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Clinical and Diagnostic Laboratory Immunology, September 2004, p. 969-976, Vol. 11, No. 5
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.5.969-976.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Membrane-Anchored CD14 Is Important for Induction of Interleukin-8 by Lipopolysaccharide and Peptidoglycan in Uroepithelial Cells

Toshiaki Shimizu,1 Shin-ichi Yokota,2 Satoshi Takahashi,1 Yasuharu Kunishima,1 Koh Takeyama,1 Naoya Masumori,1 Atsushi Takahashi,1 Masanori Matsukawa,1 Naoki Itoh,1 Taiji Tsukamoto,1 and Nobuhiro Fujii2*

Department of Urology,1 Department of Microbiology, Sapporo Medical University School of Medicine, Chuo-ku, Sapporo, Japan2

Received 21 April 2004/ Returned for modification 3 June 2004/ Accepted 15 June 2004

We investigated the induction of interleukin-8 (IL-8) by bacterial lipopolysaccharide (LPS) and peptidoglycan (PGN) in the bladder cancer cell lines T24, 5637, UM-UC-3, and HT1197. T24 and 5637 cells strongly induced IL-8 after stimulation with LPS or PGN in a dose- and time-dependent manner, whereas UM-UC-3 and HT1197 cells did so very weakly. The expression of CD14 at the mRNA, total cellular protein, and cell surface protein levels differed among these cell lines, but the expression levels of Toll-like receptors 2 and 4 (TLR2 and TLR4) were not significantly different. The CD14 expression levels were found to correlate with the inducibility of IL-8 by LPS or PGN. Treatment of T24 and 5637 cells with phosphatidylinositol-specific phospholipase C to eliminate CD14 from the cell surface dramatically suppressed the induction of IL-8. On the other hand, UM-UC-3 cells transfected with CD14 cDNA expressed membrane-anchored CD14 and showed more efficent induction of IL-8 by LPS stimulation than untransfected controls. These results suggest that the presence of the membrane-anchored, but not the soluble, form of CD14 is a strong factor in IL-8 induction in bladder epithelial cells in response to bacterial components. The presence of the membrane-anchored form of CD14 may thus be a determinant for the inflammatory response of uroepithelial cells.

* Corresponding author. Mailing address: Department of Microbiology, Sapporo Medical University School of Medicine, South-1, West-17, Chuo-ku, Sapporo 060-8556, Hokkaido, Japan. Phone: 81-11-611-2111, ext. 2710. Fax: 81-11-612-5861. E-mail: fujii{at}sapmed.ac.jp.

Clinical and Diagnostic Laboratory Immunology, September 2004, p. 969-976, Vol. 11, No. 5
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.5.969-976.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.





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