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 K.

doi:10.1074/jbc.m305788200

Cited by 50 publications

(49 citation statements)

References 38 publications

(54 reference statements)

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“…This may be due to reduced cell death observed during infection with the triple mutant. The cytotoxic enterotoxin (Act) has been reported to cause apoptosis of murine macrophages and induce host inflammatory responses [23,24]. However, in this study, caspase-1 activation and IL-1b release were not affected by mutation of the act gene either alone or in combination with the deletions described above (data not shown).…”

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confidence: 49%

Cytotoxins of the human pathogen Aeromonas hydrophila trigger, via the NLRP3 inflammasome, caspase‐1 activation in macrophages

et al. 2010

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Aeromonas hydrophila is a Gram-negative pathogen that causes serious infectious disease in humans. A. hydrophila induces apoptosis in infected macrophages, but the host proinflammatory responses triggered by macrophage death are largely unknown. Here, we demonstrate that the infection of mouse macrophages with A. hydrophila triggers the activation of caspase-1 and release of IL-1b. Caspase-1 activation was abrogated in macrophages deficient in Nod-like receptor family, pyrin domain containing 3 (NLRP3) and apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), but not NLR family, CARD domain containing 4 (NLRC4). The activation of the NLRP3 inflammasome was mediated by three cytotoxins (aerolysin, hemolysin and multifunctional repeat-in-toxin) produced by A. hydrophila. Our results indicated that the NLRP3 inflammasome senses A. hydrophila infection through the action of bacterial cytotoxins.Key words: Aeromonas . Caspase-1 . Nod-like receptor Supporting Information available online IntroductionThe genus Aeromonas is responsible for a significant number of animal and human infections. Aeromonas are opportunistic human pathogens that can cause intestinal and extraintestinal, wound, soft tissue, skin, and blood infections and septicemia. The most common human disease associated with Aeromonas infection is gastroenteritis with diarrhea symptoms. Among the 21 species that have been differentiated based on DNA-DNA hybridization, A. caviae, A. veronii biotype sobria and A. hydrophila are most commonly associated with human infections and account for more than 85% of all clinical isolates [1,2].Recently, it has been reported that infection with A. hydrophila induces apoptosis in macrophages [3,4]. Although the cytotoxicity by the bacteria is believed to enable the bacteria to evade eradication and clearance by the macrophages, the host proinflammatory responses triggered by the death of the macrophages remain unknown.Induction of apoptosis is considered a virulence mechanism of pathogenic bacteria that can cause tissue damage and Here, we investigated the cell death of macrophages infected with A. hydrophila and demonstrate that the bacteria cause pyroptosis with caspase-1 activation via the NLRP3 inflammasome. Aerolysin, hemolysin (HlyA) and multifunctional repeat-intoxin (RtxA) produced by A. hydrophila are required to elicit the NLRP3 inflammasome and necrotic cell death. We identify the essential bacterial and host factors for eliciting caspase-1 activation and proinflammatory responses. SHORT COMMUNICATION Results and discussionInfection with A. hydrophila induces caspase-1 activation, IL-1b release and pyroptosis It has been reported that aerolysin produced by A. salmonicida induces caspase-1 activation in CHO cells [20,21]. However, whether whole bacterial infection by A. hydrophila leads to caspase-1 activation in macrophages has not been elucidated. We examined caspase-1 activation in infected primary mouse BMderived macrophages (BMM) and observed that BMM infected with WT strai...

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confidence: 49%

Cytotoxins of the human pathogen Aeromonas hydrophila trigger, via the NLRP3 inflammasome, caspase‐1 activation in macrophages

et al. 2010

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“…HT-29 cells were treated with a sublethal dose of Act (12 ng/ml) for 0, 2, and 12 h (in triplicate), VOL. 73, 2005 MICROARRAY AND PROTEOMICS ANALYSES OF HT-29 CELLS 2629 and the RNA was isolated and applied to HU133 GeneChips, as previously described (13). In addition, one experiment (0 and 2 h) of Act-treated (12 ng/ml) polarized HT-29 cells was performed.…”

Section: Resultsmentioning

confidence: 99%

“…Because the host cell receptor for Act could be distributed on either the apical or basolateral surface, polarized cells were treated with the toxin on both sides (apical and basal). The data were analyzed separately using four different techniques: GCOS, SAM, Spotfire 7.3, and ANOVA (13). We expected a fold change of at least 2.0 as significant for Acttreated nonpolarized HT-29 cells (13).…”

Section: Resultsmentioning

confidence: 99%

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Microarray and Proteomics Analyses of Human Intestinal Epithelial Cells Treated with theAeromonas hydrophilaCytotoxic Enterotoxin

et al. 2005

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We performed microarray analyses on RNA from human intestinal epithelial (HT-29) cells treated with the cytotoxic enterotoxin (Act) of Aeromonas hydrophila to examine global cellular transcriptional responses. Based on three independent experiments, Act upregulated the expression of 34 genes involved in cell growth, adhesion, signaling, immune responses (including interleukin-8 [IL-8] production), and apoptosis. We verified the upregulation of 14 genes by real-time reverse transcriptase-PCR and confirmed Act-induced production of IL-8 by enzyme-linked immunosorbent assay on supernatants from nonpolarized and polarized HT-29 cells. Maximal production of IL-8 in response to Act required the presence of intracellular calcium, since chelation of calcium with BAPTA-AM significantly reduced Act-induced IL-8 production in HT-29 cells. We also examined activation of mitogen-activated protein kinases and, as demonstrated by Western blot analysis of apical side-treated polarized HT-29 cells, Act induced phosphorylation of p38, c-Jun NH2-terminal kinase, and extracellular signal-regulated kinase 1/2. In addition, KinetWorks proteomics screening of whole-cell lysates revealed Act-induced phosphorylation of cyclic AMP-response element binding protein (CREB), c-Jun, adducin, protein kinase C, and signal transducer and activator of transcription 3 (STAT3) and decreased phosphorylation of protein kinase Bα, v-raf-1 murine leukemia viral oncogene homolog 1 (i.e., Raf1), and STAT1. We verified activation of CREB and activator protein 1 in polarized cells by gel shift assay. This is the first description of human intestinal epithelial cell transcriptional alterations, phosphorylation or activation of signaling molecules, cytokine production, and calcium mobilization in response to this toxin

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“…Ogawa-Goto et al [39] found that p180, an integral endoplasmic reticulum membrane protein, interacts with a viral protein and that this interaction may play a role in the intracellular transport of the virus. "GALINDO ACT UP" is a set of 88 genes significantly up-regulated by the toxin Act in macrophages [22]. This CDM pattern suggests that the inflammatory response induced by this toxin may include stress to the endoplasmic reticulum.…”

Section: Stress Response In Human Cellsmentioning

confidence: 99%

Compositional mining of multirelational biological datasets

Jin

Murali

Ramakrishnan

2008

ACM Trans. Knowl. Discov. Data

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High-throughput biological screens are yielding ever-growing streams of information about multiple aspects of cellular activity. As more and more categories of datasets come online, there is a corresponding multitude of ways in which inferences can be chained across them, motivating the need for compositional data mining algorithms. In this paper, we argue that such compositional data mining can be effectively realized by functionally cascading redescription mining and biclustering algorithms as primitives. Both these primitives mirror shifts of vocabulary that can be composed in arbitrary ways to create rich chains of inferences. Given a relational database and its schema, we show how the schema can be automatically compiled into a compositional data mining program, and how different domains in the schema can be related through logical sequences of biclustering and redescription invocations. This feature allows us to rapidly prototype new data mining applications, yielding greater understanding of scientific datasets. We describe two applications of compositional data mining: (i) matching terms across categories of the Gene Ontology and (ii) understanding the molecular mechanisms underlying stress response in human cells.

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Identification of Aeromonas hydrophila Cytotoxic Enterotoxin-induced Genes in Macrophages Using Microarrays

Cited by 50 publications

References 38 publications

Cytotoxins of the human pathogen Aeromonas hydrophila trigger, via the NLRP3 inflammasome, caspase‐1 activation in macrophages

Cytotoxins of the human pathogen Aeromonas hydrophila trigger, via the NLRP3 inflammasome, caspase‐1 activation in macrophages

Microarray and Proteomics Analyses of Human Intestinal Epithelial Cells Treated with theAeromonas hydrophilaCytotoxic Enterotoxin

Compositional mining of multirelational biological datasets

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