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. 1986 Oct;78(4):1020–1027. doi: 10.1172/JCI112656

Identification of a 16,600-dalton outer membrane protein on nontypeable Haemophilus influenzae as a target for human serum bactericidal antibody.

T F Murphy, L C Bartos, P A Rice, M B Nelson, K C Dudas, M A Apicella
PMCID: PMC423749  PMID: 2428838

Abstract

A 16,600-D outer membrane protein is present in all strains of Haemophilus influenzae and antibodies to this protein are present in human serum. This study was designed to assess the role of this outer membrane protein (P6) in nontypeable H. influenzae as a target for human serum bactericidal antibody. P6 was isolated and coupled to an affinity column. Depleting normal human serum of antibodies to P6 by affinity chromatography resulted in reduced bactericidal activity of that serum for nontypeable H. influenzae. Immunopurified antibodies to P6 from human serum were bactericidal. Finally, preincubation of bacteria with a monoclonal antibody that recognizes a surface epitope on P6, inhibited human serum bactericidal killing. Taken together, these experiments establish that P6 is a target for human bactericidal antibodies. This observation provides evidence that P6 plays a potentially important role in human immunity to infection by nontypeable H. influenzae.

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Selected References

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  1. Anderson P., Johnston R. B., Jr, Smith D. H. Human serum activities against Hemophilus influenzae, type b. J Clin Invest. 1972 Jan;51(1):31–38. doi: 10.1172/JCI106793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Apicella M. A., Dudas K. C., Campagnari A., Rice P., Mylotte J. M., Murphy T. F. Antigenic heterogeneity of lipid A of Haemophilus influenzae. Infect Immun. 1985 Oct;50(1):9–14. doi: 10.1128/iai.50.1.9-14.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Apicella M. A., Westerink M. A., Morse S. A., Schneider H., Rice P. A., Griffiss J. M. Bactericidal antibody response of normal human serum to the lipooligosaccharide of Neisseria gonorrhoeae. J Infect Dis. 1986 Mar;153(3):520–526. doi: 10.1093/infdis/153.3.520. [DOI] [PubMed] [Google Scholar]
  4. Barenkamp S. J., Munson R. S., Jr, Granoff D. M. Outer membrane protein and biotype analysis of pathogenic nontypable Haemophilus influenzae. Infect Immun. 1982 May;36(2):535–540. doi: 10.1128/iai.36.2.535-540.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Berk S. L., Holtsclaw S. A., Wiener S. L., Smith J. K. Nontypeable Haemophilus influenzae in the elderly. Arch Intern Med. 1982 Mar;142(3):537–539. [PubMed] [Google Scholar]
  6. Dahlberg-Lagergård T. Bactericidal antibodies against noncapsular components of Haemophilus influenzae. J Clin Microbiol. 1983 Mar;17(3):428–431. doi: 10.1128/jcm.17.3.428-431.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Evans F. O., Jr, Sydnor J. B., Moore W. E., Moore G. R., Manwaring J. L., Brill A. H., Jackson R. T., Hanna S., Skaar J. S., Holdeman L. V. Sinusitis of the maxillary antrum. N Engl J Med. 1975 Oct 9;293(15):735–739. doi: 10.1056/NEJM197510092931502. [DOI] [PubMed] [Google Scholar]
  8. Gigliotti F., Insel R. A. Protection from infection with Haemophilus influenzae type b by monoclonal antibody to the capsule. J Infect Dis. 1982 Aug;146(2):249–254. doi: 10.1093/infdis/146.2.249. [DOI] [PubMed] [Google Scholar]
  9. Gnehm H. E., Pelton S. I., Gulati S., Rice P. A. Characterization of antigens from nontypable Haemophilus influenzae recognized by human bactericidal antibodies. Role of Haemophilus outer membrane proteins. J Clin Invest. 1985 May;75(5):1645–1658. doi: 10.1172/JCI111872. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Goldschneider I., Gotschlich E. C., Artenstein M. S. Human immunity to the meningococcus. I. The role of humoral antibodies. J Exp Med. 1969 Jun 1;129(6):1307–1326. doi: 10.1084/jem.129.6.1307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Karasic R. B., Trumpp C. E., Gnehm H. E., Rice P. A., Pelton S. I. Modification of otitis media in chinchillas rechallenged with nontypable Haemophilus influenzae and serological response to outer membrane antigens. J Infect Dis. 1985 Feb;151(2):273–279. doi: 10.1093/infdis/151.2.273. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. Munson R. S., Jr, Granoff D. M. Purification and partial characterization of outer membrane proteins P5 and P6 from Haemophilus influenzae type b. Infect Immun. 1985 Sep;49(3):544–549. doi: 10.1128/iai.49.3.544-549.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Munson R. S., Jr, Shenep J. L., Barenkamp S. J., Granoff D. M. Purification and comparison of outer membrane protein P2 from Haemophilus influenzae type b isolates. J Clin Invest. 1983 Aug;72(2):677–684. doi: 10.1172/JCI111017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Murphy T. F., Apicella M. A. Antigenic heterogeneity of outer membrane proteins of nontypable Haemophilus influenzae is a basis for a serotyping system. Infect Immun. 1985 Oct;50(1):15–21. doi: 10.1128/iai.50.1.15-21.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Murphy T. F., Dudas K. C., Mylotte J. M., Apicella M. A. A subtyping system for nontypable Haemophilus influenzae based on outer-membrane proteins. J Infect Dis. 1983 May;147(5):838–846. doi: 10.1093/infdis/147.5.838. [DOI] [PubMed] [Google Scholar]
  19. Murphy T. F., Nelson M. B., Dudas K. C., Mylotte J. M., Apicella M. A. Identification of a specific epitope of Haemophilus influenzae on a 16,600-dalton outer membrane protein. J Infect Dis. 1985 Dec;152(6):1300–1307. doi: 10.1093/infdis/152.6.1300. [DOI] [PubMed] [Google Scholar]
  20. Musher D. M., Kubitschek K. R., Crennan J., Baughn R. E. Pneumonia and acute febrile tracheobronchitis due to haemophilus influenzae. Ann Intern Med. 1983 Oct;99(4):444–450. doi: 10.7326/0003-4819-99-4-444. [DOI] [PubMed] [Google Scholar]
  21. Myerowitz R. L., Norden C. W., Demchak T. A. Significance of noncapsular antigens in protection against experimental Haemophilus influenzae type b disease: cross-reactivity. Infect Immun. 1978 Aug;21(2):619–626. doi: 10.1128/iai.21.2.619-626.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Norden C. W., Feldman H. A. Hemophilus influenzae, type b, antibody frequencies determined with bactericidal and radioimmunoassay tests. J Clin Microbiol. 1976 Aug;2(2):136–138. [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Peltola H., Käyhty H., Virtanen M., Mäkelä P. H. Prevention of Hemophilus influenzae type b bacteremic infections with the capsular polysaccharide vaccine. N Engl J Med. 1984 Jun 14;310(24):1561–1566. doi: 10.1056/NEJM198406143102404. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Shurin P. A., Pelton S. I., Tager I. B., Kasper D. L. Bactericidal antibody and susceptibility to otitis media caused by nontypable strains of Haemophilus influenzae. J Pediatr. 1980 Sep;97(3):364–369. doi: 10.1016/s0022-3476(80)80182-x. [DOI] [PubMed] [Google Scholar]
  27. Spagnuolo P. J., Ellner J. J., Lerner P. I., McHenry M. C., Flatauer F., Rosenberg P., Rosenthal M. S. Haemophilus influenzae meningitis: the spectrum of disease in adults. Medicine (Baltimore) 1982 Mar;61(2):74–85. [PubMed] [Google Scholar]
  28. Tsang V. C., Wilson B. C., Peralta J. M. Quantitative, single-tube, kinetic-dependent enzyme-linked immunosorbent assay (k-ELISA). Methods Enzymol. 1983;92:391–403. doi: 10.1016/0076-6879(83)92033-5. [DOI] [PubMed] [Google Scholar]
  29. Wald E. R., Milmoe G. J., Bowen A., Ledesma-Medina J., Salamon N., Bluestone C. D. Acute maxillary sinusitis in children. N Engl J Med. 1981 Mar 26;304(13):749–754. doi: 10.1056/NEJM198103263041302. [DOI] [PubMed] [Google Scholar]
  30. Wallace R. J., Jr, Baker C. J., Quinones F. J., Hollis D. G., Weaver R. E., Wiss K. Nontypable Haemophilus influenzae (biotype 4) as a neonatal, maternal, and genital pathogen. Rev Infect Dis. 1983 Jan-Feb;5(1):123–136. doi: 10.1093/clinids/5.1.123. [DOI] [PubMed] [Google Scholar]
  31. Wallace R. J., Jr, Musher D. M., Septimus E. J., McGowan J. E., Jr, Quinones F. J., Wiss K., Vance P. H., Trier P. A. Haemophilus influenzae infections in adults: characterization of strains by serotypes, biotypes, and beta-lactamase production. J Infect Dis. 1981 Aug;144(2):101–106. doi: 10.1093/infdis/144.2.101. [DOI] [PubMed] [Google Scholar]

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