Anaplasma phagocytophilum is an obligate intracellular bacterial pathogen that invades neutrophils and endothelial cells to cause the emerging and potentially fatal infection, human granulocytic anaplasmosis (HGA). A. phagocytophilum converts its host neutrophil into a Trojan Horse that facilitates pathogen replication and dissemination. It downregulates the neutrophil antimicrobial response, thereby raising susceptibility to opportunistic infections. Thus, blocking A. phagocytophilum infection of neutrophils would potentially prevent the stage of HGA associated with pathogen dissemination and increased risk of opportunistic infections. A. phagocytophilum infects microvascular endothelial cells of heart and liver. Endothelial cells are also implicated as the initial cell typ that A. phagocytophilum infects following inoculation via tick feeding. Infected endothelial cells are capable of transferring the bacterium to neutrophils. Therefore, abrogating A. phagocytophilum invasion of endothelial cells would potentially prevent infection of major organs and initial establishment of infection. The overall goal of this project is to identify and functioally characterize A. phagocytophilum outer membrane proteins that facilitate infection. We have identified outer membrane protein A (OmpA) and Asp14 (14-kDa Ap surface protein) as the first two A. phagocytophilum proteins that are critical for invasion of both myeloid cells and endothelial cells.
In Aim 1, we will identify the OmpA and Asp14 domains that mediate A. phagocytophilum uptake and determine if either protein is sufficient for invasion.
In Aim 2, we will identify the host cell receptors of OmpA and Asp14.
In Aim 3, we will directly assess the relevance of OmpA and Asp14 to infectivity using novel transgenic A. phagocytophilum organisms that can be induced to express antisense RNA against ompA or asp14 to knock down OmpA or Asp14 expression. Achieving our goals will provide a robust understanding of A. phagocytophilum cellular invasion and identify the first two invasin-receptor pairs for any Anaplasmataceae pathogen. It will also yield a valuable tool for assessing gene function in obligate intracellular bacteria. Lastly, our work will potentially aid the design of novel intervenion strategies that target OmpA and Asp14 to prevent HGA by blocking multiple stages of infection.

Public Health Relevance

Human granulocytic anaplasmosis (HGA) is an emerging, potentially fatal disease caused by a bacterium that invades blood vessel endothelial cells and white blood cells called neutrophils. We have identified two bacterial factors that are critical fr infection of both host cell types. The proposed research will determine how the two bacterial factors mediate infection. Doing so will aid the design of novel strategies for preventing HGA.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
High Priority, Short Term Project Award (R56)
Project #
2R56AI072683-07
Application #
8510769
Study Section
Host Interactions with Bacterial Pathogens Study Section (HIBP)
Program Officer
Mukhopadhyay, Suman
Project Start
2012-07-24
Project End
2013-06-30
Budget Start
2012-07-24
Budget End
2013-06-30
Support Year
7
Fiscal Year
2012
Total Cost
$367,810
Indirect Cost
$117,810
Name
Virginia Commonwealth University
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
105300446
City
Richmond
State
VA
Country
United States
Zip Code
23298
Truchan, Hilary K; VieBrock, Lauren; Cockburn, Chelsea L et al. (2016) Anaplasma phagocytophilum Rab10-dependent parasitism of the trans-Golgi network is critical for completion of the infection cycle. Cell Microbiol 18:260-81
Truchan, Hilary K; Seidman, David; Carlyon, Jason A (2013) Breaking in and grabbing a meal: Anaplasma phagocytophilum cellular invasion, nutrient acquisition, and promising tools for their study. Microbes Infect 15:1017-25
Huang, Bernice; Hubber, Andree; McDonough, Justin A et al. (2010) The Anaplasma phagocytophilum-occupied vacuole selectively recruits Rab-GTPases that are predominantly associated with recycling endosomes. Cell Microbiol 12:1292-307