The ability to reproduce endotoxin shock in mice provides a unique opportunity to study the biology of the disease and its consequences. The studies proposed here take advantage of the ease of doing experiments in mice and the newest genetic technology. Dr. Goyert has recently produced mice in which the CD14 gene has been disrupted by homologous recombination. These mice provide a unique opportunity to ask several fundamental questions about the nature of endotoxin shock, particularly as regards CD14- dependent and CD14-independent pathways to endotoxin shock. In addition, this animal model will allow Dr. Goyert to evaluate the relative contribution of endothelial cell responses to sCD14- LPS complexes in endotoxin shock.This is the first time that such a model has been established for delineating the effects of these three mechanisms. She will also continue development of a new animal model that will respond to endotoxin in a CD14-dependent manner through the human CD14 protein. This model will be used to evaluate the efficacy of reagents (anti-human CD14 mAb, recombinant soluble human CD14) on the endotoxin response of mice expressing a human CD14 gene and lacking expression of the murine CD14 gene. The following questions will be addressed: 1. How does the lack of CD14 affect the response of CD14-knockout mice to LPS, Gram-negative bacteria, Gram-positive bacteria, and complexes of LPS and soluble CD14? 2. How effective are soluble human CD14 and anti-human CD14 mAb in preventing endotoxin shock in mice expressing human but not murine CD14? Furthermore, can efficacious, unique CD14-specific mAbs be generated in CD14-deficient mice? 3. What is the three-dimensional structure of CD14? 4. What is the nature of the endothelial cell receptor for the complex of LPS-soluble CD14 and is it present on other cell types that also respond to this complex?

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI023859-13
Application #
2671871
Study Section
Special Emphasis Panel (ZRG5-BM-1 (04))
Project Start
1989-01-01
Project End
2000-03-31
Budget Start
1998-04-01
Budget End
1999-03-31
Support Year
13
Fiscal Year
1998
Total Cost
Indirect Cost
Name
North Shore University Hospital
Department
Type
DUNS #
City
Manhasset
State
NY
Country
United States
Zip Code
11030
Killoran, Kristin E; Miller, Amber D; Uray, Karen S et al. (2014) Role of innate immunity and altered intestinal motility in LPS- and MnCl2-induced intestinal intussusception in mice. Am J Physiol Gastrointest Liver Physiol 306:G445-53
Brinkworth, J F; Pechenkina, E A; Silver, J et al. (2012) Innate immune responses to TLR2 and TLR4 agonists differ between baboons, chimpanzees and humans. J Med Primatol 41:388-93
Metkar, Shalaka; Kim, Kwang Sik; Silver, Jack et al. (2012) Differential expression of CD14-dependent and independent pathways for chemokine induction regulates neutrophil trafficking in infection. J Leukoc Biol 92:389-96
Metkar, Shalaka; Awasthi, Shanjana; Denamur, Erick et al. (2007) Role of CD14 in responses to clinical isolates of Escherichia coli: effects of K1 capsule expression. Infect Immun 75:5415-24
Chen, Guoqian; Li, Jianhua; Ochani, Mahendar et al. (2004) Bacterial endotoxin stimulates macrophages to release HMGB1 partly through CD14- and TNF-dependent mechanisms. J Leukoc Biol 76:994-1001
Gangloff, S C; Hijiya, N; Haziot, A et al. (1999) Lipopolysaccharide structure influences the macrophage response via CD14-independent and CD14-dependent pathways. Clin Infect Dis 28:491-6
Arditi, M; Zhou, J; Dorio, R et al. (1993) Endotoxin-mediated endothelial cell injury and activation: role of soluble CD14. Infect Immun 61:3149-56
Ferrero, E; Jiao, D; Tsuberi, B Z et al. (1993) Transgenic mice expressing human CD14 are hypersensitive to lipopolysaccharide. Proc Natl Acad Sci U S A 90:2380-4
Chang, M D; Pollard, J W; Khalili, H et al. (1993) Mouse placental macrophages have a decreased ability to present antigen. Proc Natl Acad Sci U S A 90:462-6
Haziot, A; Tsuberi, B Z; Goyert, S M (1993) Neutrophil CD14: biochemical properties and role in the secretion of tumor necrosis factor-alpha in response to lipopolysaccharide. J Immunol 150:5556-65

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