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Infection and Immunity, July 2007, p. 3373-3381, Vol. 75, No. 7
0019-9567/07/$08.00+0     doi:10.1128/IAI.01940-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Vacuolating Cytotoxin in Helicobacter pylori Water-Soluble Proteins Upregulates Chemokine Expression in Human Eosinophils via Ca2+ Influx, Mitochondrial Reactive Oxygen Intermediates, and NF-{kappa}B Activation{triangledown}

Jung Mogg Kim,1* Joo Sung Kim,2 Jin Young Lee,1 Yeong-Jeon Kim,3 Ho-Joo Youn,4 In Young Kim,5 Young Joon Chee,5 Yu-Kyoung Oh,6 Nayoung Kim,2 Hyun Chae Jung,2 and In Sung Song2

Department of Microbiology, Hanyang University College of Medicine, Seoul 133-791, South Korea,1 Department of Internal Medicine and Liver Research Institute, Seoul National University College of Medicine, Seoul 110-744, South Korea,2 Department of Biotechnology, Joongbu University, Choongnam 312-940, South Korea,3 Department of Internal Medicine, Hanyang University College of Medicine, Seoul 133-791, South Korea,4 School of Life Sciences and Biotechnology, Korea University, Seoul 136-701, South Korea,5 Department of Biomedical Engineering, Hanyang University, Seoul 133-791, South Korea6

Received 11 December 2006/ Returned for modification 2 February 2007/ Accepted 9 April 2007

Helicobacter pylori-infected gastric mucosa is characterized by infiltration of inflammatory cells such as neutrophils and eosinophils. However, little information is available on the relationship between H. pylori virulence factors and chemokine expression in eosinophils. This study investigates the role of vacuolating cytotoxin (VacA) in chemokine expression from human eosinophils. Eosinophils were isolated from the peripheral blood of healthy volunteers using a magnetic cell separation system. VacA+ H. pylori water-soluble proteins (WSP) induced higher expression of interleukin-8, growth-related oncogene alpha, monocyte chemotactic protein 1, and RANTES (regulated on activation, normal, T-cell expressed and secreted) than Vac– WSP in human eosinophils, as assessed by quantitative reverse transcription-PCR and enzyme-linked immunosorbent assay. Purified VacA not only increased chemokine expression but also activated p65/p50 NF-{kappa}B heterodimers and phosphorylated I{kappa}B kinase (IKK) {alpha}/ß signals in human eosinophils. Inhibition of NF-{kappa}B and IKK significantly decreased the chemokine expression in VacA-stimulated eosinophils. Furthermore, VacA-induced NF-{kappa}B activation and chemokine release from eosinophils were dependent on Ca2+ influx and mitochondrial generation of reactive oxygen intermediates (ROI). These results suggest that NF-{kappa}B and IKK signals via Ca2+ influx and mitochondrial ROI play a role in the up-regulation of chemokine expression in eosinophils stimulated with H. pylori VacA.

* Corresponding author. Mailing address: Department of Microbiology, Hanyang University College of Medicine, 17 Haengdang-dong, Sungdong-gu, Seoul 133-791, South Korea. Phone: 82 2 2220 0645. Fax: 82 2 2282 0645. E-mail: jungmogg{at}hanyang.ac.kr

{triangledown} Published ahead of print on 23 April 2007.

Editor: F. C. Fang

Infection and Immunity, July 2007, p. 3373-3381, Vol. 75, No. 7
0019-9567/07/$08.00+0     doi:10.1128/IAI.01940-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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