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Infection and Immunity, August 2001, p. 4951-4957, Vol. 69, No. 8
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.8.4951-4957.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Monocytic Cell Activation by Nonendotoxic Glycoprotein from Prevotella intermedia ATCC 25611 Is Mediated by Toll-Like Receptor 2

Shunji Sugawara,1,* Shuhua Yang,1 Koichi Iki,2 Junko Hatakeyama,1,3 Riyoko Tamai,1 Osamu Takeuchi,4 Sachiko Akashi,5 Terje Espevik,6 Shizuo Akira,4 and Haruhiko Takada1

Department of Microbiology and Immunology1 and Department of Operative Dentistry,3 Tohoku University School of Dentistry, Sendai 980-8575, Department of Periodontology, Kagoshima University Dental School, Kagoshima 890-8544,2 Department of Host Defense, Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871,4 and Department of Immunology, Saga Medical School, Saga 849-8501,5 Japan, and Institute of Cancer Research and Molecular Biology, Norwegian University of Science and Technology, Trondheim, Norway6

Received 28 December 2000/Returned for modification 30 March 2001/Accepted 29 April 2001

Lipopolysaccharide (LPS) preparations from gram-negative black-pigmented bacteria such as Porphyromonas gingivalis and Prevotella intermedia activate cells from non-LPS-responsive C3H/HeJ mice, but it is still unclear whether this activity is due to the unique structure of LPS or to a minor component(s) responsible for the activity in the preparation. A nonendotoxic glycoprotein with bioactivity against cells from C3H/HeJ mice was purified from a hot phenol-water extract of P. intermedia ATCC 25611 and designated Prevotella glycoprotein (PGP). Treatment of human monocytic THP-1 cells with 22-oxyacalcitriol (OCT) induced maturation and marked expression of CD14 on the cells, but the cells constitutively expressed Toll-like receptor 2 (TLR2) and TLR4 on the cells irrespective of the treatment. PGP induced a high level of interleukin-8 production at doses of 100 ng/ml and higher in OCT-treated THP-1 cells compared with Salmonella LPS, and the production was significantly inhibited by anti-CD14 and anti-TLR2 but not anti-TLR4 antibodies. Consistent with this, TLR2-deficient murine macrophages did not respond to PGP. It was also shown that PGP activity on the THP-1 cells was LPS-binding protein dependent and was inhibited by a synthetic lipid A precursor IVA. These results indicate that PGP activates monocytic cells in a CD14- and TLR2-dependent manner.

* 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: 81-22-717-8306. Fax: 81-22-717-8309. E-mail: sugawars{at}mail.cc.tohoku.ac.jp.

Infection and Immunity, August 2001, p. 4951-4957, Vol. 69, No. 8
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.8.4951-4957.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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