Fig 1.
Validation of activation of primary bronchial epithelial cells (hAEBCs) upon P. aeruginosa infection.
Kinetics of IL-8 (A) and IL-6 (B) production by hAEBCs infected with P. aeruginosa. At each time point, supernatants were collected for ELISAs of IL-8 and IL-6. The data are mean±SEM of IL-8 or IL-6 concentrations for four different patients. The statistical analysis consisted in ANOVA followed by Bonferroni’s multiple comparison test. *P<0.05 and **P<0.001. Each value in the cystic-fibrosis (CF) group was compared with the value at the same time point in the CTRL group.
Table 1.
List of genes in the protein-binding gene-ontology category that were most upregulated (FC ≥ 3) in CTRL cells 6 hours after P. aeruginosa infection.
Fig 2.
Overview of gene expression profiles.
A- The heat-map of the mean expression levels of all the genes in each condition revealed two distinct clusters that separated cystic-fibrosis (CF) cells from CTRL cells. B and C- Venn diagram of differentially expressed genes between cystic-fibrosis and CTRL cells upon P. aeruginosa infection. Circles: Differentially expressed genes upregulated (B) or downregulated (C) at a single postinfection time point or at two or three consecutive postinfection time points (thus, 0h was not considered). We selected the genes whose fold-change in expression level was ≥2 in the event of upregulation and ≤0.5 in the event of downregulation. Squares: Differentially expressed genes upregulated (B) or downregulated (C) at two, three, or four consecutive time points (thus, 0h was considered). We selected the genes for which the ratio of FC in infected cells over FC in noninfected cells was ≥1.5 in the event of upregulation or ≤0.6 in the event of downregulation.
Table 2.
Gene-ontology molecular-function categories upregulated in infected cystic-fibrosis cells compared to CTRL cells.
Table 3.
Genes in the protein-binding (A) and in the catalytic-activity (B) gene-ontology category: five upregulated with the greatest difference in expression level between cystic-fibrosis and CTRL cells after infection.
Fig 3.
TNF (A), CSF3 (B), CSF2 (C), CCL2 (D), MMP1 (E), and MMP10 (F) protein synthesis in response to P. aeruginosa infection of cystic-fibrosis (CF) and CTRL cells.
Protein levels were measured in supernatants of infected CF and CTRL cells 0, 2, 4, 6, and 8 hours postinfection, using the Human Magnetic Luminex Screening Assay. Data are mean±SEM of cytokine concentrations for four different patients. The statistical analysis consisted in ANOVA followed by Bonferroni’s multiple comparison test. *P<0.05 and **P<0.001. Each value in the cystic-fibrosis (CF) group was compared with the value at the same time point in the CTRL group.
Table 4.
Gene-ontology molecular-function categories downregulated in infected cystic-fibrosis cells compared to CTRL cells.
Table 5.
Genes in the catalytic-activity (A) and in the protein-binding (B) gene-ontology category: five downregulated genes with the greatest difference in expression level between cystic-fibrosis and CTRL cells after infection.