Protection against Infection with Neisseria gonorrhoeae by Immunization with Outer Membrane Protein Complex and Purified Pili

Buchanan, Thomas M.; Pearce, William A.; Schoolnik, Gary K.; Arko, R J

doi:10.1093/infdis/136.supplement.s132

Cited by 42 publications

(30 citation statements)

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“…The resulting protein contained < 4 % LPS by weight compared with total protein, as judged by assay for KDO, and consisted of > 90 % POMP on SDS-polyacrylamide gel electrophoresis (Buchanan et al, 1977 ;. The LPS used for competitive inhibition experiments, for rabbit immunization and for coating ELISA tubes was isolated from gonococcal outer membrane preparations by suspending them in a 1.5 % (w/v) sodium deoxycholate, 5 m-EDTA, 50 m-glycine buffer, pH 9.0, followed by chromatography on a Sepharose 6B (Pharmacia) column as described for the isolation of POMP .…”

Section: Methodsmentioning

confidence: 99%

“…Potential roles have been suggested for bactericidal antibodies (Brooks et al, 1976;Buchanan et al, 1978;Kasper et al, 1977;McCutchan et al, 1976;, for local secretory antibodies blocking attachment (O'Reilly et a!., 1976;Tramont, 1977), and for opsonic antibodies (Ofek et al, 1974;Penn et al, 1977). That the latter might play a significant role is supported by the observation that virulent Type 1 organisms are more resistant to phagocytosis (Blake & Swanson, 1975;Dilworth et al, 1975;Punsalang & Sawyer, 1973) and that opsonic antibody is acquired in some patients with gonorrhoea (Bisno et al, 1975).…”

Section: Introductionmentioning

confidence: 99%

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Immune-enhanced Phagocytosis of Neisseria gonorrhoeae by Macrophages: Characterization of the Major Antigens to which Opsonins are Directed

et al. 1980

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Antisera were prepared in rabbits against whole organisms of colony type 1 Neisseria gonorrhoeae strains F62 and B (from gonococcal urethritis) and 7122 (a strain typical of those associated with disseminated gonococcal infection), and against purified outer membrane components from the same strains including pili and principal outer membrane protein. Antibody levels to pilj, principal outer membrane protein and lipopolysaccharide were determined using a quantitative enzyme-linked immunosorbent assay. Each antiserum was heat-inactivated and tested for opsonic activity in a quantitative assay of phagocytosis. Each whole-organism antiserum was opsonic for its homologous strain, and this immuneenhanced phagocytosis was decreased by adsorption with homologous purified outer membrane components : pili B lipopolysaccharide P principal outer membrane protein.Opsonic activity was approximately equal for antiserum to purified pili and antiserum to the whole organisms for each of the three strains, and purified antibody to pili was highly opsonic. The F(ab'), fragments of antibody to pili were not opsonic, indicating a role for the Fc receptor on the phagocyte membrane in immune-enhanced phagocytosis of gonococci.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Immune-enhanced Phagocytosis of Neisseria gonorrhoeae by Macrophages: Characterization of the Major Antigens to which Opsonins are Directed

et al. 1980

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“…Type IV pili promote initial attachment of GC to the epithelial cell (8,24,25,53; J. G. Cannon, M. S. Cohen, S. F. Isbey, T. L. Snodgrass, A. Wallace, J. A. F. Dempsey, M. Apicella, and D. Zhou, Abstr.…”

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confidence: 99%

Isolation of Neisseria gonorrhoeae Mutants That Show Enhanced Trafficking across Polarized T84 Epithelial Monolayers

et al. 2000

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Initiation of a gonococcal infection involves attachment of Neisseria gonorrhoeae to the plasma membrane of an epithelial cell in the mucosal epithelium and its internalization, transepithelial trafficking, and exocytosis from the basal membrane. Piliation and expression of certain Opa proteins and the immunoglobulin A1 protease influence the transcytosis process. We are interested in identifying other genetic determinants of N. gonorrhoeae that play a role in transcellular trafficking. Using polarized T84 monolayers as a model epithelial barrier, we have assayed an N. gonorrhoeae FA1090 minitransposon (mTn) mutant bank for isolates that traverse the monolayer more quickly than the isogenic wild-type (WT) strain. From an initial screen, we isolated four mutants, defining three genetic loci, that traverse monolayers significantly more quickly than their WT parent strain. These mutants adhere to and invade cells normally and do not affect the integrity of the monolayer barrier. Backcrosses of the mutations into the WT FA1090 strain yielded mutants with a similar fast-trafficking phenotype. In two mutants, the mTns had inserted 370 bp apart into the same locus, which we have named fit, for fast intracellular trafficker. Backcrosses of one of these mutants into the MS11A genetic background also yielded a fast-trafficking mutant. The fit locus contains two overlapping open reading frames, fitA and fitB, whose deduced amino acid sequences have predicted molecular weights of 8.6 and 15.3, respectively. Neither protein contains a signal sequence. FitA has a potential helix-turn-helix motif, while the deduced sequence of FitB offers no clues to its function. fitA or fitB homologues are present in the genomes of Pseudomonas syringae and Rhizobium meliloti, but not Neisseria meningitidis. Replication of the MS11A fitA mutant in A431 and T84 cells is significantly accelerated compared to that of the isogenic WT strain. In contrast, growth of this mutant in liquid media is normal. Taken together, these results strongly suggest that traversal of N. gonorrhoeae across an epithelial barrier is linked to intracellular bacterial growth.

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“…al., 1974;Ward, Watt and Robertson, 1974;Mhrdh and Westrom, 1976). Antibodies to gonococcal pili are only weakly bactericidal (Buchanan et al, 1977), their main mode of action in vitro being to block attachment to substrate cells (Buchanan and Pearce, 1976;Tramont, 1976). Pili have also been associated with the inhibition of phagocytosis (Blake and Swanson, 1975).…”

Section: Discussionmentioning

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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Protection against Infection with Neisseria gonorrhoeae by Immunization with Outer Membrane Protein Complex and Purified Pili

Cited by 42 publications

References 5 publications

Immune-enhanced Phagocytosis of Neisseria gonorrhoeae by Macrophages: Characterization of the Major Antigens to which Opsonins are Directed

Immune-enhanced Phagocytosis of Neisseria gonorrhoeae by Macrophages: Characterization of the Major Antigens to which Opsonins are Directed

Isolation of Neisseria gonorrhoeae Mutants That Show Enhanced Trafficking across Polarized T84 Epithelial Monolayers

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

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