Composition and antigenic activity of the oligosaccharide moiety of Haemophilus influenzae type b lipooligosaccharide

Inzana, Thomas J.; Seifert, Wayne; Williams, R. R.

doi:10.1128/iai.48.2.324-330.1985

Cited by 55 publications

(27 citation statements)

References 46 publications

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“…The CP was purified as previously describe (10). LPS was extracted with hot 45% phenol and purified by ultracentrifugation as previously described (11,35).…”

Section: Methodsmentioning

confidence: 99%

Serotype specificity and immunogenicity of the capsular polymer of Haemophilus pleuropneumoniae serotype 5

Inzana¹,

Mathison²

1987

Infect Immun

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Serotyping of Haemophilus pleuropneumoniae and serologic assays for detection of serotype-specific antibody are problematic due to the potential cross-reactivity of the crude antigens used for raising immune serum or for serology. The capsular polymer (CP) of H. pleuropneumoniae serotype 5 was investigated for serotypespecific activity with antiserum to whole cells or with antiserum made monospecific to CP by adsorption with a capsule-deficient mutant. When antiserum to whole cells or monospecific antiserum to CP was tested against purified CP from serotypes 1 to 7 by immunodiffusion or enzyme-linked immunosorbent assay, only capsules of serotype 5 were reactive. In addition, only encapsulated serotype 5 cells reacted with serum monospecific to CP in an indirect immunofluorescent-antibody assay. Serotype-specific antibody was completely inhibited in each assay by preincubation of purified CP with the serum. Antiserum to whole cells of H. pleuropneumoniae serotype 5 contained antibodies to proteins and lipopolysaccharide; these antibodies cross-reacted with antigens of heterologous serotypes by dot-blot enzyme-linked immunosorbent assay and immunoblotting. The antigenic activity of CP was stable after heating for at least 30 min at 100°C. High titers of antibody to CP were present in the sera of rabbits immunized intravenously with whole log-phase cells or in the convalescent sera of pigs experimentally infected with H. pleuropneumoniae serotype 5. However, the purified CP was poorly immunogenic in rabbits and swine. Our results indicate that the capsule is the serotype-specific antigen of H. pleuropneumoniae and that a monospecific antiserum to capsule or purified capsule should be used for serotyping or serologic assays, respectively.

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“…The CP was purified as previously describe (10). LPS was extracted with hot 45% phenol and purified by ultracentrifugation as previously described (11,35).…”

Section: Methodsmentioning

confidence: 99%

Serotype specificity and immunogenicity of the capsular polymer of Haemophilus pleuropneumoniae serotype 5

Inzana¹,

Mathison²

1987

Infect Immun

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“…Relatively little is known about the physical structure of Hib LOS. Biochemical analysis has indicated that the oligosaccharide moiety of Hib LOS contains several different (20,30,45). The LOS of Hib also appears to be relatively high in phosphate content (45).…”

Section: Discussionmentioning

confidence: 99%

Antigenic evidence for simultaneous expression of two different lipooligosaccharides by some strains of Haemophilus influenzae type b

et al. 1989

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Isolates of Haemophilus influenzae type b (Hib) can be divided into three antigenic groups based on their reactivities with a set of two monoclonal antibodies (MAbs) directed against epitopes in the oligosaccharide region of Hib lipooligosaccharide (LOS) (P. A. Gulig, C. C. ratrick, L. Hermanstorfer, G. H. McCracken, Jr., and E. J. Hansen, Infect. Immun. 55:513-520, 1987). Approximately 24% of Hib strains react with both of these LOS-specific MAbs. Immunoprecipitation experiments involving several of these strains indicated that the epitopes recognized by these MAbs resided in two different LOS molecules, both of which were synthesized by these particular Hib strains. In addition, Western blot (immunoblot) analysis of proteinase K-treated cell extracts of these strains that had been subjected to sodium dodecyl sulfate-polyacrylamide gradient gel electrophoresis revealed two different LOS staining patterns when they were probed independently with the two MAbs. Colony blot radioimmunoassay of hundreds of colonies of one of these Hib strains showed that each colony bound both MAbs. Immune electron microscopy confirmed that individual cells of this same Hib strain expressed both types of LOS molecule at the same time. An antibody accessibility radioimmunoassay was used to show that different Hib strains of this type varied in the relative amounts of each of the two MAbs that they could bind to their cell surfaces. These findings indicate that some Hib strains can synthesize two antigenically distinct LOS molecules simultaneously.

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“…Complexes of lipopolysaccharide to polymyxin-B were also prepared (4,48). The lipid A fractions from the monkey isolate of P. gingivalis, B. forsythus (strain #9610) and A. actinomycetemcomitans (strain Y4) were prepared by mild acetic acid hydrolysis of hpopolysaccharide, centrifugation and chloroform extraction (15,24). The fatty acid and sugar composition of lipopolysaccharide and lipid A preparations were determined by gas-liquid chromatography with a Hewlett-Packard #5890 system equipped with an HP-1 capillary column by the method of Bryn & Jantzen (6) as described previously (41).…”

Section: Antigen Preparationsmentioning

confidence: 99%

Shared antigens of Porphymmonas gingivalis and Bacteroides forsythus

Vasel

Sims

Bainbridge

et al. 1996

Oral Microbiology and Immunology

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Periodontitis in humans is caused by a group of predominantly gram-negative, anaerobic bacteria among which Porphyromonas gingivalis and Bacteroides forsythus are prominent. A similar group is present and presumably plays a similar role in experimental periodontitis in the primate Macaca fascicularis. Nevertheless, immunization using a vaccine containing only killed P. gingivalis suppresses the progress of experimental periodontitis in M. fascicularis. We investigated the hypothesis that gram-negative periodontopathic bacterial may share antigens, and immunization with one species may induce antibodies reactive with other gram-negative species. Using enzyme-linked immunosorbent assay (ELISA), Western and dot immunoblots with nonabsorbed and absorbed and immune and preimmune sera we show that monkeys immunized with P. gingivalis produce antibodies reactive not only with antigens of P. gingivalis but also with those of B. forsythus. Similarly, rabbits immunized with P. gingivalis or with B. forsythus produce antibodies that react with antigens of both bacteria. Cross-reactive antibodies bind to epitopes in lipid A and possibly in core carbohydrate of lipopolysaccharide. Using complexes of lipopolysaccharide with polymyxin B, bovine serum albumin and apolipoprotein A1 specificity of binding was documented. Using sera from monkeys immunized with P. gingivalis, cross-reactivity with Actinobacillus actinomycetemcomitans could not be demonstrated by ELI-SA, although binding to lipopolysaccharide but not to lipid A was demonstrated by Western and dot immunoblots. Antibodies to shared lipopolysaccharide epitopes of periodontopathic bacteria may account, at least in part, for the immune protection observed in immunized monkeys, and shared epitopes may have potential as a vaccine for periodontitis in humans.

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Composition and antigenic activity of the oligosaccharide moiety of Haemophilus influenzae type b lipooligosaccharide

Cited by 55 publications

References 46 publications

Serotype specificity and immunogenicity of the capsular polymer of Haemophilus pleuropneumoniae serotype 5

Serotype specificity and immunogenicity of the capsular polymer of Haemophilus pleuropneumoniae serotype 5

Antigenic evidence for simultaneous expression of two different lipooligosaccharides by some strains of Haemophilus influenzae type b

Shared antigens of Porphymmonas gingivalis and Bacteroides forsythus

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