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The Journal of Immunology, 2000, 164: 3471-3475.
Copyright © 2000 by The American Association of Immunologists

CUTTING EDGE

Cutting Edge: Cell Surface Expression and Lipopolysaccharide Signaling Via the Toll-Like Receptor 4-MD-2 Complex on Mouse Peritoneal Macrophages1

Sachiko Akashi*, Rintaro Shimazu*, Hirotaka Ogata*, Yoshinori Nagai*, Kiyoshi Takeda{dagger}, Masao Kimoto* and Kensuke Miyake2,*

* Department of Immunology, Saga Medical School, Nabeshima, Japan; and {dagger} Department of Host Defense, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan

The human MD-2 molecule is associated with the extracellular domain of human Toll-like receptor 4 (TLR4) and greatly enhances its LPS signaling. The human TLR4-MD-2 complex thus signals the presence of LPS. Little is known, however, about cell surface expression and LPS signaling of the TLR4-MD-2 complex in vivo. We cloned mouse MD-2 molecularly and established a unique mAb MTS510, which reacted selectively with mouse TLR4-MD-2 but not with TLR4 alone in flow cytometry. Mouse MD-2 expression in TLR4-expressing cells enhanced LPS-induced NF-{kappa}B activation, which was clearly inhibited by MTS510. Thioglycolate-elicited peritoneal macrophages expressed TLR4-MD-2, which was rapidly down-regulated in the presence of LPS. Moreover, LPS-induced TNF-{alpha} production by peritoneal macrophages was inhibited by MTS510. Collectively, the TLR4-MD-2 complex is expressed on macrophages in vivo and senses and signals the presence of LPS.




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F. Re and J. L. Strominger
Separate Functional Domains of Human MD-2 Mediate Toll-Like Receptor 4-Binding and Lipopolysaccharide Responsiveness
J. Immunol., November 15, 2003; 171(10): 5272 - 5276.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
M. Hashimoto, Y. Asai, and T. Ogawa
Treponemal Phospholipids Inhibit Innate Immune Responses Induced by Pathogen-associated Molecular Patterns
J. Biol. Chem., November 7, 2003; 278(45): 44205 - 44213.
[Abstract] [Full Text] [PDF]

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 JEMHome page
M. W. Hornef, B. H. Normark, A. Vandewalle, and S. Normark
Intracellular Recognition of Lipopolysaccharide by Toll-like Receptor 4 in Intestinal Epithelial Cells
J. Exp. Med., October 20, 2003; 198(8): 1225 - 1235.
[Abstract] [Full Text] [PDF]

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 JEMHome page
S. Akashi, S.-i. Saitoh, Y. Wakabayashi, T. Kikuchi, N. Takamura, Y. Nagai, Y. Kusumoto, K. Fukase, S. Kusumoto, Y. Adachi, et al.
Lipopolysaccharide Interaction with Cell Surface Toll-like Receptor 4-MD-2: Higher Affinity than That with MD-2 or CD14
J. Exp. Med., October 6, 2003; 198(7): 1035 - 1042.
[Abstract] [Full Text] [PDF]

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 Innate ImmunityHome page
K. Kawasaki, K. Gomi, Y. Kawai, M. Shiozaki, and M. Nishijima
Molecular basis for lipopolysaccharide mimetic action of TaxolTM and flavolipin
Innate Immunity, October 1, 2003; 9(5): 301 - 307.
[Abstract] [PDF]

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 J. Immunol.Home page
S. C. Higgins, E. C. Lavelle, C. McCann, B. Keogh, E. McNeela, P. Byrne, B. O'Gorman, A. Jarnicki, P. McGuirk, and K. H. G. Mills
Toll-Like Receptor 4-Mediated Innate IL-10 Activates Antigen-Specific Regulatory T Cells and Confers Resistance to Bordetella pertussis by Inhibiting Inflammatory Pathology
J. Immunol., September 15, 2003; 171(6): 3119 - 3127.
[Abstract] [Full Text] [PDF]

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 J. Immunol.Home page
H. M. Paterson, T. J. Murphy, E. J. Purcell, O. Shelley, S. J. Kriynovich, E. Lien, J. A. Mannick, and J. A. Lederer
Injury Primes the Innate Immune System for Enhanced Toll-Like Receptor Reactivity
J. Immunol., August 1, 2003; 171(3): 1473 - 1483.
[Abstract] [Full Text] [PDF]

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 J. Immunol.Home page
N. Qureshi, P.-Y. Perera, J. Shen, G. Zhang, A. Lenschat, G. Splitter, D. C. Morrison, and S. N. Vogel
The Proteasome as a Lipopolysaccharide-Binding Protein in Macrophages: Differential Effects of Proteasome Inhibition on Lipopolysaccharide-Induced Signaling Events
J. Immunol., August 1, 2003; 171(3): 1515 - 1525.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Cell Physiol.Home page
S. Okugawa, Y. Ota, T. Kitazawa, K. Nakayama, S. Yanagimoto, K. Tsukada, M. Kawada, and S. Kimura
Janus kinase 2 is involved in lipopolysaccharide-induced activation of macrophages
Am J Physiol Cell Physiol, August 1, 2003; 285(2): C399 - C408.
[Abstract] [Full Text] [PDF]

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 J. Immunol.Home page
C. E. O. Baleeiro, S. E. Wilcoxen, S. B. Morris, T. J. Standiford, and R. Paine III
Sublethal Hyperoxia Impairs Pulmonary Innate Immunity
J. Immunol., July 15, 2003; 171(2): 955 - 963.
[Abstract] [Full Text] [PDF]

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 J. Leukoc. Biol.Home page
D. Dory, H. Echchannaoui, M. Letiembre, F. Ferracin, J. Pieters, Y. Adachi, S. Akashi, W. Zimmerli, and R. Landmann
Generation and functional characterization of a clonal murine periportal Kupffer cell line from H-2Kb -tsA58 mice
J. Leukoc. Biol., July 1, 2003; 74(1): 49 - 59.
[Abstract] [Full Text] [PDF]

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 J. Immunol.Home page
F. Bihl, L. Salez, M. Beaubier, D. Torres, L. Lariviere, L. Laroche, A. Benedetto, D. Martel, J.-M. Lapointe, B. Ryffel, et al.
Overexpression of Toll-Like Receptor 4 Amplifies the Host Response to Lipopolysaccharide and Provides a Survival Advantage in Transgenic Mice
J. Immunol., June 15, 2003; 170(12): 6141 - 6150.
[Abstract] [Full Text] [PDF]

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 Innate ImmunityHome page
R. Tamai, S. Sugawara, O. Takeuchi, S. Akira, and H. Takada
Synergistic effects of lipopolysaccharide and interferon-{gamma} in inducing interleukin-8 production in human monocytic THP-1 cells is accompanied by up-regulation of CD14, Toll-like receptor 4, MD-2 and MyD88 expression
Innate Immunity, June 1, 2003; 9(3): 145 - 153.
[Abstract] [PDF]

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 J. Biol. Chem.Home page
T. Pedron, R. Girard, and R. Chaby
TLR4-dependent Lipopolysaccharide-induced Shedding of Tumor Necrosis Factor Receptors in Mouse Bone Marrow Granulocytes
J. Biol. Chem., May 30, 2003; 278(23): 20555 - 20564.
[Abstract] [Full Text] [PDF]

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 CVIHome page
T. Ohnishi, M. Muroi, and K.-i. Tanamoto
MD-2 Is Necessary for the Toll-Like Receptor 4 Protein To Undergo Glycosylation Essential for Its Translocation to the Cell Surface
Clin. Vaccine Immunol., May 1, 2003; 10(3): 405 - 410.
[Abstract] [Full Text] [PDF]

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 J. Immunol.Home page
C. F. Ortega-Cava, S. Ishihara, M. A. K. Rumi, K. Kawashima, N. Ishimura, H. Kazumori, J. Udagawa, Y. Kadowaki, and Y. Kinoshita
Strategic Compartmentalization of Toll-Like Receptor 4 in the Mouse Gut
J. Immunol., April 15, 2003; 170(8): 3977 - 3985.
[Abstract] [Full Text] [PDF]

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 Innate ImmunityHome page
T. Roger, C. Froidevaux, C. Martin, and T. Calandra
Macrophage migration inhibitory factor (MIF) regulates host responses to endotoxin through modulation of Toll-like receptor 4 (TLR4)
Innate Immunity, April 1, 2003; 9(2): 119 - 123.
[Abstract] [PDF]

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 JNCI J Natl Cancer InstHome page
M. Okamoto, T. Oshikawa, T. Tano, G. Ohe, S. Furuichi, H. Nishikawa, S. U. Ahmed, S. Akashi, K. Miyake, O. Takeuchi, et al.
Involvement of Toll-Like Receptor 4 Signaling in Interferon-{gamma} Production and Antitumor Effect by Streptococcal Agent OK-432
J Natl Cancer Inst, February 19, 2003; 95(4): 316 - 326.
[Abstract] [Full Text] [PDF]

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 JEMHome page
I. Caramalho, T. Lopes-Carvalho, D. Ostler, S. Zelenay, M. Haury, and J. Demengeot
Regulatory T Cells Selectively Express Toll-like Receptors and Are Activated by Lipopolysaccharide
J. Exp. Med., February 17, 2003; 197(4): 403 - 411.
[Abstract] [Full Text] [PDF]

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 Infect. Immun.Home page
L. A. Augusto, M. Synguelakis, J. Johansson, T. Pedron, R. Girard, and R. Chaby
Interaction of Pulmonary Surfactant Protein C with CD14 and Lipopolysaccharide
Infect. Immun., January 1, 2003; 71(1): 61 - 67.
[Abstract] [Full Text] [PDF]

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 J. Immunol.Home page
K. Kawasaki, H. Nogawa, and M. Nishijima
Identification of Mouse MD-2 Residues Important for Forming the Cell Surface TLR4-MD-2 Complex Recognized by Anti-TLR4-MD-2 Antibodies, and for Conferring LPS and Taxol Responsiveness on Mouse TLR4 by Alanine-Scanning Mutagenesis
J. Immunol., January 1, 2003; 170(1): 413 - 420.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
T. L. Gioannini, D. Zhang, A. Teghanemt, and J. P. Weiss
An Essential Role for Albumin in the Interaction of Endotoxin with Lipopolysaccharide-binding Protein and sCD14 and Resultant Cell Activation
J. Biol. Chem., November 27, 2002; 277(49): 47818 - 47825.
[Abstract] [Full Text] [PDF]

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 Am. J. Pathol.Home page
K. Yamamoto, T. Shimokawa, H. Yi, K.-i. Isobe, T. Kojima, D. J. Loskutoff, and H. Saito
Aging Accelerates Endotoxin-Induced Thrombosis : Increased Responses of Plasminogen Activator Inhibitor-1 and Lipopolysaccharide Signaling with Aging
Am. J. Pathol., November 1, 2002; 161(5): 1805 - 1814.
[Abstract] [Full Text] [PDF]

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 J. Immunol.Home page
N. Tsuboi, Y. Yoshikai, S. Matsuo, T. Kikuchi, K.-I. Iwami, Y. Nagai, O. Takeuchi, S. Akira, and T. Matsuguchi
Roles of Toll-Like Receptors in C-C Chemokine Production by Renal Tubular Epithelial Cells
J. Immunol., August 15, 2002; 169(4): 2026 - 2033.
[Abstract] [Full Text] [PDF]

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 J Med MicrobiolHome page
A. UEHARA, S. SUGAWARA, and H. TAKADA
Priming of human oral epithelial cells by interferon-{gamma} to secrete cytokines in response to lipopolysaccharides, lipoteichoic acids and peptidoglycans
J. Med. Microbiol., August 1, 2002; 51(8): 626 - 634.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
S. B. Mizel and J. A. Snipes
Gram-negative Flagellin-induced Self-tolerance Is Associated with a Block in Interleukin-1 Receptor-associated Kinase Release from Toll-like Receptor 5
J. Biol. Chem., June 14, 2002; 277(25): 22414 - 22420.
[Abstract] [Full Text] [PDF]

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 J. Immunol.Home page
J. Fan, A. Kapus, P. A. Marsden, Y. H. Li, G. Oreopoulos, J. C. Marshall, S. Frantz, R. A. Kelly, R. Medzhitov, and O. D. Rotstein
Regulation of Toll-Like Receptor 4 Expression in the Lung Following Hemorrhagic Shock and Lipopolysaccharide
J. Immunol., May 15, 2002; 168(10): 5252 - 5259.
[Abstract] [Full Text] [PDF]

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 Infect. Immun.Home page
S. Li, V. M. Holers, S. A. Boackle, and C. M. Blatteis
Modulation of Mouse Endotoxic Fever by Complement
Infect. Immun., May 1, 2002; 70(5): 2519 - 2525.
[Abstract] [Full Text] [PDF]

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 IOVSHome page
N. Tuaillon, D. F. Shen, R. B. Berger, B. Lu, B. J. Rollins, and C.-C. Chan
MCP-1 Expression in Endotoxin-Induced Uveitis
Invest. Ophthalmol. Vis. Sci., May 1, 2002; 43(5): 1493 - 1498.
[Abstract] [Full Text] [PDF]

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 J. Immunol.Home page
K. Dabbagh, M. E. Dahl, P. Stepick-Biek, and D. B. Lewis
Toll-Like Receptor 4 Is Required for Optimal Development of Th2 Immune Responses: Role of Dendritic Cells
J. Immunol., May 1, 2002; 168(9): 4524 - 4530.
[Abstract] [Full Text] [PDF]

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 J. Immunol.Home page
K. Gomi, K. Kawasaki, Y. Kawai, M. Shiozaki, and M. Nishijima
Toll-Like Receptor 4-MD-2 Complex Mediates the Signal Transduction Induced by Flavolipin, an Amino Acid-Containing Lipid Unique to Flavobacterium meningosepticum
J. Immunol., March 15, 2002; 168(6): 2939 - 2943.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
J. J. Watters, J. A. Sommer, Z. A. Pfeiffer, U. Prabhu, A. N. Guerra, and P. J. Bertics
A Differential Role for the Mitogen-activated Protein Kinases in Lipopolysaccharide Signaling. THE MEK/ERK PATHWAY IS NOT ESSENTIAL FOR NITRIC OXIDE AND INTERLEUKIN 1beta PRODUCTION
J. Biol. Chem., March 8, 2002; 277(11): 9077 - 9087.
[Abstract] [Full Text] [PDF]

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 Infect. Immun.Home page
T. Vasselon and P. A. Detmers
Toll Receptors: a Central Element in Innate Immune Responses
Infect. Immun., March 1, 2002; 70(3): 1033 - 1041.
[Full Text] [PDF]

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 Infect. Immun.Home page
R. Tamai, T. Sakuta, K. Matsushita, M. Torii, O. Takeuchi, S. Akira, S. Akashi, T. Espevik, S. Sugawara, and H. Takada
Human Gingival CD14+ Fibroblasts Primed with Gamma Interferon Increase Production of Interleukin-8 in Response to Lipopolysaccharide through Up-Regulation of Membrane CD14 and MyD88 mRNA Expression
Infect. Immun., March 1, 2002; 70(3): 1272 - 1278.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
N. Iovine, J. Eastvold, P. Elsbach, J. P. Weiss, and T. L. Gioannini
The Carboxyl-terminal Domain of Closely Related Endotoxin-binding Proteins Determines the Target of Protein-Lipopolysaccharide Complexes
J. Biol. Chem., March 1, 2002; 277(10): 7970 - 7978.
[Abstract] [Full Text] [PDF]

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 J. Immunol.Home page
R. N. Fichorova, A. O. Cronin, E. Lien, D. J. Anderson, and R. R. Ingalls
Response to Neisseria gonorrhoeae by Cervicovaginal Epithelial Cells Occurs in the Absence of Toll-Like Receptor 4-Mediated Signaling
J. Immunol., March 1, 2002; 168(5): 2424 - 2432.
[Abstract] [Full Text] [PDF]

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 JEMHome page
M. W. Hornef, T. Frisan, A. Vandewalle, S. Normark, and A. Richter-Dahlfors
Toll-like Receptor 4 Resides in the Golgi Apparatus and Colocalizes with Internalized Lipopolysaccharide in Intestinal Epithelial Cells
J. Exp. Med., February 25, 2002; 195(5): 559 - 570.
[Abstract] [Full Text] [PDF]


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