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Nucleotide Binding Oligomerization Domain 1 Is an Essential Signal Transducer in Human Epithelial Cells Infected with Helicobacter pylori That Induces the Transepithelial Migration of Neutrophils.

Kim BJ, Kim JY, Hwang ES, Kim JG - Gut Liver (2015)

Bottom Line: Cytotoxin-associated gene pathogenicity island (cagPAI)(+) H. pylori infection upregulated IL-8 mRNA expression and IL-8 secretion in AGS and Caco-2 cells compared with controls.NF-κB activation, IL-8 mRNA expression and IL-8 secretion by cagPAI knockdown strains were reduced compared with those infected with the wild-type strain.NF-κB activation, IL-8 mRNA expression and IL-8 secretion in dominant-negative (DN)-Nod1 stably transfected cells were reduced compared with the controls.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine, Chung-Ang University College of Medicine, Seoul, Korea.

ABSTRACT

Background/aims: The cytosolic host protein nucleotide binding oligomerization domain 1 (Nod1) has emerged as a key pathogen recognition molecule for innate immune responses in epithelial cells. The purpose of the study was to elucidate the mechanism by which Helicobacter pylori infection leads to transepithelial neutrophil migration in a Nod1-mediated manner.

Methods: Human epithelial cell lines AGS and Caco-2 were grown and infected with H. pylori. Interleukin (IL)-8 mRNA expression and IL-8 secretion were assessed, and nuclear factor κB (NF-κB) activation was determined. Stable transfections of AGS and Caco-2 cells with dominant negative Nod1 were generated. Neutrophil migration across the monolayer was quantified.

Results: Cytotoxin-associated gene pathogenicity island (cagPAI)(+) H. pylori infection upregulated IL-8 mRNA expression and IL-8 secretion in AGS and Caco-2 cells compared with controls. NF-κB activation, IL-8 mRNA expression and IL-8 secretion by cagPAI knockdown strains were reduced compared with those infected with the wild-type strain. NF-κB activation, IL-8 mRNA expression and IL-8 secretion in dominant-negative (DN)-Nod1 stably transfected cells were reduced compared with the controls. The transepithelial migration of neutrophils in DN-Nod1 stably transfected cells was reduced compared with that in controls.

Conclusions: Signaling through Nod1 plays an essential role in neutrophil migration induced by the upregulated NF-κB activation and IL-8 expression in H. pylori-infected human epithelial cells.

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Interleukin (IL)-8 mRNA and protein expression in dominant-negative nucleotide binding oligomerization domain 1 (DN-Nod1) stably transfected AGS cells infected with cytotoxin-associated gene pathogenicity island (cagPAI)-depleted Helicobacter pylori. (A) Decreased expression of IL-8 mRNA in DN-Nod1 stably transfected AGS cells infected with the cagPAI knockdown strain was observed compared with the expression levels in cells infected with wild-type ATCC 60190. (B) IL-8 mRNA expression in DN-Nod1 stably transfected AGS cells infected with cagPAI-depleted H. pylori was reduced compared with the expression in cells infected with wild-type ATCC 60190, as determined by real-time reverse transcription-polymerase chain reaction analysis. (C) DN-Nod1 stably transfected AGS cells infected with the cagPAI knockdown strain displayed a reduced capacity to mediate IL-8 secretion compared with cells infected with wild-type ATCC 60190. The error bars indicate the standard error of the mean of triplicate samples, which were representative of three independent experiments. *p<0.05.
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f7-gnl-09-358: Interleukin (IL)-8 mRNA and protein expression in dominant-negative nucleotide binding oligomerization domain 1 (DN-Nod1) stably transfected AGS cells infected with cytotoxin-associated gene pathogenicity island (cagPAI)-depleted Helicobacter pylori. (A) Decreased expression of IL-8 mRNA in DN-Nod1 stably transfected AGS cells infected with the cagPAI knockdown strain was observed compared with the expression levels in cells infected with wild-type ATCC 60190. (B) IL-8 mRNA expression in DN-Nod1 stably transfected AGS cells infected with cagPAI-depleted H. pylori was reduced compared with the expression in cells infected with wild-type ATCC 60190, as determined by real-time reverse transcription-polymerase chain reaction analysis. (C) DN-Nod1 stably transfected AGS cells infected with the cagPAI knockdown strain displayed a reduced capacity to mediate IL-8 secretion compared with cells infected with wild-type ATCC 60190. The error bars indicate the standard error of the mean of triplicate samples, which were representative of three independent experiments. *p<0.05.

Mentions: As shown in Fig. 7B, decreased expression of IL-8 mRNA in DN-Nod1 stably trasfected AGS cells infected with cagPAI knockdown strain was observed in real-time RT-PCR assay. In addition, ELISA showed that DN-Nod1 stably trasfected AGS cells infected with cagPAI knockdown strain displayed a reduced ability to mediate IL-8 release, compared with those infected with wild type ATCC 60190 (Fig. 7C).


Nucleotide Binding Oligomerization Domain 1 Is an Essential Signal Transducer in Human Epithelial Cells Infected with Helicobacter pylori That Induces the Transepithelial Migration of Neutrophils.

Kim BJ, Kim JY, Hwang ES, Kim JG - Gut Liver (2015)

Interleukin (IL)-8 mRNA and protein expression in dominant-negative nucleotide binding oligomerization domain 1 (DN-Nod1) stably transfected AGS cells infected with cytotoxin-associated gene pathogenicity island (cagPAI)-depleted Helicobacter pylori. (A) Decreased expression of IL-8 mRNA in DN-Nod1 stably transfected AGS cells infected with the cagPAI knockdown strain was observed compared with the expression levels in cells infected with wild-type ATCC 60190. (B) IL-8 mRNA expression in DN-Nod1 stably transfected AGS cells infected with cagPAI-depleted H. pylori was reduced compared with the expression in cells infected with wild-type ATCC 60190, as determined by real-time reverse transcription-polymerase chain reaction analysis. (C) DN-Nod1 stably transfected AGS cells infected with the cagPAI knockdown strain displayed a reduced capacity to mediate IL-8 secretion compared with cells infected with wild-type ATCC 60190. The error bars indicate the standard error of the mean of triplicate samples, which were representative of three independent experiments. *p<0.05.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4413970&req=5

f7-gnl-09-358: Interleukin (IL)-8 mRNA and protein expression in dominant-negative nucleotide binding oligomerization domain 1 (DN-Nod1) stably transfected AGS cells infected with cytotoxin-associated gene pathogenicity island (cagPAI)-depleted Helicobacter pylori. (A) Decreased expression of IL-8 mRNA in DN-Nod1 stably transfected AGS cells infected with the cagPAI knockdown strain was observed compared with the expression levels in cells infected with wild-type ATCC 60190. (B) IL-8 mRNA expression in DN-Nod1 stably transfected AGS cells infected with cagPAI-depleted H. pylori was reduced compared with the expression in cells infected with wild-type ATCC 60190, as determined by real-time reverse transcription-polymerase chain reaction analysis. (C) DN-Nod1 stably transfected AGS cells infected with the cagPAI knockdown strain displayed a reduced capacity to mediate IL-8 secretion compared with cells infected with wild-type ATCC 60190. The error bars indicate the standard error of the mean of triplicate samples, which were representative of three independent experiments. *p<0.05.
Mentions: As shown in Fig. 7B, decreased expression of IL-8 mRNA in DN-Nod1 stably trasfected AGS cells infected with cagPAI knockdown strain was observed in real-time RT-PCR assay. In addition, ELISA showed that DN-Nod1 stably trasfected AGS cells infected with cagPAI knockdown strain displayed a reduced ability to mediate IL-8 release, compared with those infected with wild type ATCC 60190 (Fig. 7C).

Bottom Line: Cytotoxin-associated gene pathogenicity island (cagPAI)(+) H. pylori infection upregulated IL-8 mRNA expression and IL-8 secretion in AGS and Caco-2 cells compared with controls.NF-κB activation, IL-8 mRNA expression and IL-8 secretion by cagPAI knockdown strains were reduced compared with those infected with the wild-type strain.NF-κB activation, IL-8 mRNA expression and IL-8 secretion in dominant-negative (DN)-Nod1 stably transfected cells were reduced compared with the controls.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine, Chung-Ang University College of Medicine, Seoul, Korea.

ABSTRACT

Background/aims: The cytosolic host protein nucleotide binding oligomerization domain 1 (Nod1) has emerged as a key pathogen recognition molecule for innate immune responses in epithelial cells. The purpose of the study was to elucidate the mechanism by which Helicobacter pylori infection leads to transepithelial neutrophil migration in a Nod1-mediated manner.

Methods: Human epithelial cell lines AGS and Caco-2 were grown and infected with H. pylori. Interleukin (IL)-8 mRNA expression and IL-8 secretion were assessed, and nuclear factor κB (NF-κB) activation was determined. Stable transfections of AGS and Caco-2 cells with dominant negative Nod1 were generated. Neutrophil migration across the monolayer was quantified.

Results: Cytotoxin-associated gene pathogenicity island (cagPAI)(+) H. pylori infection upregulated IL-8 mRNA expression and IL-8 secretion in AGS and Caco-2 cells compared with controls. NF-κB activation, IL-8 mRNA expression and IL-8 secretion by cagPAI knockdown strains were reduced compared with those infected with the wild-type strain. NF-κB activation, IL-8 mRNA expression and IL-8 secretion in dominant-negative (DN)-Nod1 stably transfected cells were reduced compared with the controls. The transepithelial migration of neutrophils in DN-Nod1 stably transfected cells was reduced compared with that in controls.

Conclusions: Signaling through Nod1 plays an essential role in neutrophil migration induced by the upregulated NF-κB activation and IL-8 expression in H. pylori-infected human epithelial cells.

Show MeSH
Related in: MedlinePlus