Activation of NF-κB via the IκB Kinase Complex Is Both Essential and Sufficient for Proinflammatory Gene Expression in Primary Endothelial Cells

Denk, Andrea; Goebeler, Matthias; Schmid, Sybille; Berberich, Ingolf; Ritz, Olga; Lindemann, Dirk; Ludwig, Stephan; Wirth, Thomas

doi:10.1074/jbc.m102698200

Cited by 191 publications

(179 citation statements)

References 41 publications

Supporting

Mentioning

169

Contrasting

Unclassified

Order By: Relevance

“…The pCFG5-IEGZ retroviral vector containing an insert of kinase-dead IKK2 (IKK2KD) coupled to expression of a GFP-zeocin resistance fusion gene through an internal ribosomal entry site has been described earlier (11). A dominant negative mutant of human MyD88 (aa 152-296) was provided by Tularik Inc. (South San Francisco, CA) and cloned into the pCFG5-IEGZ retroviral vector.…”

Section: Plasmidsmentioning

confidence: 99%

Candida albicans Triggers Activation of Distinct Signaling Pathways to Establish a Proinflammatory Gene Expression Program in Primary Human Endothelial Cells

Müller

Viemann²,

Schmidt

et al. 2007

The Journal of Immunology

Self Cite

View full text Add to dashboard Cite

Endothelial cells (EC) actively participate in the innate defense against microbial pathogens. Under unfavorable conditions, defense reactions can turn life threatening resulting in sepsis. We therefore studied the so far largely unknown EC reaction patterns to the fungal pathogen Candida albicans, which is a major cause of lethality in septic patients. Using oligonucleotide microarray analysis, we identified 56 genes that were transcriptionally up-regulated and 69 genes that were suppressed upon exposure of ECs to C. albicans. The most significantly up-regulated transcripts were found in gene ontology groups comprising the following categories: chemotaxis/migration; cell death and proliferation; signaling; transcriptional regulation; and cell-cell contacts/intercellular signaling. Further examination of candidate signaling cascades established a central role of the proinflammatory NF-κB pathway in the regulation of the Candida-modulated transcriptome of ECs. As a second major regulatory pathway we identified the stress-activated p38 MAPK pathway, which critically contributes to the regulation of selected Candida target genes such as CXCL8/IL-8. Depletion of MyD88 and IL-1R-associated kinase-1 by RNA interference demonstrates that Candida-induced NF-κB activation is mediated by pattern recognition receptor signaling. Additional experiments suggest that C. albicans-induced CXCL8/IL-8 expression is mediated by TLR3 rather than TLR2 and TLR4, which previously have been implicated with MyD88/IκB kinase-2/NF-κB activation by this fungus in other systems. Our study provides the first comprehensive analysis of endothelial gene responses to C. albicans and presents novel insights into the complex signaling patterns triggered by this important pathogen.

show abstract

Section: Plasmidsmentioning

confidence: 99%

Candida albicans Triggers Activation of Distinct Signaling Pathways to Establish a Proinflammatory Gene Expression Program in Primary Human Endothelial Cells

Müller

Viemann²,

Schmidt

et al. 2007

The Journal of Immunology

Self Cite

View full text Add to dashboard Cite

show abstract

“…The pCFG5-IEGZ retroviral vector system, as described by Denk et al (2001), was used to infect U87MG and T98G glioblastoma cells. PT67 producer cells (Clontech, Palo Alto, CA, USA) were transfected with empty pCFG5-IEGZ vectors or pCFG5-IEGZ vectors containing IkB-a-(S32; 36A) using Lipofectamine 2000 (Invitrogen) according to the manufacturer's recommendation.…”

Section: Production Of Retrovirus and Retroviral Transductionmentioning

confidence: 99%

NF-κB-independent sensitization of glioblastoma cells for TRAIL-induced apoptosis by proteasome inhibition

et al. 2006

View full text Add to dashboard Cite

“…1,39 Transcriptional activation of the genes encoding these adhesion molecules and cytokines is tightly regulated by a transcription factor, NFkB. 4,6,[9][10][11]13,40,41 Under normal conditions, NFkB is usually present with IkB in the cytosol as an inactive complex. However, when grafts are exposed to stimuli after transplantation such as ischemia/reperfusion injury, cells may be activated to promote IkB-phosphorylation, which induces NFkB to translocate to the nucleus and to bind to the promoter region of genes of several major inflammatory mediators, such as IL-1, iNOS, and TNF-a, and upregulate their transcription.…”

Section: Discussionmentioning

confidence: 99%

“…However, when grafts are exposed to stimuli after transplantation such as ischemia/reperfusion injury, cells may be activated to promote IkB-phosphorylation, which induces NFkB to translocate to the nucleus and to bind to the promoter region of genes of several major inflammatory mediators, such as IL-1, iNOS, and TNF-a, and upregulate their transcription. [5][6][7][8][9][10][11][12][13][14] These products may also promote NFkB activation in a paracrine or autocrine fashion, and this in turn further upregulates the production of chemokines and the expression of surface molecules, establishing the so-called 'cytokine-adhesion molecule cascade', which further augments such inflammatory reaction and promotes the development of acute rejection. [41][42][43][44][45] As we previously reported, NFkB-decoy, which effectively binds NFkB and inhibits its activity, blocks the transactivation of genes for essential cytokines and adhesion molecules in vivo in several animal models of inflammatory disease as well as in vitro.…”

Section: Discussionmentioning

confidence: 99%

“…2,3 Nuclear factor kB (NFkB) is one of such transcription factors for genes whose products are critical for inflammation and immunity. 4 It rapidly translocates from the cytoplasm to the nucleus in response to a variety of extracellular signals and plays a pivotal role in the regulation of inflammatory changes by inducing coordinated transactivation of genes of major inflammatory mediators such as IL-1, IL-6, iNOS, and TNF-a [5][6][7][8][9][10][11] and adhesion molecules such as ICAM-1 and VCAM-1. [12][13][14] Cooper et al demonstrated a time-dependent increase in the DNA-binding activity of NFkB, which peaked at 3 days before rejection, in a rat cardiac allograft model.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Transfection of NFκB-decoy oligodeoxynucleotides using efficient ultrasound-mediated gene transfer into donor kidneys prolonged survival of rat renal allografts

et al. 2003

View full text Add to dashboard Cite

Nuclear factor kB (NFkB) plays a pivotal role in the coordinated transactivation of a series of genes of cytokines and adhesion molecules that are highly involved in the onset of acute rejection in organ transplantation. We previously developed decoy cis-elements oligo deoxyribonucleic acid against NFkB (NFkB-decoy) that effectively inhibited the activation of major inflammatory mediators in vitro and in vivo. Accordingly, we hypothesized that transfection of NFkB-decoy into the donor kidney would prevent acute rejection and prolong graft survival, and thus provide effective therapy for renal acute rejection. To transfect NFkB-decoy, we employed a novel approach using ultrasound exposure with an echocardiographic contrast agent, Optison, and clearly demonstrated successful transfection of NFkB-decoy into renal tissue. The therapeutic effect of NFkB-decoy on renal allografts was then evaluated in a rat renal allograft model (Wistar-Lewis). In the control group, graft function significantly deteriorated with marked destruction of renal tissue, accompanied by increased production of major inflammatory mediators, and all animals died of renal failure by 9 days. In contrast, graft function (serum creatinine on day 2, NFkB-treated: 0.9770.16 versus control: 1.8470.23 mg/dl, Po0.01) and histological structure were well preserved with significantly decreased expression of NFkB-regulated cytokines and adhesion molecules, including IL-1, iNOS, MCP-1, TNF-a, and ICAM-1, in allografts transfected with NFkB-decoy. As a result, animal survival was significantly prolonged in this group as compared to controls (14.275.2 versus 7.171.2 days, Po0.01). Thus, we established a novel ultrasound-Optison-mediated gene transfection approach and demonstrated the significant prolongation of graft survival by the successful transfection of NFkB-decoy into the donor kidney in a rat renal allograft model. Gene Therapy (2003) 10, 415-425.

show abstract

Activation of NF-κB via the IκB Kinase Complex Is Both Essential and Sufficient for Proinflammatory Gene Expression in Primary Endothelial Cells

Cited by 191 publications

References 41 publications

Candida albicans Triggers Activation of Distinct Signaling Pathways to Establish a Proinflammatory Gene Expression Program in Primary Human Endothelial Cells

Candida albicans Triggers Activation of Distinct Signaling Pathways to Establish a Proinflammatory Gene Expression Program in Primary Human Endothelial Cells

NF-κB-independent sensitization of glioblastoma cells for TRAIL-induced apoptosis by proteasome inhibition

Transfection of NFκB-decoy oligodeoxynucleotides using efficient ultrasound-mediated gene transfer into donor kidneys prolonged survival of rat renal allografts

Contact Info

Product

Resources

About