Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1984 Jan;43(1):407–412. doi: 10.1128/iai.43.1.407-412.1984

Conjugation of meningococcal lipopolysaccharide R-type oligosaccharides to tetanus toxoid as route to a potential vaccine against group B Neisseria meningitidis.

H J Jennings, C Lugowski, F E Ashton
PMCID: PMC263442  PMID: 6418661

Abstract

Oligosaccharides were obtained by the mild acid hydrolysis of the lipopolysaccharides from a number of different strains of Neisseria meningitidis, serotypes L2, L3, L4, L5, and L10. The dephosphorylated oligosaccharides were conjugated to tetanus toxoid as their 2-(4-isothiocyanatophenyl)-ethylamine derivatives, which resulted in the incorporation of from 18 to 38 oligosaccharides per molecule of tetanus toxoid. When injected in rabbits, the conjugates produced oligosaccharide-specific antibodies which were predominantly serologically specific but which also exhibited some cross-reactivity. These serological results can be attributed to regions of structural dissimilarity and similarity within the oligosaccharides. The oligosaccharide-specific antibodies were also lipopolysaccharide serotype specific, thus indicating that the oligosaccharides are the determinants associated with this serotype specificity. Consistent with the serological results, the conjugate antisera were bactericidal for the homologous serotype meningococcal organisms and in some cases for heterologous serotype organisms.

Full text

PDF
410

Images in this article

Selected References

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

  1. Cuatrecasas P. Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads. J Biol Chem. 1970 Jun;245(12):3059–3065. [PubMed] [Google Scholar]
  2. Goldschneider I., Gotschlich E. C., Artenstein M. S. Human immunity to the meningococcus. II. Development of natural immunity. J Exp Med. 1969 Jun 1;129(6):1327–1348. doi: 10.1084/jem.129.6.1327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Inman J. K., Dintzis H. M. The derivatization of cross-linked polyacrylamide beads. Controlled introduction of functional groups for the preparation of special-purpose, biochemical adsorbents. Biochemistry. 1969 Oct;8(10):4074–4082. doi: 10.1021/bi00838a026. [DOI] [PubMed] [Google Scholar]
  4. Ito J. I., Jr, Wunderlich A. C., Lyons J., Davis C. E., Guiney D. G., Braude A. I. Role of magnesium in the enzyme-linked immunosorbent assay for lipopolysaccharides of rough Escherichia coli strain J5 and Neisseria gonorrhoeae. J Infect Dis. 1980 Oct;142(4):532–537. doi: 10.1093/infdis/142.4.532. [DOI] [PubMed] [Google Scholar]
  5. Janda J., Work E. A colorimetric estimation of lipopolysaccharides. FEBS Lett. 1971 Sep 1;16(4):343–345. doi: 10.1016/0014-5793(71)80386-1. [DOI] [PubMed] [Google Scholar]
  6. Jennings H. J., Bhattacharjee A. K., Kenne L., Kenny C. P., Calver G. The R-type lipopolysaccharides of Neisseria meningitidis. Can J Biochem. 1980 Feb;58(2):128–136. doi: 10.1139/o80-018. [DOI] [PubMed] [Google Scholar]
  7. Jennings H. J., Hawes G. B., Adams G. A., Kenny C. P. The chemical composition and serological reactions of lipopolysaccharides from serogroups A,B,X, and Y Neisseria meningitidis. Can J Biochem. 1973 Oct;51(10):1347–1354. doi: 10.1139/o73-178. [DOI] [PubMed] [Google Scholar]
  8. Jennings H. J., Johnson K. G., Kenne L. The structure of an R-type oligosaccharide core obtained from some lipopolysaccharides of Neisseria meningitidis. Carbohydr Res. 1983 Sep 16;121:233–241. doi: 10.1016/0008-6215(83)84020-8. [DOI] [PubMed] [Google Scholar]
  9. Jennings H. J., Lugowski C. Immunochemistry of groups A, B, and C meningococcal polysaccharide-tetanus toxoid conjugates. J Immunol. 1981 Sep;127(3):1011–1018. [PubMed] [Google Scholar]
  10. KELLOGG D. S., Jr, PEACOCK W. L., Jr, DEACON W. E., BROWN L., PIRKLE D. I. NEISSERIA GONORRHOEAE. I. VIRULENCE GENETICALLY LINKED TO CLONAL VARIATION. J Bacteriol. 1963 Jun;85:1274–1279. doi: 10.1128/jb.85.6.1274-1279.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Lambden P. R., Heckels J. E. Synthesis of immunogenic oligosaccharide-protein conjugates from the lipopolysaccharide of Neisseria gonorrhoeae P9. J Immunol Methods. 1982;48(2):233–240. doi: 10.1016/0022-1759(82)90197-1. [DOI] [PubMed] [Google Scholar]
  13. Mandrell R. E., Zollinger W. D. Lipopolysaccharide serotyping of Neisseria meningitidis by hemagglutination inhibition. Infect Immun. 1977 May;16(2):471–475. doi: 10.1128/iai.16.2.471-475.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. OUCHTERLONY O. Diffusion-in-gel methods for immunological analysis. II. Prog Allergy. 1962;6:30–154. doi: 10.1159/000313795. [DOI] [PubMed] [Google Scholar]
  15. Schwartz B. A., Gray G. R. Proteins containing reductively aminated disaccharides. Synthesis and chemical characterization. Arch Biochem Biophys. 1977 Jun;181(2):542–549. doi: 10.1016/0003-9861(77)90261-2. [DOI] [PubMed] [Google Scholar]
  16. Svenson S. B., Lindberg A. A. Coupling of acid labile Salmonella specific oligosaccharides to macromolecular carriers. J Immunol Methods. 1979;25(4):323–335. doi: 10.1016/0022-1759(79)90025-5. [DOI] [PubMed] [Google Scholar]
  17. Wyle F. A., Artenstein M. S., Brandt B. L., Tramont E. C., Kasper D. L., Altieri P. L., Berman S. L., Lowenthal J. P. Immunologic response of man to group B meningococcal polysaccharide vaccines. J Infect Dis. 1972 Nov;126(5):514–521. doi: 10.1093/infdis/126.5.514. [DOI] [PubMed] [Google Scholar]
  18. Zollinger W. D., Mandrell R. E., Griffiss J. M., Altieri P., Berman S. Complex of meningococcal group B polysaccharide and type 2 outer membrane protein immunogenic in man. J Clin Invest. 1979 May;63(5):836–848. doi: 10.1172/JCI109383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Zollinger W. D., Mandrell R. E. Outer-membrane protein and lipopolysaccharide serotyping of Neisseria meningitidis by inhibition of a solid-phase radioimmunoassay. Infect Immun. 1977 Nov;18(2):424–433. doi: 10.1128/iai.18.2.424-433.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Zollinger W. D., Mandrell R. E. Type-specific antigens of group A Neisseria meningitidis: lipopolysaccharide and heat-modifiable outer membrane proteins. Infect Immun. 1980 May;28(2):451–458. doi: 10.1128/iai.28.2.451-458.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES