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Papers In Press, published online ahead of print June 24, 2003
Microbiology & Immunology, University of Texas Medical Branch at Galveston, Galveston, Texas 77555
Corresponding Author: clgalind{at}utmb.edu
A cytotoxic enterotoxin (Act) of Aeromonas hydrophila possesses several biological activities, and it induces an inflammatory response in the host. In this study, we used microarrays to gain a global and molecular view of the cellular transcriptional responses to Act and to identify important genes up-regulated by this toxin. Total RNA was isolated at 0, 2, and 12 hr from Act-treated macrophages, was applied to Affymetrix MGU74 arrays, and data processed by using a multi-analysis approach in order to identify genes that might be critical in the inflammatory process evoked by Act. Seventy-eight genes were significantly and consistently up-regulated. Many of these genes were immune-related, and several were transcription factors, adhesion molecules, and cytokines. Additionally, we identified several apoptosis-associated genes that were significantly up-regulated in Act-treated macrophages. Act-induced apoptosis of macrophages was confirmed by annexin V staining and DNA laddering. Quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) were used to verify increased expression of some inflammatory and apoptosis-associated genes identified by the microarray analysis. To further confirm Act-induced increases in gene expression, real-time RT-PCR was also used for selected genes. Taken together, the array data provided for the first time a global view of Act-mediated signal transduction and clearly demonstrated an inflammatory response and apoptosis mediated by this toxin in host cells at the molecular level.
J. Biol. Chem, 10.1074/jbc.M305788200
Submitted on June 3, 2003
Revised on June 20, 2003
Accepted on June 24, 2003
Identification of Aeromonas hydrophila cytotoxic enterotoxin-induced genes in macrophages using microarrays
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