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. 1985 Jul;76(1):52–59. doi: 10.1172/JCI111976

Immunogens consisting of oligosaccharides from the capsule of Haemophilus influenzae type b coupled to diphtheria toxoid or the toxin protein CRM197.

P Anderson, M E Pichichero, R A Insel
PMCID: PMC423703  PMID: 3874882

Abstract

Haemophilus influenzae type b (Hib) capsular polysaccharide (PRP) was selectively hydrolyzed to reducing oligosaccharides, and the fraction containing 3-10 ribosylribitolphosphate repeating units (VS) was conjugated by reductive amination to diphtheria toxin (DTx), its nontoxic derivative CRM197 (Dcr), or diphtheria toxoid (DTd). Conjugate DTx-VS retained approximately 1% of native toxicity, which was eliminated by treatment with formalin. Immunization of rabbits with the conjugates elicited antibody (Ab) to PRP and to DTx but not to a model for the linkage determinant. Human adults given single subcutaneous injections had rises in serum Ab to PRP and in bactericidal activity in vitro; the Ab protected infant rats challenged with Hib. Adults had rises also in Ab to DTd, and these Ab protected rabbits against DTx. A series of two injections of the conjugates Dcr-VS and DTd-VS was tested in infants beginning at 19-23 mo of age. Rises in anti-PRP Ab after the primary resembled the rises after PRP vaccine. In contrast to PRP, the conjugates elicited large rises after the secondary vaccinations and a substantial IgG component. Development of bactericidal activity paralleled the rises in anti-PRP Ab. Secondary rises after Dcr-VS were higher than after DTd-VS. In infants 12-16 mo of age, Dcr-VS (but not DTd-VS) elicited strong primary and secondary Ab responses that included IgG and bactericidal activity. Both conjugates produced consistent rises in Ab to DTd.

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Selected References

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  1. Anderson P. Antibody responses to Haemophilus influenzae type b and diphtheria toxin induced by conjugates of oligosaccharides of the type b capsule with the nontoxic protein CRM197. Infect Immun. 1983 Jan;39(1):233–238. doi: 10.1128/iai.39.1.233-238.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson P., Flesher A., Shaw S., Harding A. L., Smith D. H. Phenotypic and genetic variation in the susceptibility of Haemophilus influenzae type b to antibodies to somatic antigens. J Clin Invest. 1980 Apr;65(4):885–891. doi: 10.1172/JCI109741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Anderson P. Intrinsic tritium labeling of the capsular polysaccharide antigen of Haemophilus influenzae type B. J Immunol. 1978 Mar;120(3):866–870. [PubMed] [Google Scholar]
  4. Anderson P., Smith D. H., Ingram D. L., Wilkins J., Wehrle P. F., Howie V. M. Antibody of polyribophate of Haemophilus influenzae type b in infants and children: effect of immunization with polyribophosphate. J Infect Dis. 1977 Aug;136 (Suppl):S57–S62. doi: 10.1093/infdis/136.supplement.s57. [DOI] [PubMed] [Google Scholar]
  5. Anderson P., Smith D. H. Isolation of the capsular polysaccharide from culture supernatant of Haemophilus influenzae type b. Infect Immun. 1977 Feb;15(2):472–477. doi: 10.1128/iai.15.2.472-477.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Anderson P. The protective level of serum antibodies to the capsular polysaccharide of Haemophilus influenzae type b. J Infect Dis. 1984 Jun;149(6):1034–1035. doi: 10.1093/infdis/149.6.1034. [DOI] [PubMed] [Google Scholar]
  7. Aplin J. D., Wriston J. C., Jr Preparation, properties, and applications of carbohydrate conjugates of proteins and lipids. CRC Crit Rev Biochem. 1981;10(4):259–306. doi: 10.3109/10409238109113601. [DOI] [PubMed] [Google Scholar]
  8. Beuvery E. C., Miedema F., van Delft R., Haverkamp J. Preparation and immunochemical characterization of meningococcal group C polysaccharide-tetanus toxoid conjugates as a new generation of vaccines. Infect Immun. 1983 Apr;40(1):39–45. doi: 10.1128/iai.40.1.39-45.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chu C., Schneerson R., Robbins J. B., Rastogi S. C. Further studies on the immunogenicity of Haemophilus influenzae type b and pneumococcal type 6A polysaccharide-protein conjugates. Infect Immun. 1983 Apr;40(1):245–256. doi: 10.1128/iai.40.1.245-256.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Flesher A. R., Insel R. A. Characterization of lipopolysaccharide of Haemophilus influenzae. J Infect Dis. 1978 Dec;138(6):719–730. doi: 10.1093/infdis/138.6.719. [DOI] [PubMed] [Google Scholar]
  11. Goebel W. F. CHEMO-IMMUNOLOGICAL STUDIES ON CONJUGATED CARBOHYDRATE-PROTEINS : XII. THE IMMUNOLOGICAL PROPERTIES OF AN ARTIFICIAL ANTIGEN CONTAINING CELLOBIURONIC ACID. J Exp Med. 1938 Sep 30;68(4):469–484. doi: 10.1084/jem.68.4.469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Granoff D. M., Boies E. G., Munson R. S., Jr Immunogenicity of Haemophilus influenzae type b polysaccharide--diphtheria toxoid conjugate vaccine in adults. J Pediatr. 1984 Jul;105(1):22–27. doi: 10.1016/s0022-3476(84)80350-9. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Lin K. T., Lee C. J. Immune response of neonates to pneumococcal polysaccharide-protein conjugate. Immunology. 1982 Jun;46(2):333–342. [PMC free article] [PubMed] [Google Scholar]
  17. Middaugh J. P. Side effects of diphtheria-tetanus toxoid in adults. Am J Public Health. 1979 Mar;69(3):246–249. doi: 10.2105/ajph.69.3.246. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mäkelä O., Péterfy F., Outschoorn I. G., Richter A. W., Seppälä I. Immunogenic properties of alpha (1----6) dextran, its protein conjugates, and conjugates of its breakdown products in mice. Scand J Immunol. 1984 Jun;19(6):541–550. doi: 10.1111/j.1365-3083.1984.tb00965.x. [DOI] [PubMed] [Google Scholar]
  19. Naor D., Galili N. Immune response to chemically modified antigens. Prog Allergy. 1977;22:107–146. [PubMed] [Google Scholar]
  20. Pappenheimer A. M., Jr, Uchida T., Harper A. A. An immunological study of the diphtheria toxin molecule. Immunochemistry. 1972 Sep;9(9):891–906. doi: 10.1016/0019-2791(72)90163-2. [DOI] [PubMed] [Google Scholar]
  21. Peltola H., Käyhty H., Sivonen A., Mäkelä H. Haemophilus influenzae type b capsular polysaccharide vaccine in children: a double-blind field study of 100,000 vaccinees 3 months to 5 years of age in Finland. Pediatrics. 1977 Nov;60(5):730–737. [PubMed] [Google Scholar]
  22. Robbins J. B. Vaccines for the prevention of encapsulated bacterial diseases: current status, problems and prospects for the future. Immunochemistry. 1978 Nov;15(10-11):839–854. doi: 10.1016/0161-5890(78)90117-7. [DOI] [PubMed] [Google Scholar]
  23. Rojas-Corona R. R., Skarnes R., Tamakuma S., Fine J. The Limulus coagulation test for endotoxin. A comparison with other assay methods. Proc Soc Exp Biol Med. 1969 Nov;132(2):599–601. doi: 10.3181/00379727-132-34268. [DOI] [PubMed] [Google Scholar]
  24. Schneerson R., Barrera O., Sutton A., Robbins J. B. Preparation, characterization, and immunogenicity of Haemophilus influenzae type b polysaccharide-protein conjugates. J Exp Med. 1980 Aug 1;152(2):361–376. doi: 10.1084/jem.152.2.361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]

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