Antibodies directed at a conserved motif in loop 6 of outer membrane protein P2 of nontypeable Haemophilus influenzae recognize multiple strains in immunoassays.
The P2 porin is the most abundant protein in the outer membrane of nontypeable Haemophilus influenzae. Analysis of P2 sequences from a limited number of strains reveals the presence of both heterogeneous and conserved surface-exposed loops of the P2 molecule among strains. We have previously shown that antibodies raised against the loop 6 sequence of P2 from strain 5657 are bactericidal against multiple isolates. In this study, we determined the nucleotide sequence of the loop 6 region of the P2 molecule from 108 strains of nontypeable H. influenzae in order to assess more rigorously the degree of conservation of loop 6. Based on this analysis, we identified a conserved sequence, different from that of strain 5657, that occurs in approximately one-third of the strains sequenced. To assess the potential of this peptide as a vaccine antigen, antibodies raised to a multiple antigenic peptide corresponding to this sequence were characterized with respect to specificity for the P2 molecule and reactivity with heterologous strains in immunoblot assay, flow cytometry and bactericidal assays. Antibodies were reactive to the P2 molecule of 16 of 20 strains tested by immunoblot assay. Antibodies recognized nine of the 20 strains in a flow cytometry assay, and 13 of 20 demonstrated complement-mediated killing in bactericidal assays. These results support the concept of using conserved regions of the P2 protein as a vaccine antigen.
Department of Microbiology, University of Buffalo, State University of New York, Buffalo, NY, USA.
John M
JM
Neary
Timothy F
TF
Murphy
D000328:Adult; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000276:immunology*; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D001770:Blood Bactericidal Activity; D002648:Child; D017124:Conserved Sequence; D005434:Flow Cytometry; D006193:Haemophilus influenzae / Q000145:classification* / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007118:Immunoassay; D007223:Infant; D008969:Molecular Sequence Data; D009154:Mutation; D018272:Porins / Q000235:genetics / Q000276:immunology*; D017422:Sequence Analysis, DNA; D013045:Species Specificity
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D018272:Porins; C451502:ompP2 protein, Haemophilus influenzae