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. 1997 Apr;65(4):1468–1474. doi: 10.1128/iai.65.4.1468-1474.1997

Nonencapsulated Haemophilus influenzae in Aboriginal infants with otitis media: prolonged carriage of P2 porin variants and evidence for horizontal P2 gene transfer.

H C Smith-Vaughan 1, K S Sriprakash 1, J D Mathews 1, D J Kemp 1
PMCID: PMC175155  PMID: 9119489

Abstract

Aboriginal infants in the Northern Territory of Australia experience recurrent otitis media from an early age. Nonencapsulated Haemophilus influenzae (NCHi) colonization of the nasopharynx initially occurs within weeks of birth, persists throughout infancy and most of childhood, and contributes to otitis media. We established previously that the high carriage rates of NCHi in these infants result from concurrent and successive colonization with multiple strains, with sequential elimination of dominant strains. We have now sequenced loops 4, 5, and 6 of the NCHi P2 porin gene and characterized several strains with prolonged carriage times. Furthermore, despite a wide diversity of P2 gene sequences, we have four examples of P2 gene identity for strains with different genetic backgrounds as characterized by PCR ribotyping and randomly amplified polymorphic DNA typing, which leads us to suggest that the P2 gene has been transferred between strains. We also discuss the possibility that the paradoxical observation of cocolonization and prolonged carriage of P2-identical strains is related to immune suppression or tolerance in the host.

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Selected References

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  1. 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]
  2. Bell J., Grass S., Jeanteur D., Munson R. S., Jr Diversity of the P2 protein among nontypeable Haemophilus influenzae isolates. Infect Immun. 1994 Jun;62(6):2639–2643. doi: 10.1128/iai.62.6.2639-2643.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chen Y., Inobe J., Kuchroo V. K., Baron J. L., Janeway C. A., Jr, Weiner H. L. Oral tolerance in myelin basic protein T-cell receptor transgenic mice: suppression of autoimmune encephalomyelitis and dose-dependent induction of regulatory cells. Proc Natl Acad Sci U S A. 1996 Jan 9;93(1):388–391. doi: 10.1073/pnas.93.1.388. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Davies R., Pace R., Jazwinska E. C., Serjeantson S. W. HLA-DR RFLP distributions in two groups of aboriginal Australians. Aust N Z J Med. 1990 Dec;20(6):790–793. doi: 10.1111/j.1445-5994.1990.tb00424.x. [DOI] [PubMed] [Google Scholar]
  5. Duim B., Dankert J., Jansen H. M., van Alphen L. Genetic analysis of the diversity in outer membrane protein P2 of non-encapsulated Haemophilus influenzae. Microb Pathog. 1993 Jun;14(6):451–462. doi: 10.1006/mpat.1993.1044. [DOI] [PubMed] [Google Scholar]
  6. Duim B., van Alphen L., Eijk P., Jansen H. M., Dankert J. Antigenic drift of non-encapsulated Haemophilus influenzae major outer membrane protein P2 in patients with chronic bronchitis is caused by point mutations. Mol Microbiol. 1994 Mar;11(6):1181–1189. doi: 10.1111/j.1365-2958.1994.tb00394.x. [DOI] [PubMed] [Google Scholar]
  7. Falla T. J., Crook D. W., Brophy L. N., Maskell D., Kroll J. S., Moxon E. R. PCR for capsular typing of Haemophilus influenzae. J Clin Microbiol. 1994 Oct;32(10):2382–2386. doi: 10.1128/jcm.32.10.2382-2386.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gardiner D., Hartas J., Currie B., Mathews J. D., Kemp D. J., Sriprakash K. S. Vir typing: a long-PCR typing method for group A streptococci. PCR Methods Appl. 1995 Apr;4(5):288–293. doi: 10.1101/gr.4.5.288. [DOI] [PubMed] [Google Scholar]
  9. Haase E. M., Campagnari A. A., Sarwar J., Shero M., Wirth M., Cumming C. U., Murphy T. F. Strain-specific and immunodominant surface epitopes of the P2 porin protein of nontypeable Haemophilus influenzae. Infect Immun. 1991 Apr;59(4):1278–1284. doi: 10.1128/iai.59.4.1278-1284.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Haase E. M., Yi K., Morse G. D., Murphy T. F. Mapping of bactericidal epitopes on the P2 porin protein of nontypeable Haemophilus influenzae. Infect Immun. 1994 Sep;62(9):3712–3722. doi: 10.1128/iai.62.9.3712-3722.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kehoe M. A., Kapur V., Whatmore A. M., Musser J. M. Horizontal gene transfer among group A streptococci: implications for pathogenesis and epidemiology. Trends Microbiol. 1996 Nov;4(11):436–443. doi: 10.1016/0966-842x(96)10058-5. [DOI] [PubMed] [Google Scholar]
  12. Kroll J. S., Moxon E. R., Loynds B. M. Natural genetic transfer of a putative virulence-enhancing mutation to Haemophilus influenzae type a. J Infect Dis. 1994 Mar;169(3):676–679. doi: 10.1093/infdis/169.3.676. [DOI] [PubMed] [Google Scholar]
  13. Leach A. J., Boswell J. B., Asche V., Nienhuys T. G., Mathews J. D. Bacterial colonization of the nasopharynx predicts very early onset and persistence of otitis media in Australian aboriginal infants. Pediatr Infect Dis J. 1994 Nov;13(11):983–989. doi: 10.1097/00006454-199411000-00009. [DOI] [PubMed] [Google Scholar]
  14. Loos B. G., Bernstein J. M., Dryja D. M., Murphy T. F., Dickinson D. P. Determination of the epidemiology and transmission of nontypable Haemophilus influenzae in children with otitis media by comparison of total genomic DNA restriction fingerprints. Infect Immun. 1989 Sep;57(9):2751–2757. doi: 10.1128/iai.57.9.2751-2757.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Maiden M. C., Malorny B., Achtman M. A global gene pool in the neisseriae. Mol Microbiol. 1996 Sep;21(6):1297–1298. doi: 10.1046/j.1365-2958.1996.981457.x. [DOI] [PubMed] [Google Scholar]
  16. Murphy T. F., Bartos L. C. Human bactericidal antibody response to outer membrane protein P2 of nontypeable Haemophilus influenzae. Infect Immun. 1988 Oct;56(10):2673–2679. doi: 10.1128/iai.56.10.2673-2679.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Relf W. A., Martin D. R., Sriprakash K. S. Antigenic diversity within a family of M proteins from group A streptococci: evidence for the role of frameshift and compensatory mutations. Gene. 1994 Jun 24;144(1):25–30. doi: 10.1016/0378-1119(94)90198-8. [DOI] [PubMed] [Google Scholar]
  18. Sikkema D. J., Murphy T. F. Molecular analysis of the P2 porin protein of nontypeable Haemophilus influenzae. Infect Immun. 1992 Dec;60(12):5204–5211. doi: 10.1128/iai.60.12.5204-5211.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Smith-Vaughan H. C., Leach A. J., Shelby-James T. M., Kemp K., Kemp D. J., Mathews J. D. Carriage of multiple ribotypes of non-encapsulated Haemophilus influenzae in aboriginal infants with otitis media. Epidemiol Infect. 1996 Apr;116(2):177–183. doi: 10.1017/s0950268800052419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Smith-Vaughan H. C., Sriprakash K. S., Mathews J. D., Kemp D. J. Long PCR-ribotyping of nontypeable Haemophilus influenzae. J Clin Microbiol. 1995 May;33(5):1192–1195. doi: 10.1128/jcm.33.5.1192-1195.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Smith H. O., Tomb J. F., Dougherty B. A., Fleischmann R. D., Venter J. C. Frequency and distribution of DNA uptake signal sequences in the Haemophilus influenzae Rd genome. Science. 1995 Jul 28;269(5223):538–540. doi: 10.1126/science.7542802. [DOI] [PubMed] [Google Scholar]
  22. Srikumar R., Dahan D., Gras M. F., Ratcliffe M. J., van Alphen L., Coulton J. W. Antigenic sites on porin of Haemophilus influenzae type b: mapping with synthetic peptides and evaluation of structure predictions. J Bacteriol. 1992 Jun;174(12):4007–4016. doi: 10.1128/jb.174.12.4007-4016.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Vogel L., Duim B., Geluk F., Eijk P., Jansen H., Dankert J., vanAlphen L. Immune selection for antigenic drift of major outer membrane protein P2 of Haemophilus influenzae during persistence in subcutaneous tissue cages in rabbits. Infect Immun. 1996 Mar;64(3):980–986. doi: 10.1128/iai.64.3.980-986.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Williams J. G., Kubelik A. R., Livak K. J., Rafalski J. A., Tingey S. V. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 1990 Nov 25;18(22):6531–6535. doi: 10.1093/nar/18.22.6531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. van Alphen L., Eijk P., Geelen-van den Broek L., Dankert J. Immunochemical characterization of variable epitopes of outer membrane protein P2 of nontypeable Haemophilus influenzae. Infect Immun. 1991 Jan;59(1):247–252. doi: 10.1128/iai.59.1.247-252.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. van Belkum A., Bax R., Peerbooms P., Goessens W. H., van Leeuwen N., Quint W. G. Comparison of phage typing and DNA fingerprinting by polymerase chain reaction for discrimination of methicillin-resistant Staphylococcus aureus strains. J Clin Microbiol. 1993 Apr;31(4):798–803. doi: 10.1128/jcm.31.4.798-803.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. van Belkum A., Duim B., Regelink A., Möller L., Quint W., van Alphen L. Genomic DNA fingerprinting of clinical Haemophilus influenzae isolates by polymerase chain reaction amplification: comparison with major outer-membrane protein and restriction fragment length polymorphism analysis. J Med Microbiol. 1994 Jul;41(1):63–68. doi: 10.1099/00222615-41-1-63. [DOI] [PubMed] [Google Scholar]

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