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Journal Article

Fine mapping of outer membrane protein P2 antigenic sites which vary during persistent infection by Haemophilus influenzae.

B Duim, L Vogel, W Puijk, H M Jansen, R H Meloen, J Dankert, L van Alphen
B Duim
Department of Medical Microbiology, University of Amsterdam, The Netherlands.
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L Vogel
Department of Medical Microbiology, University of Amsterdam, The Netherlands.
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W Puijk
Department of Medical Microbiology, University of Amsterdam, The Netherlands.
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H M Jansen
Department of Medical Microbiology, University of Amsterdam, The Netherlands.
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R H Meloen
Department of Medical Microbiology, University of Amsterdam, The Netherlands.
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J Dankert
Department of Medical Microbiology, University of Amsterdam, The Netherlands.
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L van Alphen
Department of Medical Microbiology, University of Amsterdam, The Netherlands.
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ABSTRACT

Antigenic drift of the major outer membrane protein (MOMP) P2 of nonencapsulated Haemophilus influenzae as observed during persistent infections in patients with chronic bronchitis was mimicked in a rabbit model in which H. influenzae persisted in subcutaneous cages. The antigenic drift resulted from amino acid substitutions in potentially surface-exposed loops of MOMP P2. Since in a rabbit model the appearance of antigenic variants was associated with the presence of strain-specific bactericidal antibodies (L. Vogel, B. Duim, F. Geluk, P. Eijk, H. Jansen, J. Dankert, and L. van Alphen, Infect. Immun. 64:980-986, 1996), we determined the epitope specificities of these bactericidal antibodies. The eight loops of MOMP P2 of H. influenzae d1 were separately expressed as fusion proteins with glutathione S-transferase. Sera of rabbits persistently infected with H. influenzae reacted with the loop 5 and loop 6 fusion proteins in immunoblotting and enzyme-linked immunosorbent assay. For fine mapping of the epitopes with pepscan analysis, overlapping synthetic peptides consisting of 12 amino acids were made. Rabbit sera contained antibodies reacting with peptides derived from loop 5 and peptides containing amino acids of the side of loop 6. In addition, MOMP P2 variant-specific reactions with the amino acids located at the tip of loop 6 were detected. The rabbit sera showed variant-specific complement-dependent bactericidal activities, which were eliminated by affinity chromatography with fusion proteins of loop 6 but not of loop 5. We conclude that, during persistence of H. influenzae in rabbits, variant-specific bactericidal antibodies are elicited to the variable tip of MOMP P2 loop 6.

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Fine mapping of outer membrane protein P2 antigenic sites which vary during persistent infection by Haemophilus influenzae.
B Duim, L Vogel, W Puijk, H M Jansen, R H Meloen, J Dankert, L van Alphen
Infection and Immunity Nov 1996, 64 (11) 4673-4679; DOI:

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Fine mapping of outer membrane protein P2 antigenic sites which vary during persistent infection by Haemophilus influenzae.
B Duim, L Vogel, W Puijk, H M Jansen, R H Meloen, J Dankert, L van Alphen
Infection and Immunity Nov 1996, 64 (11) 4673-4679; DOI:
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