Intra-strain heterogeneity of gonococcal pili is related to opacity colony variance.

Salit, Irving E.; Blake, Milan; Gotschlich, Emil C.

doi:10.1084/jem.151.3.716

Cited by 58 publications

(29 citation statements)

References 26 publications

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“…Cultures were heated at 60°C for 30 min, after which the cells were pelleted by centrifugation (3,000 g, 30 min, 4°C). The supernatant was examined for pili content by 12.5% SDS-PAGE using a modification of the method of Laemmli (15,16). K99 pili were purified by adjusting the culture supernatant to pH 9.7 with 10 N NaOH and stirring at room temperature (RT) for 10 min .…”

Section: Methodsmentioning

confidence: 99%

Oral vaccination. Identification of classes of proteins that provoke an immune response upon oral feeding.

Aizpurua¹,

Russell-Jones²

1988

The Journal of Experimental Medicine

204

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Oral immunization of an animal is generally hard to achieve unless large quantities of antigen are administered. In this study a number of antigens were tested for their ability to elicit a systemic immune response upon oral administration. It was found that bacterial pili, LTB, lectins, and a viral hemagglutinin were all able to elicit significant antibody titers upon oral feeding. The immune response thus generated to LTB and K99 pili could be completely abolished by cofeeding a number of sugars that have close structural homology to the terminal sugars of the GM1 and GM2 gangliosides to which these molecules are known to bind. All of the proteins that were active in oral immunization are known to possess "lectin or lectin-like" binding activities. It is therefore proposed that these molecules are able to bind to glycolipids and glycoproteins on the intestinal mucosa and to stimulate these cells to transport the proteins into the systemic circulation, thereby eliciting a systemic immune response. Molecules that did not possess this binding activity were unable to elicit significant responses at the doses tested.

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Section: Methodsmentioning

confidence: 99%

Oral vaccination. Identification of classes of proteins that provoke an immune response upon oral feeding.

Aizpurua¹,

Russell-Jones²

1988

The Journal of Experimental Medicine

204

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“…The modifications and sample preparations of the Laemmli SDS-PAGE method (16) using a discontinuous buffer system has been described in detail elsewhere (17).…”

Section: Sodium Dodecyl Sulfate-polyacrylamide Gel Electrophoresis (Smentioning

confidence: 99%

“…One method described by Ames (18) determines the isoelectric point in denaturing conditions and was performed using polyacrylamide as the supporting material. The samples were prepared in boiling 2% SDS and 8 M urea and applied to the acidic end, as described previously (17). The isoelectric point of Protein II in non-1 Abbreviations used in this paper: GLC, gas liquid chromatography; LPS, lipopolysaccharide; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis.…”

Section: Sodium Dodecyl Sulfate-polyacrylamide Gel Electrophoresis (Smentioning

confidence: 99%

Purification and partial characterization of the opacity-associated proteins of Neisseria gonorrhoeae.

Blake¹,

Gotschlich²

1984

The Journal of Experimental Medicine

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Gonococci, grown on agar, frequently give rise to opaque colonies. This opacity phenotype is associated with the presence of one or more outer membrane proteins of approximately 28,000 mol weight. These proteins are included within a class of proteins named proteins II. A method is described to isolate and purify the opacity-associated proteins from Neisseria gonorrhoeae. This method uses high concentrations of calcium and a zwitterionic detergent at pH 4.0. Under these conditions proteins II are readily solubilized from the outer membrane. Further purification is achieved by ion exchange and molecular sieve chromatography in the presence of the zwitterionic detergent. The opacity-associated proteins are very basic with isoelectric points varying between 9.0 to 10.0. Further evidence for their basic nature is their behavior on ion exchange chromatography and their amino acid composition.

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“…Recent work has shown that even in a single strain surface antigens are capable of considerable variations in vitro (Swanson, 1978; and probably in vivo (James & Swanson, 1977;McBride et al, 1981). Colonial variants of the same strain have been shown to produce pili with altered morphology and subunit molecular weight (Penn et al, 1980;Salit et al, 1980), which show differences in their attachment to epithelial cells (Lambden et al, 1981 b). However, the proteins present in the outer membrane appear capable of even greater variations.…”

Section: Introductionmentioning

confidence: 99%

Antigenic Variation of Outer Membrane Protein II in Colonial Variants of Neisseria gonorrhoeae P9

Diaz

Heckels

1982

Microbiology

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Antibodies were detected by an enzyme-linked immunosorbent assay (ELISA) in sera from rabbits immunized with outer membranes from colonial opacity variants of Neisseria gonorrhoeae P9. ELISA-inhibition experiments with purified antigens revealed approximately equal proportions of antibodies directed against each of the three major surface antigens, lipopolysaccharide, the major outer membrane protein (protein I) and protein 11, the variable protein associated with colonial opacity. Inhibition experiments with intact gonococci showed considerable surface antigenic diversity which could be correlated with differences between the protein I1 species present. Despite their considerable structural homology, different protein I1 species from colonial variants of the same strain showed little cross-reactivity with specific anti-protein I1 sera, thus demonstrating the considerable variation in that part of the antigen which is exposed on the surface of the gonococcus and is closely involved in pathogenic mechanisms.

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Intra-strain heterogeneity of gonococcal pili is related to opacity colony variance.

Cited by 58 publications

References 26 publications

Oral vaccination. Identification of classes of proteins that provoke an immune response upon oral feeding.

Oral vaccination. Identification of classes of proteins that provoke an immune response upon oral feeding.

Purification and partial characterization of the opacity-associated proteins of Neisseria gonorrhoeae.

Antigenic Variation of Outer Membrane Protein II in Colonial Variants of Neisseria gonorrhoeae P9

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