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Infection and Immunity, May 2001, p. 2788-2796, Vol. 69, No. 5
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.5.2788-2796.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Upregulation of Toll-Like Receptor 2 Gene Expression in Macrophage Response to Peptidoglycan and High Concentration of Lipopolysaccharide Is Involved in NF-kappa B Activation

Yan Liu,1 Yin Wang,1 Munekazu Yamakuchi,1 Sumikazu Isowaki,2 Etsuro Nagata,2 Yuichi Kanmura,2 Isao Kitajima,1 and Ikuro Maruyama1,*

Department of Laboratory and Molecular Medicine1 and Department of Anesthesiology and Critical Care Medicine, Faculty of Medicine,2 Kagoshima University, 8-35-1 Sakuragaoka, Kagoshima 890-8520, Japan

Received 8 September 2000/Returned for modification 8 November 2000/Accepted 24 January 2001

Toll-like receptors 2 and 4 (TLR2 and TLR4) have been found to transduce signals of peptidoglycan (PGN) and lipopolysaccharide (LPS), respectively, for NF-kappa B activation. However, little is known about the expression and regulation of the TLR2 gene in monocytes/macrophages in response to the two typical bacterial products. We show in the present study that both PGN and a high concentration of LPS increase TLR2 gene expression in macrophage-like cells, 1alpha ,25-dihydroxyvitamin D3-differentiated human HL60 and mouse RAW264.7 cells, and human monocytes in a dose- and time-dependent manner. Actinomycin D and pyrrolidine dithiocarbamate inhibition of gene transcription and NF-kappa B activation, respectively, blocks LPS- and PGN-elevated TLR2 mRNA in monocytic cells. The LPS-induced increase in TLR2 mRNA in monocytic cells is abolished by polymyxin B pretreatment and is observed in peripheral blood mononuclear cells from pigs subjected to endotoxic shock. Further, high concentrations of LPS and synthetic lipid A increase TLR2 mRNA expression in peritoneal macrophages from both TLR4-deficient C3H/HeJ mice and normal C3H/HeN mice, a process that constitutes induction of TLR4-independent TLR2 expression. These findings demonstrate that TLR2 gene expression is upregulated in macrophage responses to PGN and to high concentrations of LPS in vitro and in vivo and correlates with NF-kappa B activation.

* Corresponding author. Mailing address: Department of Laboratory and Molecular Medicine, Kagoshima University School of Medicine, 8-35-1 Sakuragaoka, Kagoshima City 890-8520, Japan. Phone: 81 (99) 275-5437. Fax: 81 (99) 275-2629. E-mail: rinken1{at}khosp2.kufm.kagoshima-u.ac.jp.

Infection and Immunity, May 2001, p. 2788-2796, Vol. 69, No. 5
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.5.2788-2796.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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