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Published ahead of print on June 10, 2004, doi:10.1165/rcmb.2003-0388OC

Am. J. Respir. Cell Mol. Biol., Volume 31, Number 3, September 2004, 358-364

A more recent version of this article appeared on September 1, 2004
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Submitted on October 30, 2003
Revised on June 8, 2004

Activation of Airway Epithelial Cells by Toll-like Receptor Agonists

Quan Sha1, Ai Q Truong-Tran1, James R Plitt1, Lisa A Beck1, and Robert P Schleimer1*

1 Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USA

* To whom correspondence should be addressed. E-mail: rschleim{at}jhmi.edu.

Toll-like receptors (TLR) play an important role in pathogen recognition and innate immunity. We investigated the presence and function of TLRs in the BEAS-2B airway epithelial cell line and primary bronchial epithelial cells. Standard RT-PCR analysis and Taqman real-time PCR revealed that BEAS-2B cells express mRNA for TLR 1-10. Several TLR ligands were tested for their ability to activate gene expression in BEAS-2B cells using limited microarray analyses focusing on genes of the chemokine and chemokine receptor family, cytokines and signaling pathways. While the TLR3 ligand dsRNA was the most effective epithelial activator, clear responses to flagellin, LPS, CpG, PGN and zymosan were also observed. Real-time PCR and/or ELISA were used to confirm results obtained with microarrays for five of the induced genes: IL-8, serum amyloid A (SAA), TLR3, MIP-3{alpha} and GM-CSF. Stimulation of epithelial cells with dsRNA induced levels of IL-8 exceeding 20 ng/ml and levels of SAA exceeding 80 ng/ml. Double stranded RNA, LPS, zymosan A and flagellin also induced expression of MIP-3{alpha} and GM-CSF which may facilitate immature dendritic cell migration and maturation. These results suggest that airway epithelial cells express several TLRs and that they are functionally active. Epithelial expression of TLRs may be of importance in inflammation and immunity in the airways in response to inhaled pathogens.




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