Title

Effects of CFTR, interleukin-10, and Pseudomonas aeruginosa on gene expression profiles in a CF bronchial epithelial cell Line

UMMS Affiliation

Department of Pediatrics; Gene Therapy Center

Date

9-1-2004

Document Type

Article

Medical Subject Headings

Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Gene Expression Profiling; Interleukin-10; Pseudomonas aeruginosa

Disciplines

Allergy and Immunology | Pediatrics | Respiratory Tract Diseases

Abstract

Mutations in CFTR lead to a complex phenotype that includes increased susceptibility to Pseudomonas infections, a functional deficiency of IL-10, and an exaggerated proinflammatory cytokine response. We examined the effects of CFTR gene correction on the gene expression profile of a CF bronchial epithelial cell line (IB3-1) and determined which CF-related gene expression changes could be reversed by IL-10 expression. We performed microarray experiments to monitor the gene expression profile of three cell lines over a time course of exposure to Pseudomonas. At baseline, we identified 843 genes with statistically different levels of expression in CFTR-corrected (S9) cells compared to the IB3-1 line or the IL-10-expressing line. K-means clustering and functional group analysis revealed a primary up-regulation of ubiquitination enzymes and TNF pathway components and a primary down-regulation of protease inhibitors and protein glycosylation enzymes in CF. Key gene expression changes were confirmed by real-time RT-PCR. Massive reprogramming of gene expression occurred 3 h after Pseudomonas exposure. Changes specific to CF included exaggerated activation of cytokines, blunted activation of anti-proteases, and repression of protein glycosylation enzymes. In conclusion, the CFTR genotype changes the expression of multiple genes at baseline and in response to bacterial challenge, and only a subset of these changes is secondary to IL-10 deficiency.

Comments

Christian Mueller is cited on this publication as Chris Muller. At the time of publication, Christian Mueller and Terence Flotte were not yet affiliated with the University of Massachusetts Medical School.

Citation: Mol Ther. 2004 Sep;10(3):562-73.

Related Resources

Link to Article in PubMed