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Infection and Immunity, August 2005, p. 4634-4642, Vol. 73, No. 8
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.8.4634-4642.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Toll-Like Receptor 4 Mediates Tolerance in Macrophages Stimulated with Toxoplasma gondii-Derived Heat Shock Protein 70

Hye-Seong Mun,1 Fumie Aosai,1 Kazumi Norose,1 Lian-Xun Piao,1 Hao Fang,1 Shizuo Akira,2 and Akihiko Yano1*

Department of Infection and Host Defense, Graduate School of Medicine, Chiba University, Chuo-ku, Chiba 260-8670,1 Department of Host Defense, Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871, Japan2

Received 16 November 2004/ Returned for modification 3 January 2005/ Accepted 14 March 2005

Peritoneal macrophages (PMs) from toll-like receptor 4 (TLR4)-deficient and wild-type (WT) mice were responsive to recombinant Toxoplasma gondii-derived heat shock protein 70 (rTgHSP70) and natural TgHSP70 (nTgHSP70) in NO release, but those from TLR2-, myeloid differentiation factor 88 (MyD88)-, and interleukin-1R-associated kinase 4 (IRAK4)-deficient mice were not. Polymyxin B did not inhibit PM activation by TgHSP70 and nTgHSP70 from WT and TLR4-deficient mice, while it inhibited PM activation by lipopolysaccharide. Pretreatment of PMs from WT but not from TLR4-deficient mice with rTgHSP70 resulted in suppression of NO release on restimulation with rTgHSP70. Similarly, pretreatment of PMs from WT but not TLR4-deficient mice with nTgHSP70 resulted in suppression of NO release on restimulation with nTgHSP70. Polymyxin B did not inhibit rTgHSP70- and nTgHSP70-induced tolerance of PMs from TLR4-deficient mice. Furthermore, PMs from WT mice increased suppressor of cytokine-signaling-1 (SOCS-1) expression after restimulation with rTgHSP70, while those from TLR4-deficient mice did not. Phosphorylation of JNK and I-{kappa}B{alpha} occurred in rTgHSP70-induced tolerance of PMs from TLR4-deficient mice, but not in that from WT mice. These data indicated that TgHSP70 signaling mechanisms were mediated by TLR2, MyD88, and IRAK4, but not by TLR4. On the other hand, signaling of TgHSP70-induced tolerance was mediated by TLR4, and the expression of SOCS-1 suppressed the TLR2 signaling pathway.

* Corresponding author. Mailing address: Department of Infection and Host Defense, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan. Phone: (81)-43-226-2071. Fax: (81)-43-226-2076. E-mail: yano{at}faculty.chiba-u.jp.

Editor: J. F. Urban, Jr.

Infection and Immunity, August 2005, p. 4634-4642, Vol. 73, No. 8
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.8.4634-4642.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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