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Infection and Immunity, March 2002, p. 1272-1278, Vol. 70, No. 3
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.3.1272-1278.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Human Gingival CD14+ Fibroblasts Primed with Gamma Interferon Increase Production of Interleukin-8 in Response to Lipopolysaccharide through Up-Regulation of Membrane CD14 and MyD88 mRNA Expression

Riyoko Tamai,1 Tetsuya Sakuta,2 Kenji Matsushita,2 Mitsuo Torii,2 Osamu Takeuchi,3 Shizuo Akira,3 Sachiko Akashi,4 Terje Espevik,5 Shunji Sugawara,1 and Haruhiko Takada1*

Department of Microbiology and Immunology, Tohoku University School of Dentistry, Sendai 980-8575,1 Department of Operative Dentistry and Endodontology, Kagoshima University Dental School, Kagoshima 890-8544,2 Department of Host Defense, Research Institute for Microbial Defenses, Osaka University, Osaka 565-0871,3 Division of Infectious Genetics, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan,4 Institute of Cancer Research and Molecular Biology, Norwegian University of Science and Technology, Trondheim, Norway5

Received 13 August 2001/ Returned for modification 26 September 2001/ Accepted 27 November 2001

Gamma interferon (IFN-{gamma})-primed human gingival fibroblasts (HGF) have been shown to produce higher levels of interleukin-8 (IL-8) upon stimulation with bacterial products and inflammatory cytokines than nonprimed controls. In this study, we examined whether priming of HGF with IFN-{gamma} up-regulates IL-8 production by the cells in response to purified lipopolysaccharide (LPS). The priming effect of IFN-{gamma} was clearly observed in the high-CD14-expressing (CD14high) HGF but not in the low-CD14-expressing (CD14low) HGF. The CD14high HGF were most effectively primed with IFN-{gamma} (1,000 IU/ml) for 72 h. To elucidate the mechanism of the priming effects of IFN-{gamma} for the LPS response by HGF, we examined whether IFN-{gamma} regulated expression of CD14, Toll-like receptor 2 (TLR2), TLR4, MD-2, and MyD88, all of which are molecules suggested to be associated with LPS signaling. In CD14high HGF, IFN-{gamma} markedly up-regulated CD14 and MyD88 but not TLR4 protein and MD-2 mRNA expression, while in CD14low HGF, IFN-{gamma} slightly increased MyD88 and scarcely affected CD14, TLR4 protein, and MD-2 mRNA levels. LPS-induced IL-8 production by IFN-{gamma}-primed CD14high HGF was significantly inhibited by monoclonal antibodies (MAbs) against CD14 and TLR4, but not by an anti-TLR2 MAb. These findings suggested that IFN-{gamma} primed CD14high HGF to enhance production of IL-8 in response to LPS through augmentation of the CD14-TLR system, where the presence of membrane CD14 was indispensable for the response of HGF to LPS.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Tohoku University School of Dentistry, 4-1 Seiryo-machi, Aoba-ku, Sendai 980-8575, Japan. Phone: 81 22 717 8305. Fax: 81 22 717 8309. E-mail: dent-ht{at}mail.cc.tohoku.ac.jp.

Infection and Immunity, March 2002, p. 1272-1278, Vol. 70, No. 3
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.3.1272-1278.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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