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International Immunology Advance Access originally published online on November 6, 2007
International Immunology 2008 20(1):21-29; doi:10.1093/intimm/dxm115
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© The Japanese Society for Immunology. 2007. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

Involvement of phospholipase D in regulating expression of anti-microbial peptide human β-defensin-2

Suttichai Krisanaprakornkit1, Pareena Chotjumlong2, Prachya Kongtawelert2 and Vichai Reutrakul3

1 Department of Odontology and Oral Pathology, Faculty of Dentistry
2 Thailand Excellence Center for Tissue Engineering, Department of Biochemistry, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand
3 Department of Chemistry, Faculty of Science, Mahidol University, Bangkok 10400, Thailand

Correspondence to: V. Reutrakul; E-mail: scvrt{at}mahidol.ac.th

Human β-defensin expression correlates with differentiation in oral epithelium, and calcium ion, an important regulator of epithelial differentiation, plays a critical role in regulation of human β-defensin-2 (hBD-2) mRNA expression. Phospholipase D (PLD) also regulates epithelial differentiation. Therefore, we examined the role of PLD in hBD-2 up-regulation by cell wall extract of Fusobacterium nucleatum and phorbol 12-myristate 13-acetate (PMA), two known hBD-2 activators. We found that hBD-2 mRNA up-regulation in human gingival epithelial cells (HGECs) by these two activators was mediated by PLD activation and blocked by ethanol and 1-butanol, PLD inhibitors. PLD activity was induced by stimulation with these two activators, and phosphatidic acid (PA), a product generated from the PLD enzymatic activity, was detected in stimulated HGECs. Dioctanoyl PA commonly used for PA induced hBD-2 mRNA expression. mRNAs for PLD1{alpha} and β splice variants as well as PLD1 protein were constitutively expressed, whereas mRNA and protein for PLD2 were expressed at much lower levels than those for PLD1. Moreover, pre-treatment with (±)-propanolol, an inhibitor of phosphatidic acid phosphohydrolases that are the downstream signaling molecules in the PLD pathway, significantly blocked hBD-2 mRNA induction by PMA in a dose-dependent manner. In conclusion, these findings indicate the involvement of PLD activation in hBD-2 up-regulation in HGECs, which correlates with the state of epithelial differentiation.

Keywords: commensal bacteria, gene regulation, gingival epithelial cells, innate immunity, signal transduction

Transmitting editor: S. H. E. Haufman

Received 2 February 2007, accepted 11 October 2007.


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