Molecular characterization of Helicobacter pylori vacA induction of IL-8 in U937 cells reveals a prominent role for p38MAPK in activating transcription factor-2, cAMP response element binding protein, and NF-κB Activation

Junzo Hisatsune, Masaaki Nakayama, Hajime Isomoto, Hisao Kurazono, Naofumi Mukaida, Asish K. Mukhopadhyay, Takeshi Azuma, Yoshio Yamaoka, Jan Sap, Eiki Yamasaki, Kinnosuke Yahiro, Joel Moss, Toshiya Hirayama

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Abstract

Helicobacter pylori VacA induces multiple effects on susceptible cells, including vacuolation, mitochondrial damage, inhibition of cell growth, and enhanced cyclooxygenase-2 expression. To assess the ability of H. pylori to modulate the production of inflammatory mediators, we examined the mechanisms by which VacA enhanced IL-8 production by promonocytic U937 cells, which demonstrated the greatest VacA-induced IL-8 release of the cells tested. Inhibitors of p38 MAPK (SB203580), ERK1/2 (PD98059), IkBα ((E)-3-(4-methylphenylsulfonyl)-2-propenenitrile), Ca 2+ entry (SKF96365), and intracellular Ca 2+ channels (dantrolene) blocked VacA-induced IL-8 production. Furthermore, an intracellular Ca 2+ chelator (BAPTA-AM), which inhibited VacA-acti- vated p38 MAPK, caused a dose-dependent reduction in VacA-induced IL-8 secretion by U937 cells, implying a role for intra- cellular Ca 2+ in mediating activation of MAPK and the canonical NF-κB pathway. VacA stimulated translocation of NF-κBp65 to the nucleus, consistent with enhancement of IL-8 expression by activation of the NF-κB pathway. In addition, small interfering RNA of activating transcription factor (ATF)-2 or CREB, which is a p38MAPK substrate and binds to the AP-1 site of the IL-8 promoter, inhibited VacA-induced IL-8 production. VacA activated an IL-8 promoter containing an NF-IL-6 site, but not a mutated AP-1 or NF-κB site, suggesting direct involvement of the ATF-2/CREB binding region or NF-κB-binding regions in VacA-induced IL-8 promoter activation. Thus, in U937 cells, VacA directly increases IL-8 production by activation of the p38 MAPK via intracellular Ca 2+ release, leading to activation of the transcription factors, ATF-2, CREB, and NF-κB.

Original languageEnglish
Pages (from-to)5017-5027
Number of pages11
JournalJournal of Immunology
Volume180
Issue number7
Publication statusPublished - Apr 1 2008
Externally publishedYes

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Activating Transcription Factor 2
Cyclic AMP Response Element-Binding Protein
U937 Cells
Interleukin-8
Helicobacter pylori
p38 Mitogen-Activated Protein Kinases
1-(2-(3-(4-methoxyphenyl)propoxy)-4-methoxyphenylethyl)-1H-imidazole
Transcription Factor AP-1
IgA receptor
Dantrolene
Cyclooxygenase 2
Chelating Agents
Small Interfering RNA
Interleukin-6
Transcription Factors

ASJC Scopus subject areas

  • Immunology

Cite this

Molecular characterization of Helicobacter pylori vacA induction of IL-8 in U937 cells reveals a prominent role for p38MAPK in activating transcription factor-2, cAMP response element binding protein, and NF-κB Activation. / Hisatsune, Junzo; Nakayama, Masaaki; Isomoto, Hajime; Kurazono, Hisao; Mukaida, Naofumi; Mukhopadhyay, Asish K.; Azuma, Takeshi; Yamaoka, Yoshio; Sap, Jan; Yamasaki, Eiki; Yahiro, Kinnosuke; Moss, Joel; Hirayama, Toshiya.

In: Journal of Immunology, Vol. 180, No. 7, 01.04.2008, p. 5017-5027.

Research output: Contribution to journalArticle

Hisatsune, J, Nakayama, M, Isomoto, H, Kurazono, H, Mukaida, N, Mukhopadhyay, AK, Azuma, T, Yamaoka, Y, Sap, J, Yamasaki, E, Yahiro, K, Moss, J & Hirayama, T 2008, 'Molecular characterization of Helicobacter pylori vacA induction of IL-8 in U937 cells reveals a prominent role for p38MAPK in activating transcription factor-2, cAMP response element binding protein, and NF-κB Activation', Journal of Immunology, vol. 180, no. 7, pp. 5017-5027.
Hisatsune, Junzo ; Nakayama, Masaaki ; Isomoto, Hajime ; Kurazono, Hisao ; Mukaida, Naofumi ; Mukhopadhyay, Asish K. ; Azuma, Takeshi ; Yamaoka, Yoshio ; Sap, Jan ; Yamasaki, Eiki ; Yahiro, Kinnosuke ; Moss, Joel ; Hirayama, Toshiya. / Molecular characterization of Helicobacter pylori vacA induction of IL-8 in U937 cells reveals a prominent role for p38MAPK in activating transcription factor-2, cAMP response element binding protein, and NF-κB Activation. In: Journal of Immunology. 2008 ; Vol. 180, No. 7. pp. 5017-5027.
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abstract = "Helicobacter pylori VacA induces multiple effects on susceptible cells, including vacuolation, mitochondrial damage, inhibition of cell growth, and enhanced cyclooxygenase-2 expression. To assess the ability of H. pylori to modulate the production of inflammatory mediators, we examined the mechanisms by which VacA enhanced IL-8 production by promonocytic U937 cells, which demonstrated the greatest VacA-induced IL-8 release of the cells tested. Inhibitors of p38 MAPK (SB203580), ERK1/2 (PD98059), IkBα ((E)-3-(4-methylphenylsulfonyl)-2-propenenitrile), Ca 2+ entry (SKF96365), and intracellular Ca 2+ channels (dantrolene) blocked VacA-induced IL-8 production. Furthermore, an intracellular Ca 2+ chelator (BAPTA-AM), which inhibited VacA-acti- vated p38 MAPK, caused a dose-dependent reduction in VacA-induced IL-8 secretion by U937 cells, implying a role for intra- cellular Ca 2+ in mediating activation of MAPK and the canonical NF-κB pathway. VacA stimulated translocation of NF-κBp65 to the nucleus, consistent with enhancement of IL-8 expression by activation of the NF-κB pathway. In addition, small interfering RNA of activating transcription factor (ATF)-2 or CREB, which is a p38MAPK substrate and binds to the AP-1 site of the IL-8 promoter, inhibited VacA-induced IL-8 production. VacA activated an IL-8 promoter containing an NF-IL-6 site, but not a mutated AP-1 or NF-κB site, suggesting direct involvement of the ATF-2/CREB binding region or NF-κB-binding regions in VacA-induced IL-8 promoter activation. Thus, in U937 cells, VacA directly increases IL-8 production by activation of the p38 MAPK via intracellular Ca 2+ release, leading to activation of the transcription factors, ATF-2, CREB, and NF-κB.",
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AU - Hisatsune, Junzo

AU - Nakayama, Masaaki

AU - Isomoto, Hajime

AU - Kurazono, Hisao

AU - Mukaida, Naofumi

AU - Mukhopadhyay, Asish K.

AU - Azuma, Takeshi

AU - Yamaoka, Yoshio

AU - Sap, Jan

AU - Yamasaki, Eiki

AU - Yahiro, Kinnosuke

AU - Moss, Joel

AU - Hirayama, Toshiya

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N2 - Helicobacter pylori VacA induces multiple effects on susceptible cells, including vacuolation, mitochondrial damage, inhibition of cell growth, and enhanced cyclooxygenase-2 expression. To assess the ability of H. pylori to modulate the production of inflammatory mediators, we examined the mechanisms by which VacA enhanced IL-8 production by promonocytic U937 cells, which demonstrated the greatest VacA-induced IL-8 release of the cells tested. Inhibitors of p38 MAPK (SB203580), ERK1/2 (PD98059), IkBα ((E)-3-(4-methylphenylsulfonyl)-2-propenenitrile), Ca 2+ entry (SKF96365), and intracellular Ca 2+ channels (dantrolene) blocked VacA-induced IL-8 production. Furthermore, an intracellular Ca 2+ chelator (BAPTA-AM), which inhibited VacA-acti- vated p38 MAPK, caused a dose-dependent reduction in VacA-induced IL-8 secretion by U937 cells, implying a role for intra- cellular Ca 2+ in mediating activation of MAPK and the canonical NF-κB pathway. VacA stimulated translocation of NF-κBp65 to the nucleus, consistent with enhancement of IL-8 expression by activation of the NF-κB pathway. In addition, small interfering RNA of activating transcription factor (ATF)-2 or CREB, which is a p38MAPK substrate and binds to the AP-1 site of the IL-8 promoter, inhibited VacA-induced IL-8 production. VacA activated an IL-8 promoter containing an NF-IL-6 site, but not a mutated AP-1 or NF-κB site, suggesting direct involvement of the ATF-2/CREB binding region or NF-κB-binding regions in VacA-induced IL-8 promoter activation. Thus, in U937 cells, VacA directly increases IL-8 production by activation of the p38 MAPK via intracellular Ca 2+ release, leading to activation of the transcription factors, ATF-2, CREB, and NF-κB.

AB - Helicobacter pylori VacA induces multiple effects on susceptible cells, including vacuolation, mitochondrial damage, inhibition of cell growth, and enhanced cyclooxygenase-2 expression. To assess the ability of H. pylori to modulate the production of inflammatory mediators, we examined the mechanisms by which VacA enhanced IL-8 production by promonocytic U937 cells, which demonstrated the greatest VacA-induced IL-8 release of the cells tested. Inhibitors of p38 MAPK (SB203580), ERK1/2 (PD98059), IkBα ((E)-3-(4-methylphenylsulfonyl)-2-propenenitrile), Ca 2+ entry (SKF96365), and intracellular Ca 2+ channels (dantrolene) blocked VacA-induced IL-8 production. Furthermore, an intracellular Ca 2+ chelator (BAPTA-AM), which inhibited VacA-acti- vated p38 MAPK, caused a dose-dependent reduction in VacA-induced IL-8 secretion by U937 cells, implying a role for intra- cellular Ca 2+ in mediating activation of MAPK and the canonical NF-κB pathway. VacA stimulated translocation of NF-κBp65 to the nucleus, consistent with enhancement of IL-8 expression by activation of the NF-κB pathway. In addition, small interfering RNA of activating transcription factor (ATF)-2 or CREB, which is a p38MAPK substrate and binds to the AP-1 site of the IL-8 promoter, inhibited VacA-induced IL-8 production. VacA activated an IL-8 promoter containing an NF-IL-6 site, but not a mutated AP-1 or NF-κB site, suggesting direct involvement of the ATF-2/CREB binding region or NF-κB-binding regions in VacA-induced IL-8 promoter activation. Thus, in U937 cells, VacA directly increases IL-8 production by activation of the p38 MAPK via intracellular Ca 2+ release, leading to activation of the transcription factors, ATF-2, CREB, and NF-κB.

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