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Physiol. Genomics (December 14, 2004). doi:10.1152/physiolgenomics.00135.2004 Free Article
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Submitted on June 10, 2004
Accepted on December 9, 2004

Live Staphylococcus aureus and Bacterial Soluble Factors Induce Different Transcriptional Responses in Human Airway Cells

Chimene Moreilhon1, Delphine Gras2, Coralie Hologne2, Odile Bajolet2, Francoise Cottrez3, Virginie Magnone1, Marc Merten4, Herve Groux3, Edith Puchelle2, and Pascal Barbry1*

1 UMR 6097, CNRS, Institut de Pharmacologie Moleculaire et Cellulaire, Sophia-Antipolis, France
2 INSERM UMR-S 514, CHU Hopital Maison Blanche, Reims, France
3 INSERM U576, Hopital de l'Archet, Nice, France
4 INSERM EMI 10014, Vandoeuvre-les-Nancy, France

* To whom correspondence should be addressed. E-mail: barbry{at}ipmc.cnrs.fr.

To characterize the response of respiratory epithelium to infection by Staphylococcus aureus (S. aureus), human airway cells were incubated for 1 to 24 hours with a supernatant of a S. aureus culture (bacterial supernatant), then profiled with a pangenomic DNA microarray. Because an up-regulation of many genes was noticed around three hours, three independent approaches were then used to characterize the host response to a three-hour contact either with bacterial supernatant or with live bacteria: (i) a DNA microarray containing 4200 sequence-verified probes, (ii) a semi quantitative RT-PCR with a set of 537 pairs of validated primers, (iii) ELISA assay of IL-8, IL-6, TNF alpha and PGE2. Among others, Fos, Jun and Egr-1 were up-regulated by the bacterial supernatant and by live bacteria. Increased expression of bhlhb2 and Mig-6, which promoter regions harbour HIF responding elements, was explained by an increased expression of the HIF-1{alpha} protein. Activation of the inducible form of cyclo-oxygenase COX-2 and of the interleukins IL-1, IL-6 and IL-8 as well as of the NF-{kappa}B pathway was observed preferentially in cells in contact with bacterial supernatant. Early infection was characterized by an up-regulation of anti-apoptotic genes and a down-regulation of pro-apoptotic genes. This correlated with a necrotic, rather than apoptotic cell death. Overall, this first global description of an airway epithelial infection by S. aureus demonstrates a larger global response to bacterial supernatant (in term of altered genes and variation factors) than to exponentially growing live bacteria.




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