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Infection and Immunity, June 2003, p. 3221-3226, Vol. 71, No. 6
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.6.3221-3226.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Lipopolysaccharide-Mimetic Activities of a Toll-Like Receptor 2-Stimulatory Substance(s) in Enterobacterial Lipopolysaccharide Preparations

Masashi Muroi, Takahiro Ohnishi, Satoko Azumi-Mayuzumi, and Ken-ichi Tanamoto*

Division of Microbiology, National Institute of Health Sciences, Setagaya, Tokyo 158-8501, Japan

Received 4 November 2002/ Returned for modification 18 February 2003/ Accepted 12 March 2003

Lipopolysaccharide (LPS) preparations are known to often contain substances which activate cells through Toll-like receptor 2 (TLR2), and it is suspected that bacterial lipoproteins are responsible for this activation. We compared the mode of action of the TLR2-stimulatory substances with that of a synthetic bacterial lipopeptide (tripalmitoyl-Cys-Ser-Ser-Asn-Ala [Pam3CSSNA]), as well as with that of peptidoglycan. Six out of eight LPS preparations tested induced NF-{kappa}B-dependent reporter activity in 293 cells expressing CD14 and TLR2. Phenol extract (PEX) prepared from Escherichia coli LPS by modified phenol extraction induced reporter activity in 293 cells expressing TLR2, and this activity was enhanced by coexpression of CD14, whereas the activity of Pam3CSSNA was not dependent on CD14. The activity of PEX, but not that of Pam3CSSNA or peptidoglycan, was also enhanced by LPS binding protein or serum and blocked by polymyxin B. In addition, the activity of PEX was inhibited by a lipid A precursor (compound 406) in 293 cells expressing CD14 and TLR2. These results indicate that E. coli LPS preparations contain LPS-mimetic TLR2-stimulatory substances which differ from bacterial lipopeptides or peptidoglycan.

* Corresponding author. Mailing address: Division of Microbiology, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya, Tokyo 158-8501, Japan. Phone: 81-3-3700-1141, ext. 272. Fax: 81-3-3707-6950. E-mail: tanamoto{at}nihs.go.jp.

Editor: A. D. O'Brien

Infection and Immunity, June 2003, p. 3221-3226, Vol. 71, No. 6
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.6.3221-3226.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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