Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1981 Jun;32(3):985–994. doi: 10.1128/iai.32.3.985-994.1981

Antigen-specific serotyping of Neisseria gonorrhoeae: characterization based upon principal outer membrane protein.

T M Buchanan, J F Hildebrandt
PMCID: PMC351548  PMID: 6166568

Abstract

Principal outer membrane protein (protein I) of Neisseria gonorrhoeae was prepared nearly free of lipopolysaccharide (LPS) and substantially purified from other membrane proteins by chromatography of partially purified gonococcal outer membranes over Sepharose 6B in the presence of deoxycholate at pH 9.0. This protein I of nine separate antigenic types was coated to polystyrene tubes and used in the enzyme-linked immunosorbent assay (ELISA) to measure antibody to protein I or in inhibition tests to quantitate protein I antigen. No significant inhibition of the ELISA test was produced by purified LPS from the strain used to prepare each of the protein I types or by whole gonococci bearing the same LPS but different protein I antigens as the strain used to produce a given protein I antigen. Of 125 strains of gonococci used as whole organisms to inhibit the protein I ELISA, 124 (99%) typed with one or more of the nine protein I types, and 35% of these typed with a single protein I serotype. Sixty-one of 65 (94%) strains from Seattle and Atlanta patients with disseminated gonococcal infection contained protein I serotype 1, and 16 of 24 (64%) strains from Seattle patients with salpingitis bore one or both of protein I serotypes 1 and 2.

Full text

PDF
985

Images in this article

987Image
on p.987
988Image
on p.988
988Image
on p.988
990Image
on p.990

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. AMES B. N., DUBIN D. T. The role of polyamines in the neutralization of bacteriophage deoxyribonucleic acid. J Biol Chem. 1960 Mar;235:769–775. [PubMed] [Google Scholar]
  2. Apicella M. A. Serogrouping of Neisseria gonorrhoeae: identification of four immunologically distinct acidic polysaccharides. J Infect Dis. 1976 Oct;134(4):377–383. doi: 10.1093/infdis/134.4.377. [DOI] [PubMed] [Google Scholar]
  3. Arko R. J., Kraus S. J., Brown W. J., Buchanan T. M., Kuhn U. S. Neisseria gonorrhoeae: effects of systemic immunization on resistance of chimpanzees to urethral infection. J Infect Dis. 1974 Aug;130(2):160–164. doi: 10.1093/infdis/130.2.160. [DOI] [PubMed] [Google Scholar]
  4. Buchanan T. M. Antigen-specific serotyping of Neisseria gonorrhoeae. I. Use of an enzyme-linked immunosorbent assay to quantitate pilus antigens on gonococci. J Infect Dis. 1978 Sep;138(3):319–315. doi: 10.1093/infdis/138.3.319. [DOI] [PubMed] [Google Scholar]
  5. Buchanan T. M. Antigenic heterogeneity of gonococcal pili. J Exp Med. 1975 Jun 1;141(6):1470–1475. doi: 10.1084/jem.141.6.1470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Buchanan T. M., Arko R. J. Immunity to gonococcal infection induced by vaccination with isolated outer membranes of Neisseria gonorrhoeae in guinea pigs. J Infect Dis. 1977 Jun;135(6):879–887. doi: 10.1093/infdis/135.6.879. [DOI] [PubMed] [Google Scholar]
  7. Buchanan T. M., Eschenbach D. A., Knapp J. S., Holmes K. K. Gonococcal salpingitis is less likely to recur with Neisseria gonorrhoeae of the same principal outer membrane protein antigenic type. Am J Obstet Gynecol. 1980 Dec 1;138(7 Pt 2):978–980. doi: 10.1016/0002-9378(80)91091-1. [DOI] [PubMed] [Google Scholar]
  8. Buchanan T. M., Pearce W. A. Pili as a mediator of the attachment of gonococci to human erythrocytes. Infect Immun. 1976 May;13(5):1483–1489. doi: 10.1128/iai.13.5.1483-1489.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Buchanan T. M., Pearce W. A., Schoolnik G. K., Arko R. J. Protection against infection with Neisseria gonorrhoeae by immunization with outer membrane protein complex and purified pili. J Infect Dis. 1977 Aug;136 (Suppl):S132–S137. doi: 10.1093/infdis/136.supplement.s132. [DOI] [PubMed] [Google Scholar]
  10. Chen N. C., Kamel K., Zuckerman J., Gaafar H. A. Purification of Neisseria gonorrhoeae surface L-antigen. Infect Immun. 1977 Oct;18(1):230–236. doi: 10.1128/iai.18.1.230-236.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. GREENWOOD F. C., HUNTER W. M., GLOVER J. S. THE PREPARATION OF I-131-LABELLED HUMAN GROWTH HORMONE OF HIGH SPECIFIC RADIOACTIVITY. Biochem J. 1963 Oct;89:114–123. doi: 10.1042/bj0890114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Green M. R., Pastewka J. V., Peacock A. C. Differential staining of phosphoproteins on polyacrylamide gels with a cationic carbocyanine dye. Anal Biochem. 1973 Nov;56(1):43–51. doi: 10.1016/0003-2697(73)90167-x. [DOI] [PubMed] [Google Scholar]
  13. Heckels J. E. The surface of Neisseria gonorrhoeae: isolation of the major components of the outer membrane. J Gen Microbiol. 1977 Apr;99(2):333–341. doi: 10.1099/00221287-99-2-333. [DOI] [PubMed] [Google Scholar]
  14. Hildebrandt J. F., Mayer L. W., Wang S. P., Buchanan T. M. Neisseria gonorrhoeae acquire a new principal outer-membrane protein when transformed to resistance to serum bactericidal activity. Infect Immun. 1978 Apr;20(1):267–272. doi: 10.1128/iai.20.1.267-272.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Holmes K. K., Counts G. W., Beaty H. N. Disseminated gonococcal infection. Ann Intern Med. 1971 Jun;74(6):979–993. doi: 10.7326/0003-4819-74-6-979. [DOI] [PubMed] [Google Scholar]
  16. Johnston K. H., Gotschlich E. C. Isolation and characterization of the outer membrane of Neisseria gonorrhoeae. J Bacteriol. 1974 Jul;119(1):250–257. doi: 10.1128/jb.119.1.250-257.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Johnston K. H., Holmes K. K., Gotschlich E. C. The serological classification of Neisseria gonorrhoeae. I. Isolation of the outer membrane complex responsible for serotypic specificity. J Exp Med. 1976 Apr 1;143(4):741–758. doi: 10.1084/jem.143.4.741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. KEYSER J. W. STAINING OF SERUM GLYCOPROTEINS AFTER ELECTROPHORETIC SEPARATION IN ACRYLAMIDE GELS. Anal Biochem. 1964 Oct;9:249–252. doi: 10.1016/0003-2697(64)90111-3. [DOI] [PubMed] [Google Scholar]
  19. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  20. McDade R. L., Jr, Johnston K. H. Characterization of serologically dominant outer membrane proteins of Neisseria gonorrhoeae. J Bacteriol. 1980 Mar;141(3):1183–1191. doi: 10.1128/jb.141.3.1183-1191.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Novotny P., Turner W. H. Immunological heterogeneity of pili of Neisseria gonorrhoeae. J Gen Microbiol. 1975 Jul;89(1):87–92. doi: 10.1099/00221287-89-1-87. [DOI] [PubMed] [Google Scholar]
  22. OSBORN M. J. STUDIES ON THE GRAM-NEGATIVE CELL WALL. I. EVIDENCE FOR THE ROLE OF 2-KETO- 3-DEOXYOCTONATE IN THE LIPOPOLYSACCHARIDE OF SALMONELLA TYPHIMURIUM. Proc Natl Acad Sci U S A. 1963 Sep;50:499–506. doi: 10.1073/pnas.50.3.499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pearce W. A., Buchanan T. M. Attachment role of gonococcal pili. Optimum conditions and quantitation of adherence of isolated pili to human cells in vitro. J Clin Invest. 1978 Apr;61(4):931–943. doi: 10.1172/JCI109018. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rice P. A., Kasper D. L. Characterization of gonococcal antigens responsible for induction of bactericidal antibody in disseminated infection. J Clin Invest. 1977 Nov;60(5):1149–1158. doi: 10.1172/JCI108867. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Schoolnik G. K., Buchanan T. M., Holmes K. K. Gonococci causing disseminated gonococcal infection are resistant to the bactericidal action of normal human sera. J Clin Invest. 1976 Nov;58(5):1163–1173. doi: 10.1172/JCI108569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. WHITE L. A., KELLOGG D. S., Jr NEISSERIA GONORRHOEAE IDENTIFICATION IN DIRECT SMEARS BY A FLUORESCENT ANTIBODY-COUNTERSTAIN METHOD. Appl Microbiol. 1965 Mar;13:171–174. doi: 10.1128/am.13.2.171-174.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wang S. P., Holmes K. K., Knapp J. S., Ott S., Kyzer D. D. Immunologic classification of Neisseria gonorrhoeae with micro-immunofluorescence. J Immunol. 1977 Sep;119(3):795–803. [PubMed] [Google Scholar]
  28. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES