Identification of Aeromonas hydrophila cytotoxic enterotoxin-induced genes in macrophages using microarrays

Cristi L. Galindo, Jian Sha, Deborah A. Ribardo, Amin A. Fadl, Lakshmi Pillai, Ashok Chopra

Research output: Contribution to journalArticle

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Abstract

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 h from Act-treated macrophages and applied to Affymetrix MGU74 arrays, and the data were processed using a multi-analysis approach to identify genes that might be critical in the inflammatory process evoked by Act. Seventy-six genes were significantly and consistently upregulated. 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 upregulated 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 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.

Original languageEnglish (US)
Pages (from-to)40198-40212
Number of pages15
JournalJournal of Biological Chemistry
Volume278
Issue number41
DOIs
StatePublished - Oct 10 2003
Externally publishedYes

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Aeromonas hydrophila
Macrophages
Enterotoxins
Microarrays
Genes
Apoptosis
Polymerase chain reaction
RNA-Directed DNA Polymerase
Reverse Transcriptase Polymerase Chain Reaction
Signal transduction
Immunosorbents
Annexin A5
Bioactivity
Gene expression
Microarray Analysis
Assays
Transcription Factors
Adhesion
Real-Time Polymerase Chain Reaction
Signal Transduction

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification of Aeromonas hydrophila cytotoxic enterotoxin-induced genes in macrophages using microarrays. / Galindo, Cristi L.; Sha, Jian; Ribardo, Deborah A.; Fadl, Amin A.; Pillai, Lakshmi; Chopra, Ashok.

In: Journal of Biological Chemistry, Vol. 278, No. 41, 10.10.2003, p. 40198-40212.

Research output: Contribution to journalArticle

Galindo, Cristi L. ; Sha, Jian ; Ribardo, Deborah A. ; Fadl, Amin A. ; Pillai, Lakshmi ; Chopra, Ashok. / Identification of Aeromonas hydrophila cytotoxic enterotoxin-induced genes in macrophages using microarrays. In: Journal of Biological Chemistry. 2003 ; Vol. 278, No. 41. pp. 40198-40212.
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