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Am J Physiol Cell Physiol (October 26, 2005). doi:10.1152/ajpcell.00166.2005
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Submitted on April 8, 2005
Accepted on October 18, 2005

Innate Immune Responses of Human Tracheal Epithelium to P. aeruginosa Flagellin, TNF{alpha} and IL1{beta}

Jill Tseng1, Jiun Do1, Jonathan H Widdicombe2, and Terry E Machen1*

1 Molecular and Cell Biology, University of California at Berkeleyifornia - Berkeley, Berkeley, CA, USA
2 Human Physiology, University of California at Davis, Davis, CA, USA

* To whom correspondence should be addressed. E-mail: tmachen{at}calmail.berkeley.edu.

We measured innate immune responses by primary human tracheal epithelial cells grown as confluent, pseudostratified layers during exposure to inflammatory activators on apical vs basolateral surfaces. Apical P. aeruginosa strain PAK (but not isogenic flagellin mutant PAK{Delta}fliC), flagellin and flagellin+PAK{Delta}fliC activated NF-{kappa}B and IL8 expression and secretion. LPS (5x10-8 g/ml, approximately 2x10-8 M) elicited only 1.2-fold increases in NF-{kappa}B activation and IL8 secretion, while flagellin (10-6 g/ml, approximately 2.5x10-8 M) elicited 4-7-fold increases in NF-{kappa}B activation and IL8 secretion, and there was little difference in effects of flagellin vs flagellin + LPS. Thus, HTE cells were insensitive to LPS compared to flagellin. Flagellin activated NF-{kappa}B in columnar but not basal cells. IL1{beta}+TNF{alpha} elicited similar responses. Basolateral flagellin or IL1{beta}+TNF{alpha} caused 1.5- to 4-fold larger responses, consistent with the fact that NF-{kappa}B activation occurred in both columnar and basal cells. MyD88 (toll receptor-associated adapter), IL1 receptor (IL1R1) and TNF receptor (TNFR1) were expressed in columnar and basal cells. ZO-1 was localized to apical, tight junctional regions of columnar cells but not to basal cells. We infer the following from these data: (i) Flagellin is necessary and sufficient to trigger inflammatory responses in columnar cells during accumulation of P. aeruginosa in the airway surface liquid (ASL); columnar cells express TLR5 and MyD88, often associated with flagellin-activated cell signaling; (ii) IL1{beta}+TNF{alpha} in the ASL also activate columnar cells, and these cells also express IL1R1 and TNFR1. (iii) Apical flagellin, IL1{beta} or TNF{alpha} do not activate basal cells because tight junctions between columnar cells prevent access from the apical surface to the basal cells. (iv) Exposure of basolateral surfaces to inflammatory activators elicits larger responses because both columnar and basal cells are activated, likely because both cell types express receptors for flagellin, IL1{beta} and TNF{alpha}.




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