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Proteomic analysis of the airway surface liquid: modulation by proinflammatory cytokines

¹Laboratory of Uremic Physiopathology and ²Laboratory of Molecular Genetics, Istituto Giannina Gaslini, Genoa; ³Laboratoty of Functional Proteomics, Molecular Biology Department, Università degli Studi di Siena, Siena; and ⁴Renal Child Foundation, Genoa, Italy

Submitted 9 March 2006 ; accepted in final form 21 September 2006

The airway surface is covered by a fluid, the airway surface liquid, interposed between the mucous layer and the epithelium. The airway surface liquid contains proteins, secreted by different cell types, that may have pro-/anti-inflammatory or bactericidal functions or have a role in the mucociliary clearance. We have used a proteomics approach to identify the proteins secreted by an isolated in vitro model of human airway epithelium, at resting and under proinflammatory conditions, as a strategy to define the factors involved in epithelial barrier function. To this aim, we have analyzed the airway surface liquid from human bronchial epithelial cells grown as polarized monolayers in the presence and absence of inflammatory stimuli such as IL-4, IL-1, TNF-, and IFN-. Two-dimensional electrophoresis followed by mass spectrometry analysis has allowed the identification of 175 secreted protein spots, among which are immune-related proteins, structural proteins, an actin severer, some protease inhibitors, and a metalloproteinase. Comparisons between treated and untreated conditions have shown that the expression of several proteins was significantly modified by the different cytokines. Our results indicate that the surface epithelium is an active player in the epithelial barrier function and that inflammatory conditions may modulate protein secretion.

human bronchial epithelial cells; interleukin-4; interleukin-1; tumor necrosis factor-; interferon-

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