Effects of CFTR, interleukin-10, and Pseudomonas aeruginosa on gene expression profiles in a CF bronchial epithelial cell Line.
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.
Virella-Lowell
Isabel
I
Department of Pediatrics, Medical University of South Carolina, Charleston, SC 29401, USA.
Herlihy
John-David
JD
Liu
Barry
B
Lopez
Cecilia
C
Cruz
Pedro
P
Muller
Chris
C
Baker
Henry V
HV
Flotte
Terence R
TR
D000818:Animals; D001980:Bronchi / Q000166:cytology / Q000378:metabolism*; D002460:Cell Line; D003550:Cystic Fibrosis / Q000235:genetics; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000096:biosynthesis* / Q000235:genetics; D004847:Epithelial Cells / Q000378:metabolism*; D020869:Gene Expression Profiling; D006801:Humans; D016753:Interleukin-10 / Q000096:biosynthesis* / Q000235:genetics; D009154:Mutation; D011550:Pseudomonas aeruginosa; D020545:Respiratory Mucosa / Q000166:cytology / Q000378:metabolism; D014162:Transfection
D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D016753:Interleukin-10