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Quantifying the Two P6 Populations in Various Strains of Nontypable Haemophilus influenzae
Author(s) -
Shaul Jeffrey,
Reulbach Casey,
Kisselstein Breanne,
Pichichero Michael,
Michel Lea
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.571.11
Subject(s) - haemophilus influenzae , bacterial outer membrane , microbiology and biotechnology , population , periplasmic space , biology , immunology , virology , medicine , escherichia coli , genetics , gene , environmental health , antibiotics
Nontypeable Haemophilus influenzae (NTHi) is a pathogenic bacterium that causes acute otitis media (ear infections) and nasopharyngeal infections. There is a commonly used vaccine against the type B form of Hi (Hib), but there is no vaccine to protect against NTHi in the United States. The lipoprotein P6 found on the outer membrane (OM) of NTHi is currently one of the leading protein vaccine candidates for protection against NTHi. However, research from our lab has shown that P6 can be inserted into the OM in two different orientations: attached to the inner leaflet of the OM and facing in toward the periplasm or attached to the outer leaflet of the OM and facing out toward the extracellular space. When P6 is in its external orientation, it can interact with antibodies, making it a suitable target for vaccines. In this study, we used a biotinylation labeling technique and immunoblotting to separate and quantify the external (surface exposed) P6 and internal P6. The results of our study suggest that in the clinical strains of NTHi that we tested, only a minor population of P6 is surface exposed. We consider the possible ramifications of minimal surface exposure on P6′s vaccine candidacy.