Haemophilus ducreyi causes the genital ulcer disease, chancroid. Chancroid facilitates the acquisition and transmission of HIV. Understanding bacterial factors that contribute to the pathogenesis of H. ducreyi infection is a prerequisite for the development of strategies to prevent chancroid. The Spinola laboratory developed a human infection model that mimics the papular and pustular stages of natural chancroid. As a fellow, I conducted several human challenge trials which prompted my interest in bacterial pathogenesis. The Spinola laboratory used the technique of SCOTS (Selective Capture of Transcribed Sequences) on cDNA made from experimental pustules of human volunteers to identify transcripts of H. ducreyi that may be upregulated in vivo. Eleven of the transcripts identified in SCOTS corresponded to lipoproteins. Six of these (HD0192, HD1170, HD1589, HD1629, HD1808 and HD1829) may localize to the outer membrane based on their predicted structures, encode proteins predicted to be >10kDa (and therefore likely to display surface epitopes), were conserved among 10 H. ducreyi strains of different origins, and had transcripts detected in additional infected samples. Two of the lipoprotein transcripts were also upregulated during infection based on microarray data analysis. We hypothesize that these 6 lipoproteins, which are expressed in vivo, are important in H. ducreyi pathogenesis and that they serve as immune targets that may induce bactericidal or opsonophagocytic antibodies. To test these hypotheses, our specific aims include: to confirm by quantitative RT-PCR whether the lipoprotein transcripts identified in SCOTS are upregulated or constitutively expressed in pustules;to examine whether isogenic mutants in upregulated or constitutively expressed lipoprotein genes are attenuated for pustule formation;to characterize the lipoproteins as immune targets. An important feature of this K08 application is a formal didactic component in microbiology and immunology. Lay summary: H. ducreyi is a germ that causes genital ulcers. In order to cause disease, H. ducreyi requires certain proteins. We seek to answer two questions: Which lipoproteins are important in H. ducreyi's ability to cause disease and can any of them serve as vaccines?

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Clinical Investigator Award (CIA) (K08)
Project #
5K08AI074657-03
Application #
7648155
Study Section
Microbiology and Infectious Diseases B Subcommittee (MID)
Program Officer
Hiltke, Thomas J
Project Start
2007-07-05
Project End
2012-05-31
Budget Start
2009-06-01
Budget End
2010-05-31
Support Year
3
Fiscal Year
2009
Total Cost
$118,869
Indirect Cost
Name
Indiana University-Purdue University at Indianapolis
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
603007902
City
Indianapolis
State
IN
Country
United States
Zip Code
46202
Janowicz, Diane M; Zwickl, Beth W; Fortney, Kate R et al. (2014) Outer membrane protein P4 is not required for virulence in the human challenge model of Haemophilus ducreyi infection. BMC Microbiol 14:166
Janowicz, Diane M; Cooney, Sean A; Walsh, Jessica et al. (2011) Expression of the Flp proteins by Haemophilus ducreyi is necessary for virulence in human volunteers. BMC Microbiol 11:208
Janowicz, Diane M; Li, Wei; Bauer, Margaret E (2010) Host-pathogen interplay of Haemophilus ducreyi. Curr Opin Infect Dis 23:64-9
Bauer, Margaret E; Townsend, Carisa A; Doster, Ryan S et al. (2009) A fibrinogen-binding lipoprotein contributes to the virulence of Haemophilus ducreyi in humans. J Infect Dis 199:684-92
Janowicz, Diane M; Ofner, Susan; Katz, Barry P et al. (2009) Experimental infection of human volunteers with Haemophilus ducreyi: fifteen years of clinical data and experience. J Infect Dis 199:1671-9
Bauer, Margaret E; Fortney, Kate R; Harrison, Alistair et al. (2008) Identification of Haemophilus ducreyi genes expressed during human infection. Microbiology 154:1152-60