Staphylococcus aureus is well recognized as a virulent pathogen, which continues to cause life-threatening disease. Its invasive potential is clearly demonstrated by its ability to colonize and infect endovascular tissue. Despite its continued frequency and importance, little is known about the pathogenesis of this endovascular infection. Our laboratory has developed an in vitro assay to explore the dynamics of staphylococcal - human endothelial cell interactions. Recently, we have isolated an endothelial cell membrane protein which binds staphylococci. In the present proposal, we plan to explore the role of this protein in the initial binding interaction and the consequences of staphylococcal infection to the endothelial cell. We hypothesize that invasive staphylococcal disease is the result of specific S. aureus adhesin - endothelial cell receptor mechanisms. Injury to the cell and alterations of the cell surface following infection, may help explain some of the devastating clinical complications encountered in patients with this disease.
Our specific aims i ncluding the following: 1. What are the characteristics of the isolated endothelial cell membrane binding protein? The protein will be further purified and characterized biochemically. Kinetic binding studies with iodinated protein will further elucidate the specificity and affinity of binding to different S. aureus isolates. The protein's effect on S. aureus adherence to endothelial cells in the infection assay will be determined. Polyclonal antibodies, prepared against the purified protein, will be used in immnofluorescent and ultrastructural studies to define the topography of the binding protein on the surface of endothelial cells harvested from different sources. 2. What are the consequences of staphylococcal infection of the endothelial cell? Injury to the cells by staphylococci may result in the modulation of the surface with increased expression of the binding protein, Fc receptors, tissue factor or increase platelet or white blood cell adhesivity which may contribute to the progression of the disease. The influence of infection on these factors will be determined. 3. Are there additional factors for S. aureus on the endothelial cell surface which can be identified? Monoclonal antibodies directed against human endothelial cells will be screened for their ability to block S. aureus adherence to these cells. These antibodies will then be employed to identify, isolate and characterize the receptors.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
2R01HL034171-04
Application #
3346846
Study Section
Bacteriology and Mycology Subcommittee 1 (BM)
Project Start
1985-04-01
Project End
1991-03-31
Budget Start
1988-04-01
Budget End
1989-03-31
Support Year
4
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Montefiore Medical Center (Bronx, NY)
Department
Type
DUNS #
City
New York
State
NY
Country
United States
Zip Code
10467
Tompkins, D C; Blackwell, L J; Hatcher, V B et al. (1992) Staphylococcus aureus proteins that bind to human endothelial cells. Infect Immun 60:965-9
Elliott, D A; Hatcher, V B; Lowy, F D (1991) A 220-kilodalton glycoprotein in yeast extract inhibits Staphylococcus aureus adherence to human endothelial cells. Infect Immun 59:2222-3
Tsai, H M; Nagel, R L; Hatcher, V B et al. (1991) The high molecular weight form of endothelial cell von Willebrand factor is released by the regulated pathway. Br J Haematol 79:239-45
Tompkins, D C; Hatcher, V B; Patel, D et al. (1990) A human endothelial cell membrane protein that binds Staphylococcus aureus in vitro. J Clin Invest 85:1248-54
Bengualid, V; Hatcher, V B; Diamond, B et al. (1990) Staphylococcus aureus infection of human endothelial cells potentiates Fc receptor expression. J Immunol 145:4279-83
Blumberg, E A; Hatcher, V B; Lowy, F D (1988) Acidic fibroblast growth factor modulates Staphylococcus aureus adherence to human endothelial cells. Infect Immun 56:1470-4
Lowy, F D; Fant, J; Higgins, L L et al. (1988) Staphylococcus aureus--human endothelial cell interactions. J Ultrastruct Mol Struct Res 98:137-46
Kessler, J A; Conn, G; Hatcher, V B (1986) Isolated plasma membranes regulate neurotransmitter expression and facilitate effects of a soluble brain cholinergic factor. Proc Natl Acad Sci U S A 83:3528-32
Gimenez-Gallego, G; Conn, G; Hatcher, V B et al. (1986) The complete amino acid sequence of human brain-derived acidic fibroblast growth factor. Biochem Biophys Res Commun 138:611-7
Gordon, P B; Zanger, D R; Hatcher, V B (1986) Extracellular matrix proteoglycans and cell-substratum adhesion of human endothelial cells: the effect of methyl beta-D-xylopyranoside. Carbohydr Res 151:121-34

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