Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1983 Jul;41(1):285–293. doi: 10.1128/iai.41.1.285-293.1983

Induction of active immunity with membrane fractions from Haemophilus influenzae type b.

J P Burans, M Lynn, M Solotorovsky
PMCID: PMC264776  PMID: 6602769

Abstract

Using Escherichia coli strain E-1 as a model, we developed procedures for the preparation of outer- and inner-membrane-enriched fractions as structural units. These procedures could be used to prepare relatively pure inner and outer membrane fractions as determined by succinate dehydrogenase activity, ketodeoxyoctonate levels, and polyacrylamide gradient gel electrophoresis. The use of these procedures to fractionate membrane components from Haemophilus influenzae type b strains H-2 and H-E led to good separation of outer- and inner-membrane-enriched fractions as determined by succinate dehydrogenase and ketodeoxyoctonate levels but incomplete separation as determined by polyacrylamide gradient gel electrophoresis. Although there were differences between the electrophoresis profiles of outer membrane fractions of strains H-2 and H-E, immunization with outer membrane of either strain led to the induction of a high degree of immunoprotection against challenge with the H-2 strain. Protection could also be elicited with inner membrane preparations, but such protection may be due to contamination with outer membrane. Extracted membrane protein induced levels of protection that were comparable to those induced by whole membrane fractions.

Full text

PDF
286

Images in this article

288Image
on p.288
291Image
on p.291
291Image
on p.291

Selected References

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

  1. BUTLER L. O. A defined medium for Haemophilus influenzae and Haemophilus parainfluenzae. J Gen Microbiol. 1962 Jan;27:51–60. doi: 10.1099/00221287-27-1-51. [DOI] [PubMed] [Google Scholar]
  2. Barenkamp S. J., Munson R. S., Jr, Granoff D. M. Subtyping isolates of Haemophilus influenzae type b by outer-membrane protein profiles. J Infect Dis. 1981 May;143(5):668–676. doi: 10.1093/infdis/143.5.668. [DOI] [PubMed] [Google Scholar]
  3. Burans J. P., Kruszewski F. H., Lynn M., Solotorovsky M. Kinetics of Haemophilus influenzae type B infection in normal and ribosome-immunized mice using intraperitoneal and intracerebral routes of inoculation. Br J Exp Pathol. 1981 Oct;62(5):496–503. [PMC free article] [PubMed] [Google Scholar]
  4. Fogel S., Sypherd P. S. Extraction and isolation of individual ribosomal proteins from Escherichia coli. J Bacteriol. 1968 Aug;96(2):358–364. doi: 10.1128/jb.96.2.358-364.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hansen E. J., Frisch C. F., McDade R. L., Jr, Johnston K. H. Identification of immunogenic outer membrane proteins of Haemophilus influenzae type b in the infant rat model system. Infect Immun. 1981 Jun;32(3):1084–1092. doi: 10.1128/iai.32.3.1084-1092.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hansen E. J., Robertson S. M., Gulig P. A., Frisch C. F., Haanes E. J. Immunoprotection of rats against Haemophilus influenzae type B disease mediated by monoclonal antibody against a haemophilus outer-membrane protein. Lancet. 1982 Feb 13;1(8268):366–368. doi: 10.1016/s0140-6736(82)91394-0. [DOI] [PubMed] [Google Scholar]
  7. Katz M., Lynn M., Solotorovsky M. Serological and biological activities of anti-Haemophilus influenzae ribosomal serum. Infect Immun. 1981 Mar;31(3):1125–1131. doi: 10.1128/iai.31.3.1125-1131.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. King S. D., Ramlal A., Wynter H., Moodie K., Castle D., Kuo J. S., Barnes L., Williams C. L. Safety and immunogenicity of a new Haemophilus influenzae type b vaccine in infants under one year of age. Lancet. 1981 Oct 3;2(8249):705–709. doi: 10.1016/s0140-6736(81)91045-x. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. 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]
  11. Loeb M. R., Smith D. H. Outer membrane protein composition in disease isolates of Haemophilus influenzae: pathogenic and epidemiological implications. Infect Immun. 1980 Dec;30(3):709–717. doi: 10.1128/iai.30.3.709-717.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Loeb M. R., Zachary A. L., Smith D. H. Isolation and partial characterization of outer and inner membranes from encapsulated Haemophilus influenzae type b. J Bacteriol. 1981 Jan;145(1):596–604. doi: 10.1128/jb.145.1.596-604.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lynn M., Tewari R. P., Solotorovsky M. Immunoprotective activity of ribosomes from Haemophilus influenzae. Infect Immun. 1977 Feb;15(2):453–460. doi: 10.1128/iai.15.2.453-460.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Osborn M. J., Gander J. E., Parisi E., Carson J. Mechanism of assembly of the outer membrane of Salmonella typhimurium. Isolation and characterization of cytoplasmic and outer membrane. J Biol Chem. 1972 Jun 25;247(12):3962–3972. [PubMed] [Google Scholar]
  15. 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]
  16. Solotorovsky M., Lynn M. Haemophilus influenzae: immunology and immunoprotection. CRC Crit Rev Microbiol. 1978 Sep;6(1):1–32. doi: 10.3109/10408417809090618. [DOI] [PubMed] [Google Scholar]
  17. Tewari R. P., Lynn M., Birnbaum A. J., Solotorovsky M. Characterization of the immunoprotective antigen of ribosomal preparations from Haemophilus influenzae. Infect Immun. 1978 Jan;19(1):58–65. doi: 10.1128/iai.19.1.58-65.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. WEISSBACH A., HURWITZ J. The formation of 2-keto-3-deoxyheptonic acid in extracts of Escherichia coli B. I. Identification. J Biol Chem. 1959 Apr;234(4):705–709. [PubMed] [Google Scholar]
  19. Yamato I., Anraku Y., Hirosawa K. Cytoplasmic membrane vesicles of Escherichia coli. A simple method for preparing the cytoplasmic and outer membranes. J Biochem. 1975 Apr;77(4):705–718. doi: 10.1093/oxfordjournals.jbchem.a130774. [DOI] [PubMed] [Google Scholar]

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

RESOURCES