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Essential role of MD-2 in LPS responsiveness and TLR4 distribution

Nature Immunology volume 3, pages 667–672 (2002)Cite this article

Abstract

Toll-like receptor 4 (TLR4) mediates lipopolysaccharide (LPS) signaling in a variety of cell types. MD-2 is associated with the extracellular domain of TLR4 and augments TLR4-dependent LPS responses in vitro. We show here that MD-2−/− mice do not respond to LPS, do survive endotoxic shock but are susceptible to Salmonella typhimurium infection. We found that in MD-2−/− embryonic fibroblasts, TLR4 was not able to reach the plasma membrane and predominantly resided in the Golgi apparatus, whereas TLR4 was distributed at the leading edge surface of cells in wild-type embryonic fibroblasts. Thus, MD-2 is essential for correct intracellular distribution and LPS-recognition of TLR4.

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Figure 1: Generation of MD-2−/− mice.
Figure 2: MD-2−/− B cells have impaired LPS responses.
Figure 3: LPS hyporesponsiveness of bone marrow–derived macrophages and DCs from MD-2−/− mice.
Figure 4: MD-2−/− mice have impaired LPS responses in vivo.
Figure 5: Intracellular distribution of TLR4 and organelle markers in MD-2−/− EF cells.

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Acknowledgements

We thank R. Horai, K. Fukudome and T. Furuta for technical suggestions; and P. W. Kincade and D. R. Liddicoat for helpful comments on the manuscript. Supported by Special Coordination Funds of the Ministry of Education, Culture, Sports, Science and Technology of the Japanese Government (Monbukagakusho); the Uehara Memorial Foundation; the Yamanouchi Foundation for Research on Metabolic Disorders; Mitsubishi Pharma; and Sankyo.

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Authors and Affiliations

  1. Division of Infectious Genetics, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan

    Yoshinori Nagai, Sachiko Akashi & Kensuke Miyake

  2. CREST, Japan Science and Technology Corporation, Tokyo, Japan

    Yoshinori Nagai, Sachiko Akashi, Masakazu Nagafuku, Atsushi Kosugi & Kensuke Miyake

  3. Department of Immunology, Saga Medical School, Saga, Japan

    Yoshinori Nagai & Masao Kimoto

  4. School of Allied Health Sciences, Faculty of Medicine, Osaka, Japan

    Masakazu Nagafuku & Atsushi Kosugi

  5. Department of Oncogenesis, Osaka University Medical School, Osaka, Japan

    Masato Ogata

  6. The Center for Experimental Medicine, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan

    Yoichiro Iwakura

  7. Research Institute for Microbial Diseases, Osaka University, Japan

    Shizuo Akira

  8. Division of Cellular Therapy, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan

    Toshio Kitamura

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Correspondence to Kensuke Miyake.

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Nagai, Y., Akashi, S., Nagafuku, M. et al. Essential role of MD-2 in LPS responsiveness and TLR4 distribution. Nat Immunol 3, 667–672 (2002). https://doi.org/10.1038/ni809

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