Immunologic and structural relationships of the minor pilus subunits among Haemophilus influenzae isolates

Kirk W. McCrea, Jennifer St. Sauver, Carl F. Marrs, Daniel Clemans, Janet R. Gilsdorf

Research output: Contribution to journalArticle

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Abstract

Two proteins, HifD and HifE, have been identified as structural components of Haemophilus influenzae pill. Both are localized at the pilus tip, and Hife appears to mediate pilus adherence to host cells. In this study we examined the immunologic and structural diversity of these pilus subunits among type b H. influenzae (Hib) and nontypeable H. influenzae (NTHI) strains. Western immunoblot analysis revealed that antibodies directed against the C terminus of HifD and Hife from Hib strain Eagan bound to HifD and HifE proteins, respectively, of all piliated Hib and NTHI strains tested. Whole-cell enzyme-linked immunosorbent assays showed that antibodies specific for native HifD or HifE of strain Eagan also bound to all piliated Hib strains but did not bind to the piliated NTHI strains. Antibodies against Hife of strain Eagan inhibited the binding of Hib to human erythrocytes but did not inhibit the binding of NTHI strains. Restriction fragment length polymorphism (RFLP) analysis was used to determine strain-to-strain structural differences within hifD and hifE genes, either by PCR or by nucleotide sequence analysis. DNA and derived amino acid sequence analyses of HifD and HifE confirmed the uniqueness of the RFLP types. The hifD and hire genes of all type b strains showed identical restriction patterns. Analysis ofhifD and hire genes from the NTHI strains, however, revealed seven unique RFLP patterns, suggesting that these genes encode proteins with diverse primary structures. These results indicate that HifD and HifE are immunologically and structurally similar among the Hib strains but vary among the NTHI strains.

Original languageEnglish (US)
Pages (from-to)4788-4796
Number of pages9
JournalInfection and Immunity
Volume66
Issue number10
StatePublished - 1998
Externally publishedYes

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Haemophilus influenzae type b
Haemophilus influenzae
Restriction Fragment Length Polymorphisms
Antibodies
Genes
Proteins
Protein Sequence Analysis
Sequence Analysis
Erythrocytes
Western Blotting
Enzyme-Linked Immunosorbent Assay
Polymerase Chain Reaction
DNA

ASJC Scopus subject areas

  • Immunology

Cite this

Immunologic and structural relationships of the minor pilus subunits among Haemophilus influenzae isolates. / McCrea, Kirk W.; St. Sauver, Jennifer; Marrs, Carl F.; Clemans, Daniel; Gilsdorf, Janet R.

In: Infection and Immunity, Vol. 66, No. 10, 1998, p. 4788-4796.

Research output: Contribution to journalArticle

McCrea, Kirk W. ; St. Sauver, Jennifer ; Marrs, Carl F. ; Clemans, Daniel ; Gilsdorf, Janet R. / Immunologic and structural relationships of the minor pilus subunits among Haemophilus influenzae isolates. In: Infection and Immunity. 1998 ; Vol. 66, No. 10. pp. 4788-4796.
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