Haemophilus ducreyi is the etiologic agent of chancroid, a sexually transmitted genital ulcer disease. Very little is known about how this unencapsulated, Gram-negative bacterium evades host defenses and causes dermal lesion development. However, it has been shown that H. ducreyi has the ability to resist phagocytosis in vivo. Moreover, this bacterium can prevent phagocytosis of both itself and secondary targets by macrophages in vitro. We have identified two extremely large H. ducreyi proteins, designated LspAt and LspA2, that are released into H. ducreyi culture supernatant fluid and which are responsible for the observed inhibition of phagocytic activity. That these proteins are relevant to disease production was proven by the finding that a H. ducreyi mutant unable to express either LspA1 or LspA2 had drastically reduced virulence in both animal and human models of experimental chancroid. Nothing is known, however, about how the LspA proteins inhibit phagocytic activity. The proposed research project is focused on the structure, function, and expression of the LspA proteins. In the first Specific Aim, we will purify either a functional LspAt fusion protein or native LspA1 protein and determine the composition of the active form of this protein. In the second Specific Aim,we will elucidate the mechanism of action involved in the inhibition of phagocytic activity by the LspA proteins. We already have data which indicate that the LspA proteins can affect signaling pathways that control phagocytosis in macrophages. In the third Specific Aim, we will identify the H. ducreyi gene products that are responsible for control of expression of the LspA proteins by this bacterium. The relevance of this research to public health involves the new information that will be gained about phagocytosis, one of the primary defense mechanisms of the human body. The ability of phagocytes to engulf and kill bacteria is essential to preventing or curing infectious diseases. Information gained 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 #
5R37AI032011-18
Application #
7753629
Study Section
Bacterial Pathogenesis Study Section (BACP)
Program Officer
Hiltke, Thomas J
Project Start
1992-01-01
Project End
2011-01-31
Budget Start
2010-02-01
Budget End
2011-01-31
Support Year
18
Fiscal Year
2010
Total Cost
$370,138
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
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
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
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; 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
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