The major goal of this proposal is to determine the role of Duffy Antigen/ Receptor for Chemokines (DARC) in endothelial cells during lung inflammation. DARC binds multiple CXC and CC chemokines and is expressed on post-capillary venular endothelial cells, the site where leukocytes traverse the endothelium in response to inflammatory signals released from a site of tissue injury. Although endothelial DARC is upregulated during inflammation, its biological function remains unknown. It's focal location at the site of leukocyte emigration, its upregulation during inflammation, and preservation of expression on endothelial cells in all individuals suggest that it may participate in the regulation of inflammatory cell recruitment at the bloodtissue interface.
The specific aims will explore the following : (1)endothelial DARC's role in chemokine transport, (2) its ability to modify neutrophil transendothelial migration, (3) its in vivo expression in the lung during suppurative pneumonia, (4) and DARC's contribution to neutrophil recruitment in a mouse model of pneumonia. A human endothelial cell line stably expressing DARC cDNA has been developed to study its ability to modify the biological activity of chemokines in vitro. Immunohistochemical analysis of human lungs will define the expression pattern of endothelial DARC during suppurative pneumonia. Finally, DARC's role will be examined in an animal model of bacterial pneumonia using DARC knockout mice. These studies are important, as they will determine endothelial DARC's contribution to the pulmonary recruitment of neutrophils during lung inflammation. Understanding endothelial DARC's function may provide novel strategies in modifying the inflammatory component of specific diseases such as bacterial pneumonia. These studies will further the candidate's research training in cell and molecular biology and provide the foundation for a career as a physicianscientist. The proposed studies will be conducted at the Pulmonary VA Research laboratories under the guidance of the sponsor and co-sponsor. Interactions with consultants throughout the University of Washington research community will enrich the candidate's training and ensure the success of the proposal.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Clinical Investigator Award (CIA) (K08)
Project #
5K08HL070178-04
Application #
6938524
Study Section
Special Emphasis Panel (ZHL1-CSR-M (F2))
Program Officer
Colombini-Hatch, Sandra
Project Start
2002-07-01
Project End
2007-06-30
Budget Start
2005-07-01
Budget End
2006-06-30
Support Year
4
Fiscal Year
2005
Total Cost
$128,493
Indirect Cost
Name
University of Pittsburgh
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
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Lee, Janet S; Rosengart, Matthew R; Kondragunta, Venkateswarlu et al. (2007) Inverse association of plasma IL-13 and inflammatory chemokines with lung function impairment in stable COPD: a cross-sectional cohort study. Respir Res 8:64
Lee, Janet S; Wurfel, Mark M; Matute-Bello, Gustavo et al. (2006) The Duffy antigen modifies systemic and local tissue chemokine responses following lipopolysaccharide stimulation. J Immunol 177:8086-94
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Lee, Janet S; Frevert, Charles W; Matute-Bello, Gustavo et al. (2005) TLR-4 pathway mediates the inflammatory response but not bacterial elimination in E. coli pneumonia. Am J Physiol Lung Cell Mol Physiol 289:L731-8
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Lee, Janet S; Frevert, Charles W; Thorning, David R et al. (2003) Enhanced expression of Duffy antigen in the lungs during suppurative pneumonia. J Histochem Cytochem 51:159-66
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Lee, Janet S; Frevert, Charles W; Wurfel, Mark M et al. (2003) Duffy antigen facilitates movement of chemokine across the endothelium in vitro and promotes neutrophil transmigration in vitro and in vivo. J Immunol 170:5244-51