Expression of Haemophilus ducreyi collagen binding outer membrane protein NcaA is required for virulence in swine and human challenge models of chancroid

Robert A. Fulcher, Leah E. Cole, Diane Janowicz, Kristen L. Toffer, Kate R. Fortney, Barry Katz, Paul E. Orndorff, Stanley Spinola, Thomas H. Kawula

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Haemophilus ducreyi, the etiologic agent of the sexually transmitted genital ulcer disease chancroid, has been shown to associate with dermal collagen fibers within infected skin lesions. Here we describe NcaA, a previously uncharacterized outer membrane protein that is important for H. ducreyi collagen binding and host colonization. An H. ducreyi strain lacking the ncaA gene was impaired in adherence to type I collagen but not fibronectin (plasma or cellular form) or heparin. The mutation had no effect on serum resistance or binding to HaCaT keratinocytes or human foreskin fibroblasts in vitro. Escherichia coli expressing H. ducreyi NcaA bound to type I collagen, demonstrating that NcaA is sufficient to confer collagen attachment. The importance of NcaA in H. ducreyi pathogenesis was assessed using both swine and human experimental models of chancroid. In the swine model, 20% of lesions from sites inoculated with the ncaA mutant were culture positive for H. ducreyi 7 days after inoculation, compared to 73% of wild-type-inoculated sites. The average number of CFU recovered from mutant-inoculated lesions was also significantly reduced compared to that recovered from wild-type-inoculated sites at both 2 and 7 days after inoculation. In the human challenge model, 8 of 30 sites inoculated with wild-type H. ducreyi progressed to the pustular stage, compared to 0 of 30 sites inoculated with the ncaA mutant. Together these results demonstrate that the collagen binding protein NcaA is required for H. ducreyi infection.

Original languageEnglish
Pages (from-to)2651-2658
Number of pages8
JournalInfection and Immunity
Volume74
Issue number5
DOIs
StatePublished - May 2006

Fingerprint

Haemophilus ducreyi
Chancroid
Virulence
Membrane Proteins
Swine
Collagen
Collagen Type I
Haemophilus Infections
Foreskin
Skin
Keratinocytes
Fibronectins
Ulcer
Heparin
Carrier Proteins
Theoretical Models
Fibroblasts
Escherichia coli
Mutation

ASJC Scopus subject areas

  • Immunology

Cite this

Expression of Haemophilus ducreyi collagen binding outer membrane protein NcaA is required for virulence in swine and human challenge models of chancroid. / Fulcher, Robert A.; Cole, Leah E.; Janowicz, Diane; Toffer, Kristen L.; Fortney, Kate R.; Katz, Barry; Orndorff, Paul E.; Spinola, Stanley; Kawula, Thomas H.

In: Infection and Immunity, Vol. 74, No. 5, 05.2006, p. 2651-2658.

Research output: Contribution to journalArticle

Fulcher, Robert A. ; Cole, Leah E. ; Janowicz, Diane ; Toffer, Kristen L. ; Fortney, Kate R. ; Katz, Barry ; Orndorff, Paul E. ; Spinola, Stanley ; Kawula, Thomas H. / Expression of Haemophilus ducreyi collagen binding outer membrane protein NcaA is required for virulence in swine and human challenge models of chancroid. In: Infection and Immunity. 2006 ; Vol. 74, No. 5. pp. 2651-2658.
@article{e9f64fd436f248618420e952a53bd66c,
title = "Expression of Haemophilus ducreyi collagen binding outer membrane protein NcaA is required for virulence in swine and human challenge models of chancroid",
abstract = "Haemophilus ducreyi, the etiologic agent of the sexually transmitted genital ulcer disease chancroid, has been shown to associate with dermal collagen fibers within infected skin lesions. Here we describe NcaA, a previously uncharacterized outer membrane protein that is important for H. ducreyi collagen binding and host colonization. An H. ducreyi strain lacking the ncaA gene was impaired in adherence to type I collagen but not fibronectin (plasma or cellular form) or heparin. The mutation had no effect on serum resistance or binding to HaCaT keratinocytes or human foreskin fibroblasts in vitro. Escherichia coli expressing H. ducreyi NcaA bound to type I collagen, demonstrating that NcaA is sufficient to confer collagen attachment. The importance of NcaA in H. ducreyi pathogenesis was assessed using both swine and human experimental models of chancroid. In the swine model, 20{\%} of lesions from sites inoculated with the ncaA mutant were culture positive for H. ducreyi 7 days after inoculation, compared to 73{\%} of wild-type-inoculated sites. The average number of CFU recovered from mutant-inoculated lesions was also significantly reduced compared to that recovered from wild-type-inoculated sites at both 2 and 7 days after inoculation. In the human challenge model, 8 of 30 sites inoculated with wild-type H. ducreyi progressed to the pustular stage, compared to 0 of 30 sites inoculated with the ncaA mutant. Together these results demonstrate that the collagen binding protein NcaA is required for H. ducreyi infection.",
author = "Fulcher, {Robert A.} and Cole, {Leah E.} and Diane Janowicz and Toffer, {Kristen L.} and Fortney, {Kate R.} and Barry Katz and Orndorff, {Paul E.} and Stanley Spinola and Kawula, {Thomas H.}",
year = "2006",
month = "5",
doi = "10.1128/IAI.74.5.2651-2658.2006",
language = "English",
volume = "74",
pages = "2651--2658",
journal = "Infection and Immunity",
issn = "0019-9567",
publisher = "American Society for Microbiology",
number = "5",

}

TY - JOUR

T1 - Expression of Haemophilus ducreyi collagen binding outer membrane protein NcaA is required for virulence in swine and human challenge models of chancroid

AU - Fulcher, Robert A.

AU - Cole, Leah E.

AU - Janowicz, Diane

AU - Toffer, Kristen L.

AU - Fortney, Kate R.

AU - Katz, Barry

AU - Orndorff, Paul E.

AU - Spinola, Stanley

AU - Kawula, Thomas H.

PY - 2006/5

Y1 - 2006/5

N2 - Haemophilus ducreyi, the etiologic agent of the sexually transmitted genital ulcer disease chancroid, has been shown to associate with dermal collagen fibers within infected skin lesions. Here we describe NcaA, a previously uncharacterized outer membrane protein that is important for H. ducreyi collagen binding and host colonization. An H. ducreyi strain lacking the ncaA gene was impaired in adherence to type I collagen but not fibronectin (plasma or cellular form) or heparin. The mutation had no effect on serum resistance or binding to HaCaT keratinocytes or human foreskin fibroblasts in vitro. Escherichia coli expressing H. ducreyi NcaA bound to type I collagen, demonstrating that NcaA is sufficient to confer collagen attachment. The importance of NcaA in H. ducreyi pathogenesis was assessed using both swine and human experimental models of chancroid. In the swine model, 20% of lesions from sites inoculated with the ncaA mutant were culture positive for H. ducreyi 7 days after inoculation, compared to 73% of wild-type-inoculated sites. The average number of CFU recovered from mutant-inoculated lesions was also significantly reduced compared to that recovered from wild-type-inoculated sites at both 2 and 7 days after inoculation. In the human challenge model, 8 of 30 sites inoculated with wild-type H. ducreyi progressed to the pustular stage, compared to 0 of 30 sites inoculated with the ncaA mutant. Together these results demonstrate that the collagen binding protein NcaA is required for H. ducreyi infection.

AB - Haemophilus ducreyi, the etiologic agent of the sexually transmitted genital ulcer disease chancroid, has been shown to associate with dermal collagen fibers within infected skin lesions. Here we describe NcaA, a previously uncharacterized outer membrane protein that is important for H. ducreyi collagen binding and host colonization. An H. ducreyi strain lacking the ncaA gene was impaired in adherence to type I collagen but not fibronectin (plasma or cellular form) or heparin. The mutation had no effect on serum resistance or binding to HaCaT keratinocytes or human foreskin fibroblasts in vitro. Escherichia coli expressing H. ducreyi NcaA bound to type I collagen, demonstrating that NcaA is sufficient to confer collagen attachment. The importance of NcaA in H. ducreyi pathogenesis was assessed using both swine and human experimental models of chancroid. In the swine model, 20% of lesions from sites inoculated with the ncaA mutant were culture positive for H. ducreyi 7 days after inoculation, compared to 73% of wild-type-inoculated sites. The average number of CFU recovered from mutant-inoculated lesions was also significantly reduced compared to that recovered from wild-type-inoculated sites at both 2 and 7 days after inoculation. In the human challenge model, 8 of 30 sites inoculated with wild-type H. ducreyi progressed to the pustular stage, compared to 0 of 30 sites inoculated with the ncaA mutant. Together these results demonstrate that the collagen binding protein NcaA is required for H. ducreyi infection.

UR - http://www.scopus.com/inward/record.url?scp=33646350248&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33646350248&partnerID=8YFLogxK

U2 - 10.1128/IAI.74.5.2651-2658.2006

DO - 10.1128/IAI.74.5.2651-2658.2006

M3 - Article

C2 - 16622201

AN - SCOPUS:33646350248

VL - 74

SP - 2651

EP - 2658

JO - Infection and Immunity

JF - Infection and Immunity

SN - 0019-9567

IS - 5

ER -