica serotype 09 and Brucella spp. were analyzed by immunoblot and enzyme-linked immunosorbent assay techniques for the presence of antibodies directed against plasmid-encoded, yersiniaassociated outer membrane proteins (OMPs). Since these OMPs are exclusively expressed in pathogenic strains of Yersinia spp., this characteristic was chosen for serological differentiation of infections caused by these bacteria. The presence of antibodies against plasmid-encoded OMPs of pathogenic Yersinia spp. in patient sera appeared to be a suitable means to identify acute or recent infection with Y. enterocolitica serotype 09, whereas the failure to detect such antibodies indicated an acute or recent infection with Brucella spp. Serodiagnosis of acute and recent infections with Yersinia enterocolitica serotype 09 and Brucella spp. on the basis of

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“…Mab A6 reacted in enzyme immunoassay with purified 0-ag, recognized 100% of tested Y. enterocolitica 0:3 strains, and showed no cross-reactions with other bacteria (60). In addition to 0:3 also 0:9 0-ag specific Mab was described (88) Simple analysis of LPS is done by PAGE, either in the presence of SDS or DOC,30 which is usually combined with silver staining or immunoblotting. In PAGE analysis, LPS molecules are usually divided into two populations ( Fig.…”

Section: Bacteriophages and Other Tools For Lps Molecular Geneticsmentioning

confidence: 99%

Molecular genetics and biochemistry of Yersinia lipopolysaccharide

Skurnik

Zhang

1996

APMIS

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Studies on the molecular genetics of bacterial LPS1 serve at least two main purposes: (i) to help develop an understanding of the biology, biochemistry and genetics of this bacterial surface macro‐molecule, and (ii) to provide a basis for both vaccine development and virulence experiments. Both of these goals have been the driving force in studies of Yersinia LPS carried out during the last decade. Here we will review the progress made in the molecular genetics and biochemistry of Yersinia LPS. A deep understanding has been achieved with respect to Y. enterocolitica serotype 0:3, reaching as far as a detailed analysis of the gene clusters directing the biosynthesis of the outer core oligosaccharide and of the O‐ag2. The O‐ag gene clusters of Y. enterocolitica serotype 0:8 and Y. pseudotuberculosis serotypes 0:2a and 0:5a have also been cloned and partially characterized. LPS biosynthesis of these Yersinia species includes examples of the two major variations recognized in the biosynthesis of this macromolecule: (i) homopolymeric or O‐antigen polymerase‐independent biosynthesis, and (ii) heteropolymeric or O‐antigen polymerase‐dependent biosynthesis.

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Immunological characterization of yersinia enterocolitica O:9 and O:3 LPS antigens by monoclonal antibodies

Cited by 5 publications

References 18 publications

Mechanism of YadA-mediated serum resistance of Yersinia enterocolitica serotype O3

Mechanism of YadA-mediated serum resistance of Yersinia enterocolitica serotype O3

Differentiation of serological responses to Yersinia enterocolitica serotype O9 and Brucella species by immunoblot or enzyme-linked immunosorbent assay using whole bacteria and Yersinia outer membrane proteins

Molecular genetics and biochemistry of Yersinia lipopolysaccharide

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