The Surface Properties of Neisseria gonorrhoeae: Isolation of the Major Components of the Outer Membrane

Heckels, John E.

doi:10.1099/00221287-99-2-333

Cited by 106 publications

(95 citation statements)

References 24 publications

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“…mAbs SM20, SM21, SM22 and SM24, which react with PIB from a number of gonococcal strains, were obtained following sequential immunization of mice with outer membranes from a variety of strains as described in detail previously . mAbs SM198 and SM203 were obtained subsequently using similar protocols after immunization of mice with outer membranes of strain P9 which had first been extracted with sodium cholate (Heckels, 1977).…”

Section: Methodsmentioning

confidence: 99%

“…Despite antigenic differences between the strains R10 and MS11, the inferred amino acid sequences show extensive homology, suggesting that limited structural variations may be responsible for generating antigenic diversity. In this paper we report the inferred amino acid sequence of PIB from strain P9, the immunobiology of which has been extensively studied (Heckels, 1977;Lambden & Heckels, 1979;Virji et al, 1986;Heckels et al, 1989Heckels et al, , 1990, and demonstrate that sequence variations occur between a number of other serovars in two discrete regions of the PIB molecule. In addition, we have used the sequence information to construct synthetic peptides and map the epitopes recognized by both the typespecific and cross-reacting, protective, anti-PIB mAbs.…”

Section: And Othersmentioning

confidence: 99%

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Gonococcal outer-membrane protein PIB: comparative sequence analysis and localization of epitopes which are recognized by type-specific and cross-reacting monoclonal antibodies

Butt

Virji

Vayreda

et al. 1990

Journal of General Microbiology

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Comparison of the inferred amino acid sequence of outer-membrane protein PIB from gonococcal strain P9 with those from other serovars reveals that sequence variations occur in two discrete regions of the molecule centred on residues 1% (Varl) and 237 (Var2). A series of peptides spanning the amino acid sequence of the protein were synthesized on solid-phase supports and reacted with a panel of monoclonal antibodies (mAbs) which recognize either type-specific or conserved antigenic determinants on PIB. Four type-specific mAbs reacted with overlapping peptides in Varl between residues 192-198. Analysis of the effect of amino acid substitutions revealed that the mAb specificity is generated by differences in the effect of single amino acid changes on mAb binding, so that antigenic differences between strains are revealed by different patterns of reactivity within a panel of antibodies. The variable epitopes in Varl recognized by the type-specific mAbs lie in a hydrophilic region of the protein exposed on the gonococcal surface, and are accessible to complement-mediated bactericidal lysis. In contrast, the epitope recognized by mAb SM198 is highly conserved but is not exposed in the native protein and the antibody is non-bactericidal. However, the conserved epitope recognized by mAb SM24 is centred on residues 198-199, close to Varl, and is exposed for bactericidal killing.

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Section: Methodsmentioning

confidence: 99%

Section: And Othersmentioning

confidence: 99%

Gonococcal outer-membrane protein PIB: comparative sequence analysis and localization of epitopes which are recognized by type-specific and cross-reacting monoclonal antibodies

Butt

Virji

Vayreda

et al. 1990

Journal of General Microbiology

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“…Previous work in this laboratory has involved the isolation of individual gonococcal surface antigens in a pure form as a prelude to biological studies. Thus lipopolysaccharide (Stead et al, 1975), pili (Robertson, Vincent & Ward, 1977) and two major outer membrane proteins (Heckels, 1977) have been purified from Neisseria gonorrhoeae ~9 .…”

Section: Introductionmentioning

confidence: 99%

The Isolation of a New Outer Membrane Protein from the Parent Strain of Neisseria gonorrhoeae P 9

Heckels

Everson

1978

Journal of General Microbiology

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“…The dose of antigen from strain P9(PS) for each injection was for one rabbit 50 pg of pili (Robertson et al, 1977); for the second 100 pg of colony type T4 outer membrane; for the third 50 pg of protein I (m.w. 36 500 daltons) extracted from the outer membrane (Heckels, 1977); for the fourth rabbit 50 pg of protein I1 (m.w. 24000 daltons) extracted from the outer membrane; and for the fifth 50 pg of lipopolysaccharide (Stead et al, 1975).…”

mentioning

confidence: 99%

“…Strain P9 (PS) gonococci were grown in bulk on G C Agar Base (Difco) and the pili were purified from them (Robertson, Vincent and Ward, 1977). Outer envelope was prepared from the residual depilated organisms by lithium acetate extraction and the two major outer envelope proteins were purified (Heckels, 1977). Gonococcal lipopolysaccharide (LPS) was extracted by the phenol-water method and further purified by enzyme digestion as described by Stead et al, (1975).…”

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confidence: 99%

Protection by Monospecific Gonococcal Antisera of the Chicken Embryo Challenged With Neisseria Gonorrhoeae

Robertson

1979

Journal of Medical Microbiology

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RECENT studies with the chick embryo as a model have investigated the infecting ability of virulent and non-virulent colony types of gonococci (Kellogg et al., 1963) and the effect of the inoculation route (Buchanan and Gotschlich, 1973;Bumgarner and Finkelstein, 1973;Foster and Vinson, 1977), but little is known about the pathogenesis of this infection.To initiate infection after intravenous challenge, gonococci must be resistant to the embryo's phagocytic defence systems and be capable of tissue invasion. Surface components of the gonococcus are presumably responsible and for this reason we have raised antisera to single purified components of the gonococcal surface and investigated the role of these sera in the protection of the chick embryo against gonococcal infection. MATERIALS AND METHODSGonococcus strain. Neisseria gonorrhoeae strain P9(PS) was isolated from a patient with gonorrhoea and was freeze-dried after minimum subculture. Colony types TI, T2 and T4 (Kellogg et al., 1963) were selected and stored in liquid nitrogen (Ward and Watt, 1971). The gonococci were grown overnight on solid gonococcal (GC) medium at 36°C in an atmosphere of air with 5% carbon dioxide and at a relative humidity of 100%. A standard inoculum of gonococci was prepared from a suspension containing approximately 1 x 1 O8 organisms per ml in organ culture medium (OCM) followed by centrifugation for 5 min. at 1500 g to remove any large clusters of organisms. The number of live gonococci in the inoculum was determined by a viable-count procedure.Media. Solid G C medium was prepared from GC Base (Difco) with the addition of 1% of Agar No. 1 (Oxoid) and 2% of Kellog supplement (Kellog et al., 1963) modified to contain 0.2% ferric citrate. The organ-culture medium (Gibco Biocult, Paisley, Scotland) was Basal Medium Eagle with Hanks salts and Hepes buffer to which was added glutamine to give a final concentration of 2 m~, and the concentration of Hepes buffer was raised to 5 0 m~. Preparation ofantigens. Strain P9 (PS) gonococci were grown in bulk on G C Agar Base (Difco) and the pili were purified from them (Robertson, Vincent and Ward, 1977). Outer envelope was prepared from the residual depilated organisms by lithium acetate extraction and the two major outer envelope proteins were purified (Heckels, 1977). Gonococcal lipopolysaccharide (LPS) was extracted by the phenol-water method and further purified by enzyme digestion as described by Stead et al., (1975).Antisera. Adult New Zealand white rabbits were given a course of five subcutaneous injections each of 1 ml of a pure preparation of a single gonococcal antigen in incomplete Freund's adjuvant; the doses were spaced at fortnightly intervals and then the rabbits were bled

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The Surface Properties of Neisseria gonorrhoeae: Isolation of the Major Components of the Outer Membrane

Cited by 106 publications

References 24 publications

Gonococcal outer-membrane protein PIB: comparative sequence analysis and localization of epitopes which are recognized by type-specific and cross-reacting monoclonal antibodies

Gonococcal outer-membrane protein PIB: comparative sequence analysis and localization of epitopes which are recognized by type-specific and cross-reacting monoclonal antibodies

The Isolation of a New Outer Membrane Protein from the Parent Strain of Neisseria gonorrhoeae P 9

Protection by Monospecific Gonococcal Antisera of the Chicken Embryo Challenged With Neisseria Gonorrhoeae

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