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-kappaB 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

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-kappaB 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

Department of Biomedical Sciences

76 Citations (Scopus)

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), IkappaBalpha ((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-activated p38 MAPK, caused a dose-dependent reduction in VacA-induced IL-8 secretion by U937 cells, implying a role for intracellular Ca(2+) in mediating activation of MAPK and the canonical NF-kappaB pathway. VacA stimulated translocation of NF-kappaBp65 to the nucleus, consistent with enhancement of IL-8 expression by activation of the NF-kappaB 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-kappaB site, suggesting direct involvement of the ATF-2/CREB binding region or NF-kappaB-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-kappaB.

Original language	English
Journal	Journal of Immunology
Volume	180
Issue number	7
Pages (from-to)	5017-27
Number of pages	10
ISSN	0022-1767
Publication status	Published - 2008

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Hisatsune, J., Nakayama, M., Isomoto, H., Kurazono, H., Mukaida, N., Mukhopadhyay, A. K., 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-kappaB activation. Journal of Immunology, 180(7), 5017-27.

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-kappaB activation. / Hisatsune, Junzo; Nakayama, Masaaki; Isomoto, Hajime et al.

In: Journal of Immunology, Vol. 180, No. 7, 2008, p. 5017-27.

Research output: Contribution to journal › Journal article › Research › peer-review

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-kappaB activation.', Journal of Immunology, vol. 180, no. 7, pp. 5017-27.

@article{b683faf054a611dd8d9f000ea68e967b,

title = "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-kappaB activation.",

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), IkappaBalpha ((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-activated p38 MAPK, caused a dose-dependent reduction in VacA-induced IL-8 secretion by U937 cells, implying a role for intracellular Ca(2+) in mediating activation of MAPK and the canonical NF-kappaB pathway. VacA stimulated translocation of NF-kappaBp65 to the nucleus, consistent with enhancement of IL-8 expression by activation of the NF-kappaB 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-kappaB site, suggesting direct involvement of the ATF-2/CREB binding region or NF-kappaB-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-kappaB.",

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

note = "Keywords: Activating Transcription Factor 2; Active Transport, Cell Nucleus; Bacterial Proteins; Butylated Hydroxyanisole; Cell Nucleus; Cyclic AMP Response Element-Binding Protein; Dantrolene; Egtazic Acid; Enzyme Activation; Helicobacter pylori; Humans; Imidazoles; Interleukin-8; NF-kappa B; Nitriles; Phosphorylation; Promoter Regions (Genetics); Sulfones; Thapsigargin; U937 Cells; Up-Regulation; p38 Mitogen-Activated Protein Kinases",

year = "2008",

language = "English",

volume = "180",

pages = "5017--27",

journal = "Journal of Immunology",

issn = "0022-1767",

publisher = "American Association of Immunologists",

number = "7",

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TY - JOUR

T1 - 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-kappaB activation.

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

N1 - Keywords: Activating Transcription Factor 2; Active Transport, Cell Nucleus; Bacterial Proteins; Butylated Hydroxyanisole; Cell Nucleus; Cyclic AMP Response Element-Binding Protein; Dantrolene; Egtazic Acid; Enzyme Activation; Helicobacter pylori; Humans; Imidazoles; Interleukin-8; NF-kappa B; Nitriles; Phosphorylation; Promoter Regions (Genetics); Sulfones; Thapsigargin; U937 Cells; Up-Regulation; p38 Mitogen-Activated Protein Kinases

PY - 2008

Y1 - 2008

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), IkappaBalpha ((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-activated p38 MAPK, caused a dose-dependent reduction in VacA-induced IL-8 secretion by U937 cells, implying a role for intracellular Ca(2+) in mediating activation of MAPK and the canonical NF-kappaB pathway. VacA stimulated translocation of NF-kappaBp65 to the nucleus, consistent with enhancement of IL-8 expression by activation of the NF-kappaB 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-kappaB site, suggesting direct involvement of the ATF-2/CREB binding region or NF-kappaB-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-kappaB.

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), IkappaBalpha ((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-activated p38 MAPK, caused a dose-dependent reduction in VacA-induced IL-8 secretion by U937 cells, implying a role for intracellular Ca(2+) in mediating activation of MAPK and the canonical NF-kappaB pathway. VacA stimulated translocation of NF-kappaBp65 to the nucleus, consistent with enhancement of IL-8 expression by activation of the NF-kappaB 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-kappaB site, suggesting direct involvement of the ATF-2/CREB binding region or NF-kappaB-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-kappaB.

M3 - Journal article

C2 - 18354227

SN - 0022-1767

VL - 180

SP - 5017

EP - 5027

JO - Journal of Immunology

JF - Journal of Immunology

IS - 7

ER -