Human β-defensin-3 alters, but does not inhibit, the binding of Porphyromonas gingivalis haemagglutinin B to the surface of human dendritic cells

Jonathan R. Van Hemert, Erica N. Recker, Deborah Dietrich, Ann Progulske-Fox, Zoya Bronislavovna Kurago, Katherine S. Walters, Joseph E. Cavanaugh, Kim A. Brogden

Research output: Contribution to journalArticle

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Abstract

Human β-defensin-3 (HBD3) is a small, cationic, host defence peptide with broad antimicrobial activities and diverse innate immune functions. HBD3 binds to many microbial antigens and, in this study, we hypothesised that the known binding of HBD3 to Porphyromonas gingivalis recombinant haemagglutinin B (rHagB) alters, but does not inhibit, the binding of rHagB to human dendritic cells. To test this, human myeloid dendritic cells were incubated for 5 min with rHagB, HBD3 + rHagB (10:1 molar ratio), HBD3 or 0.1 M phosphate-buffered saline (PBS) (pH 7.2) and were then rapidly fixed and processed for confocal microscopy and ultramicrotomy. rHagB and HBD3 could be detected with primary monoclonal mouse antibody to rHagB (MoAb 1858) or polyclonal rabbit antibody to HBD3 (P241) and secondary fluorescent-labelled anti-mouse or anti-rabbit antibodies (confocal microscopy) or protein A-colloidal gold (immunoelectron microscopy). In cells incubated with rHagB only, fluorescence and protein A-colloidal gold were seen at the cell surface and throughout the cytoplasm. In cells incubated with HBD3 + rHagB, fluorescence was observed only at the cell surface in a 'string of pearls' configuration. Overall, these results suggest that HBD3 binding to rHagB alters, but does not inhibit, the binding of rHagB to human myeloid dendritic cells.

Original languageEnglish (US)
Pages (from-to)75-79
Number of pages5
JournalInternational Journal of Antimicrobial Agents
Volume40
Issue number1
DOIs
StatePublished - Jul 1 2012
Externally publishedYes

Fingerprint

Defensins
Hemagglutinins
Dendritic Cells
Gold Colloid
Staphylococcal Protein A
Myeloid Cells
Confocal Microscopy
Fluorescence
Microtomy
Rabbits
Porphyromonas gingivalis HagB protein
Immunoelectron Microscopy
Anti-Idiotypic Antibodies
Cytoplasm
Phosphates
Monoclonal Antibodies
Antigens
Peptides

Keywords

  • Confocal microscopy
  • Defensins
  • Dendritic cells
  • HBD3
  • Haemagglutinin B
  • Human β-defensin-3
  • Porphyromonas gingivalis

ASJC Scopus subject areas

  • Microbiology (medical)
  • Infectious Diseases
  • Pharmacology (medical)

Cite this

Human β-defensin-3 alters, but does not inhibit, the binding of Porphyromonas gingivalis haemagglutinin B to the surface of human dendritic cells. / Van Hemert, Jonathan R.; Recker, Erica N.; Dietrich, Deborah; Progulske-Fox, Ann; Kurago, Zoya Bronislavovna; Walters, Katherine S.; Cavanaugh, Joseph E.; Brogden, Kim A.

In: International Journal of Antimicrobial Agents, Vol. 40, No. 1, 01.07.2012, p. 75-79.

Research output: Contribution to journalArticle

Van Hemert, Jonathan R. ; Recker, Erica N. ; Dietrich, Deborah ; Progulske-Fox, Ann ; Kurago, Zoya Bronislavovna ; Walters, Katherine S. ; Cavanaugh, Joseph E. ; Brogden, Kim A. / Human β-defensin-3 alters, but does not inhibit, the binding of Porphyromonas gingivalis haemagglutinin B to the surface of human dendritic cells. In: International Journal of Antimicrobial Agents. 2012 ; Vol. 40, No. 1. pp. 75-79.
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abstract = "Human β-defensin-3 (HBD3) is a small, cationic, host defence peptide with broad antimicrobial activities and diverse innate immune functions. HBD3 binds to many microbial antigens and, in this study, we hypothesised that the known binding of HBD3 to Porphyromonas gingivalis recombinant haemagglutinin B (rHagB) alters, but does not inhibit, the binding of rHagB to human dendritic cells. To test this, human myeloid dendritic cells were incubated for 5 min with rHagB, HBD3 + rHagB (10:1 molar ratio), HBD3 or 0.1 M phosphate-buffered saline (PBS) (pH 7.2) and were then rapidly fixed and processed for confocal microscopy and ultramicrotomy. rHagB and HBD3 could be detected with primary monoclonal mouse antibody to rHagB (MoAb 1858) or polyclonal rabbit antibody to HBD3 (P241) and secondary fluorescent-labelled anti-mouse or anti-rabbit antibodies (confocal microscopy) or protein A-colloidal gold (immunoelectron microscopy). In cells incubated with rHagB only, fluorescence and protein A-colloidal gold were seen at the cell surface and throughout the cytoplasm. In cells incubated with HBD3 + rHagB, fluorescence was observed only at the cell surface in a 'string of pearls' configuration. Overall, these results suggest that HBD3 binding to rHagB alters, but does not inhibit, the binding of rHagB to human myeloid dendritic cells.",
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AU - Recker, Erica N.

AU - Dietrich, Deborah

AU - Progulske-Fox, Ann

AU - Kurago, Zoya Bronislavovna

AU - Walters, Katherine S.

AU - Cavanaugh, Joseph E.

AU - Brogden, Kim A.

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N2 - Human β-defensin-3 (HBD3) is a small, cationic, host defence peptide with broad antimicrobial activities and diverse innate immune functions. HBD3 binds to many microbial antigens and, in this study, we hypothesised that the known binding of HBD3 to Porphyromonas gingivalis recombinant haemagglutinin B (rHagB) alters, but does not inhibit, the binding of rHagB to human dendritic cells. To test this, human myeloid dendritic cells were incubated for 5 min with rHagB, HBD3 + rHagB (10:1 molar ratio), HBD3 or 0.1 M phosphate-buffered saline (PBS) (pH 7.2) and were then rapidly fixed and processed for confocal microscopy and ultramicrotomy. rHagB and HBD3 could be detected with primary monoclonal mouse antibody to rHagB (MoAb 1858) or polyclonal rabbit antibody to HBD3 (P241) and secondary fluorescent-labelled anti-mouse or anti-rabbit antibodies (confocal microscopy) or protein A-colloidal gold (immunoelectron microscopy). In cells incubated with rHagB only, fluorescence and protein A-colloidal gold were seen at the cell surface and throughout the cytoplasm. In cells incubated with HBD3 + rHagB, fluorescence was observed only at the cell surface in a 'string of pearls' configuration. Overall, these results suggest that HBD3 binding to rHagB alters, but does not inhibit, the binding of rHagB to human myeloid dendritic cells.

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