Eosinophils play a key role in the pathogenesis of respiratory syncytial virus (RSV)- induced airway inflammation and epithelial damage, which may predispose to allergic sensitization and development of asthma. We have shown that: 1) RSV infection of airway epithelial cells induces adhesion-dependent, degranulation of eosinophils; 2) The integral membrane glycoprotein, polymeric Ig receptor (pIgR), is up-regulated on RSV-infected airway epithelial cells and, in its secreted form (secretory component, SC), is a potent stimulus for eosinophil degranulation; 3) RSV is able to infect directly human eosinophils, which induces MAP kinase activity, activation of the potent transcription factor NF-kappaB, and production of chemokines. We hypothesize that pIgR expression on epithelial cells, activation of the Raf-1-MEK-MAP kinase pathway and nuclear translocation of NF-kappaB are critical events in the eosinophil activation during RSV infection. The following specific aims are proposed: 1) To characterize the role of pIgR in eosinophil degranulation induced by RSV-infected airway epithelial. The role pIgR in RSV- induced eosinophil degranulation will be studied by blocking the process using a panel of neutralizing monoclonal antibodies that recognized different epitopes of the pIgR, and by co-culture assays of eosinophils with epithelial cells stably transfected with the human pIgR. 2) To investigate the activation of NF-kappaB in RSV- infected human eosinophils and to determine its role in chemokine gene expression. The NF-kappaB family members that are activated by eosinophils by RSV and the mechanisms of NF-kappaB antisense oligonucleotides or double- stranded competitor oligonucleotides will be employed to determine their effect on RANTES and IL-8 secretion. 3) To study the Raf-1-MEK-MAP kinase pathway in RSV-infected eosinophils and to assess its importance in NF-kappaB activation and chemokine production. The mechanisms of Raf-1 activation in RSV-infected eosinophils, will be studied by examining the involvement of tyrosine kinases, protein kinase C, and p21 ras molecules in the activation of Raf-1 kinase. We will test the hypothesis that NF-kappaB activation and chemokine production (RANTES and IL-8) in RSV-stimulated eosinophils are linked to the activation of the Raf-1-MEK-MAP kinase pathway by the use of specific kinase inhibitors, antisense oligonucleotides, and dominant-negative mutants. Identification of critical molecules involved in eosinophil functions will help in identifying specific inhibitors for treatment of acute RSV infection and for prevention of its sequelae, including asthma.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program Projects (P01)
Project #
5P01AI046004-02
Application #
6344675
Study Section
Special Emphasis Panel (ZAI1)
Project Start
2000-09-01
Project End
2001-08-31
Budget Start
Budget End
Support Year
2
Fiscal Year
2000
Total Cost
$148,391
Indirect Cost
Name
University of Texas Medical Br Galveston
Department
Type
DUNS #
041367053
City
Galveston
State
TX
Country
United States
Zip Code
77555
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Leonard, Patricia; Sur, Sanjiv (2003) Interleukin-12: potential role in asthma therapy. BioDrugs 17:1-7

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