Haemophilus ducreyi is the etiologic agent of chancroid, a sexually transmitted genital ulcer disease. Verylittle Is known about how this unencapsulated. Gram-negative bacterium evades host defenses and causesdemrial lesion development. However, it has been shown that H. ducreyi has the ability to preventphagocytosis of both itself and secondary targets by macrophages in vitro. We identified two extremely largeH. ducreyi proteins, designated LspA1 and LspA2, that are released into H. ducreyi culture supernatant fluidand which are necessary for the observed inhibition of phagocytic activity. That these proteins are relevantto disease production was proven by the finding that a H. ducreyi mutant unable to express either LspA1 orLspA2 had drastically reduced virulence in both animal and human models of experimental chancroid. Littleis known, however, about how the LspA proteins inhibit phagocytic activity. The proposed research projectin this MERIT extension application will investigate the stoicture, function, and expression of the LspAproteins. In the first Specific Aim, we will capitalize on our recent successful cloning of the H. ducreyiIspAl gene to purify a functional recombinant LspAl protein and then determine the composition of theactive form of this protein. In the second Specific Aim, we will elucidate the mechanism of action involvedin the inhibition of phagocytic activity by the LspA proteins. We already have data which indicate that theLspA proteins can affect signaling pathways that control phagocytosis in macrophages, and our researchefforts will be focused on the most proximal elements, and especially the Src family protein tyrosine kinases,in the phagocytic signaling cascade. In the third Specific Aim, we will characterize the H. ducreyi CpxRAtwo-component signal transduction system that is responsible for control of expression of both the LspAproteins as well as other co-regulated gene products of this bacterium.

Public Health Relevance

The relevance of this research to public health involves the new infomnation that will be gained about H.ducreyi inhibits phagocytosis, one of the primary defense mechanisms of the human body. The ability ofphagocytes to engulf and kill bacteria is essential to preventing or curing infectious diseases. Infomnationgained from this study will help us understand how phagocytes control this protective activity.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
4R37AI032011-19
Application #
7906548
Study Section
Special Emphasis Panel (NSS)
Program Officer
Hiltke, Thomas J
Project Start
1992-01-01
Project End
2016-01-31
Budget Start
2011-02-25
Budget End
2012-01-31
Support Year
19
Fiscal Year
2011
Total Cost
$396,250
Indirect Cost
Name
University of Texas Sw Medical Center Dallas
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
800771545
City
Dallas
State
TX
Country
United States
Zip Code
75390
Gangaiah, Dharanesh; Labandeira-Rey, Maria; Zhang, Xinjun et al. (2014) Haemophilus ducreyi Hfq contributes to virulence gene regulation as cells enter stationary phase. MBio 5:e01081-13
Dodd, Dana A; Worth, Randall G; Rosen, Michael K et al. (2014) The Haemophilus ducreyi LspA1 protein inhibits phagocytosis by using a new mechanism involving activation of C-terminal Src kinase. MBio 5:e01178-14
Labandeira-Rey, Maria; Dodd, Dana A; Brautigam, Chad A et al. (2013) The Haemophilus ducreyi Fis protein is involved in controlling expression of the lspB-lspA2 operon and other virulence factors. Infect Immun 81:4160-70
Labandeira-Rey, Maria; Dodd, Dana; Fortney, Kate R et al. (2011) A Haemophilus ducreyi CpxR deletion mutant is virulent in human volunteers. J Infect Dis 203:1859-65
Labandeira-Rey, Maria; Brautigam, Chad A; Hansen, Eric J (2010) Characterization of the CpxRA regulon in Haemophilus ducreyi. Infect Immun 78:4779-91
Labandeira-Rey, Maria; Mock, Jason R; Hansen, Eric J (2009) Regulation of expression of the Haemophilus ducreyi LspB and LspA2 proteins by CpxR. Infect Immun 77:3402-11
Labandeira-Rey, Maria; Janowicz, Diane M; Blick, Robert J et al. (2009) Inactivation of the Haemophilus ducreyi luxS gene affects the virulence of this pathogen in human subjects. J Infect Dis 200:409-16