Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1990 Jun;58(6):1909–1913. doi: 10.1128/iai.58.6.1909-1913.1990

Expression in Escherichia coli of a high-molecular-weight protective surface antigen found in nontypeable and type b Haemophilus influenzae.

W R Thomas 1, M G Callow 1, R J Dilworth 1, A A Audesho 1
PMCID: PMC258742  PMID: 2187812

Abstract

An Escherichia coli clone producing a high-molecular-weight surface antigen of Haemophilus influenzae type b (Hib) was isolated from a library of Hib DNA fragments cloned as lysogens in a lambda replacement vector. The antigen is found in sarcosyl-insoluble outer membrane protein preparations and was produced by all 36 H. influenzae isolates tested. Absorption studies indicated that the antigen is a surface determinant on all isolates tested. Antibodies to the antigen (D15) were found in eight of nine convalescent-phase sera from children with invasive Hib infection. Affinity-purified antibodies prepared against the cloned antigen gave protection against the development of bacteremia in a rat pup model.

Full text

PDF
1913

Images in this article

Selected References

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

  1. Acute respiratory infections in under-fives: 15 million deaths a year. Lancet. 1985 Sep 28;2(8457):699–701. [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. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  4. Carlone G. M., Thomas M. L., Rumschlag H. S., Sottnek F. O. Rapid microprocedure for isolating detergent-insoluble outer membrane proteins from Haemophilus species. J Clin Microbiol. 1986 Sep;24(3):330–332. doi: 10.1128/jcm.24.3.330-332.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Deich R. A., Metcalf B. J., Finn C. W., Farley J. E., Green B. A. Cloning of genes encoding a 15,000-dalton peptidoglycan-associated outer membrane lipoprotein and an antigenically related 15,000-dalton protein from Haemophilus influenzae. J Bacteriol. 1988 Feb;170(2):489–498. doi: 10.1128/jb.170.2.489-498.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gonzales F. R., Leachman S., Norgard M. V., Radolf J. D., McCracken G. H., Jr, Evans C., Hansen E. J. Cloning and expression in Escherichia coli of the gene encoding the heat-modifiable major outer membrane protein of Haemophilus influenzae type b. Infect Immun. 1987 Dec;55(12):2993–3000. doi: 10.1128/iai.55.12.2993-3000.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hansen E. J., Gonzales F. R., Chamberlain N. R., Norgard M. V., Miller E. E., Cope L. D., Pelzel S. E., Gaddy B., Clausell A. Cloning of the gene encoding the major outer membrane protein of Haemophilus influenzae type b. Infect Immun. 1988 Oct;56(10):2709–2716. doi: 10.1128/iai.56.10.2709-2716.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Holmans P. L., Loftus T. A., Hansen E. J. Cloning and surface expression in Escherichia coli of a structural gene encoding a surface protein of Haemophilus influenzae type b. Infect Immun. 1985 Oct;50(1):236–242. doi: 10.1128/iai.50.1.236-242.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Inzana T. J., Anderson P. Serum factor-dependent resistance of Haemophilus influenzae type b to antibody to lipopolysaccharide. J Infect Dis. 1985 May;151(5):869–877. doi: 10.1093/infdis/151.5.869. [DOI] [PubMed] [Google Scholar]
  10. Kimura A., Gulig P. A., McCracken G. H., Jr, Loftus T. A., Hansen E. J. A minor high-molecular-weight outer membrane protein of Haemophilus influenzae type b is a protective antigen. Infect Immun. 1985 Jan;47(1):253–259. doi: 10.1128/iai.47.1.253-259.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Loeb M. R. Protection of infant rats from Haemophilus influenzae type b infection by antiserum to purified outer membrane protein a. Infect Immun. 1987 Nov;55(11):2612–2618. doi: 10.1128/iai.55.11.2612-2618.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Loeb M. R., Woodin K. A. Cross-reactivity of surface-exposed epitopes of outer membrane antigens of Haemophilus influenzae type b. Infect Immun. 1987 Dec;55(12):2977–2983. doi: 10.1128/iai.55.12.2977-2983.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Munson R., Jr, Grass S. Purification, cloning, and sequence of outer membrane protein P1 of Haemophilus influenzae type b. Infect Immun. 1988 Sep;56(9):2235–2242. doi: 10.1128/iai.56.9.2235-2242.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Nelson M. B., Apicella M. A., Murphy T. F., Vankeulen H., Spotila L. D., Rekosh D. Cloning and sequencing of Haemophilus influenzae outer membrane protein P6. Infect Immun. 1988 Jan;56(1):128–134. doi: 10.1128/iai.56.1.128-134.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Poirier T. P., Kehoe M. A., Beachey E. H. Protective immunity evoked by oral administration of attenuated aroA Salmonella typhimurium expressing cloned streptococcal M protein. J Exp Med. 1988 Jul 1;168(1):25–32. doi: 10.1084/jem.168.1.25. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Shann F., Gratten M., Germer S., Linnemann V., Hazlett D., Payne R. Aetiology of pneumonia in children in Goroka Hospital, Papua New Guinea. Lancet. 1984 Sep 8;2(8402):537–541. doi: 10.1016/s0140-6736(84)90764-5. [DOI] [PubMed] [Google Scholar]
  17. Takala A. K., van Alphen L., Eskola J., Palmgren J., Bol P., Mäkelä P. H. Haemophilus influenzae type b strains of outer membrane subtypes 1 and 1c cause different types of invasive disease. Lancet. 1987 Sep 19;2(8560):647–650. doi: 10.1016/s0140-6736(87)92440-8. [DOI] [PubMed] [Google Scholar]
  18. Thomas W. R., Rossi A. A. Molecular cloning of DNA coding for outer membrane proteins of Haemophilus influenzae type b. Infect Immun. 1986 Jun;52(3):812–817. doi: 10.1128/iai.52.3.812-817.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES