Abstract
Monoclonal antibodies (MAbs) were elicited to the nontypeable Haemophilus influenzae variants d1 to d4, which differ in the outer membrane protein P2 to analyze the immunological properties of the variable parts of this protein. Five MAbs reacted in a whole-cell enzyme-linked immunosorbent assay (ELISA) only with the homologous strain and in some cases with its variants, but not with 69 unrelated nonencapsulated H. influenzae isolates; nine MAbs also reacted with some other H. influenzae isolates, and four MAbs showed broad cross-reactivity. All of the MAbs reacted with purified protein P2 in ELISAs and immunoblotting. The five MAbs which reacted with the homologous strain d3 and not with the variants d1, d2, and d4 promoted complement-dependent bactericidal activity against strain d3. These and four other MAbs reacted with the intact bacteria of strain d3 in immunogold electron microscopy, indicating that they were directed against surface-exposed epitopes of outer membrane protein P2. A mutant of strain d3 was isolated as a survivor from bacterial killing by complement and MAb 30DA5. This mutant had an altered P2 protein on sodium dodecyl sulfate-polyacrylamide gels and had lost its reactivity with all of the five H. influenzae d3-specific MAbs but not with the other MAbs. From these results, we conclude that the variable parts of outer membrane protein P2 of nonencapsulated H. influenzae from the sputum of patients with chronic obstructive pulmonary disease are immunogenic and mostly surface exposed. Only strain-specific MAbs promoted complement-dependent killing of the bacteria, which was abolished in a spontaneous mutant with an altered P2 protein.
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Selected References
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- Abdillahi H., Poolman J. T. Neisseria meningitidis group B serosubtyping using monoclonal antibodies in whole-cell ELISA. Microb Pathog. 1988 Jan;4(1):27–32. doi: 10.1016/0882-4010(88)90045-9. [DOI] [PubMed] [Google Scholar]
- 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]
- Barenkamp S. J., Shurin P. A., Marchant C. D., Karasic R. B., Pelton S. I., Howie V. M., Granoff D. M. Do children with recurrent Haemophilus influenzae otitis media become infected with a new organism or reacquire the original strain? J Pediatr. 1984 Oct;105(4):533–537. doi: 10.1016/s0022-3476(84)80415-1. [DOI] [PubMed] [Google Scholar]
- Groeneveld K., Eijk P. P., van Alphen L., Jansen H. M., Zanen H. C. Haemophilus influenzae infections in patients with chronic obstructive pulmonary disease despite specific antibodies in serum and sputum. Am Rev Respir Dis. 1990 May;141(5 Pt 1):1316–1321. doi: 10.1164/ajrccm/141.5_Pt_1.1316. [DOI] [PubMed] [Google Scholar]
- Groeneveld K., van Alphen L., Eijk P. P., Jansen H. M., Zanen H. C. Changes in outer membrane proteins of nontypable Haemophilus influenzae in patients with chronic obstructive pulmonary disease. J Infect Dis. 1988 Aug;158(2):360–365. doi: 10.1093/infdis/158.2.360. [DOI] [PubMed] [Google Scholar]
- Groeneveld K., van Alphen L., Eijk P. P., Visschers G., Jansen H. M., Zanen H. C. Endogenous and exogenous reinfections by Haemophilus influenzae in patients with chronic obstructive pulmonary disease: the effect of antibiotic treatment on persistence. J Infect Dis. 1990 Mar;161(3):512–517. doi: 10.1093/infdis/161.3.512. [DOI] [PubMed] [Google Scholar]
- Groeneveld K., van Alphen L., Voorter C., Eijk P. P., Jansen H. M., Zanen H. C. Antigenic drift of Haemophilus influenzae in patients with chronic obstructive pulmonary disease. Infect Immun. 1989 Oct;57(10):3038–3044. doi: 10.1128/iai.57.10.3038-3044.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Klebba P. E., Benson S. A., Bala S., Abdullah T., Reid J., Singh S. P., Nikaido H. Determinants of OmpF porin antigenicity and structure. J Biol Chem. 1990 Apr 25;265(12):6800–6810. [PubMed] [Google Scholar]
- 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]
- Lugtenberg B., Van Alphen L. Molecular architecture and functioning of the outer membrane of Escherichia coli and other gram-negative bacteria. Biochim Biophys Acta. 1983 Mar 21;737(1):51–115. doi: 10.1016/0304-4157(83)90014-x. [DOI] [PubMed] [Google Scholar]
- 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]
- Murphy T. F., Apicella M. A. Nontypable Haemophilus influenzae: a review of clinical aspects, surface antigens, and the human immune response to infection. Rev Infect Dis. 1987 Jan-Feb;9(1):1–15. doi: 10.1093/clinids/9.1.1. [DOI] [PubMed] [Google Scholar]
- Murphy T. F., Bernstein J. M., Dryja D. M., Campagnari A. A., Apicella M. A. Outer membrane protein and lipooligosaccharide analysis of paired nasopharyngeal and middle ear isolates in otitis media due to nontypable Haemophilus influenzae: pathogenetic and epidemiological observations. J Infect Dis. 1987 Nov;156(5):723–731. doi: 10.1093/infdis/156.5.723. [DOI] [PubMed] [Google Scholar]
- Vachon V., Lyew D. J., Coulton J. W. Transmembrane permeability channels across the outer membrane of Haemophilus influenzae type b. J Bacteriol. 1985 Jun;162(3):918–924. doi: 10.1128/jb.162.3.918-924.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van Alphen L., Riemens T., Poolman J., Hopman C., Zanen H. C. Homogeneity of cell envelope protein subtypes, lipopolysaccharide serotypes, and biotypes among Haemophilus influenzae type b from patients with meningitis in The Netherlands. J Infect Dis. 1983 Jul;148(1):75–81. doi: 10.1093/infdis/148.1.75. [DOI] [PubMed] [Google Scholar]
- van Alphen L., Riemens T., Poolman J., Zanen H. C. Characteristics of major outer membrane proteins of Haemophilus influenzae. J Bacteriol. 1983 Aug;155(2):878–885. doi: 10.1128/jb.155.2.878-885.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van Alphen L., van den Berghe N., Geelen-van den Broek L. Interaction of Haemophilus influenzae with human erythrocytes and oropharyngeal epithelial cells is mediated by a common fimbrial epitope. Infect Immun. 1988 Jul;56(7):1800–1806. doi: 10.1128/iai.56.7.1800-1806.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van Ham S. M., Mooi F. R., Sindhunata M. G., Maris W. R., van Alphen L. Cloning and expression in Escherichia coli of Haemophilus influenzae fimbrial genes establishes adherence to oropharyngeal epithelial cells. EMBO J. 1989 Nov;8(11):3535–3540. doi: 10.1002/j.1460-2075.1989.tb08519.x. [DOI] [PMC free article] [PubMed] [Google Scholar]