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J. Biol. Chem., Vol. 277, Issue 49, 47834-47843, December 6, 2002
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From the Mammalian responses to LPS require the expression
of Toll-like receptor 4 (TLR4), CD14, and MD-2. We expressed
fluorescent TLR4 in cell lines and found that TLR4 densely localized to
the surface and the Golgi. Similar distributions were observed in human
monocytes. Confocal imaging revealed rapid recycling of TLR4-CD14-MD-2
complexes between the Golgi and the plasma membrane. Fluorescent LPS
followed these trafficking pathways in CD14-positive cells. The
TLR4- adapter protein, MyD88, translocated to the cell surface upon
LPS exposure, and cross-linking of surface TLR4 with antibody induced
signaling. Golgi-associated TLR4 expression was disrupted by brefeldin
A, yet LPS signaling was preserved. We conclude that LPS signaling may
be initiated by surface aggregation of TLR4 and is not dependent upon
LPS trafficking to the Golgi.
Lipopolysaccharide Rapidly Traffics to and
from the Golgi Apparatus with the Toll-like Receptor
4-MD-2-CD14 Complex in a Process That Is Distinct from the
Initiation of Signal Transduction*,
§,,
,,,,¶, and**
University of Massachusetts Medical School,
Division of Infectious Diseases, Worcester, Massachusetts 01605 and the
Norwegian University of Science and Technology,
Trondheim N-7489, Norway
*
This work was supported by National Institutes of Health
Grants GM54060, GM63244, and DK50305 (to D. T. G.), the Commission of
the European Communities, specific RTD program "Quality of Life and
Management of Living Resources" Grant QLK2-2000-336, HOSPATH,
the Norwegian Cancer Society, and the Norwegian Research Council (to
T. E.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
The on-line version of this article (available at
http://www.jbc.org) contains two videos. Video 1 shows CD14 movement
inside HEK cells stably transfected with CD14. A Tricolor-conjugated anti-CD14 antibody was used to stain CD14. Video 2 shows LPS uptake together with CD14. HEK-TLR4-CD14 cells were incubated with
Tricolor-conjugated anti-CD14 antibody (red) and BODIPY-LPS (green).
Shown are sequential images acquired by time-lapse confocal imaging in
the order channel red, channel green, and the overlay of red and green.
§
Supported by a stipend from the German Academic Exchange Program.
¶
These authors contributed equally to this work.
**
To whom correspondence should be addressed: Institute of Cancer
Research and Molecular Biology, Norwegian University of Science and
Technology, N-7489 Trondheim, Norway. Tel.: 47-73598668; Fax: 47-73598801; E-mail: terje.espevik@medisin.ntnu.no.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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 E Cario BACTERIAL INTERACTIONS WITH CELLS OF THE INTESTINAL MUCOSA: TOLL-LIKE RECEPTORS AND NOD2 Gut, August 1, 2005; 54(8): 1182 - 1193. [Full Text] [PDF]
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H. An, H. Xu, M. Zhang, J. Zhou, T. Feng, C. Qian, R. Qi, and X. Cao Src homology 2 domain-containing inositol-5-phosphatase 1 (SHIP1) negatively regulates TLR4-mediated LPS response primarily through a phosphatase activity- and PI-3K-independent mechanism Blood, June 15, 2005; 105(12): 4685 - 4692. [Abstract] [Full Text] [PDF]
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G. Tunheim, K. W. Schjetne, A. B. Fredriksen, I. Sandlie, and B. Bogen Human CD14 is an efficient target for recombinant immunoglobulin vaccine constructs that deliver T cell epitopes J. Leukoc. Biol., March 1, 2005; 77(3): 303 - 310. [Abstract] [Full Text] [PDF]
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M. Triantafilou and K. Triantafilou Invited review: The dynamics of LPS recognition: complex orchestration of multiple receptors Innate Immunity, February 1, 2005; 11(1): 5 - 11. [Abstract] [PDF]
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E. Aksoy, C. S. Zouain, F. Vanhoutte, J. Fontaine, N. Pavelka, N. Thieblemont, F. Willems, P. Ricciardi-Castagnoli, M. Goldman, M. Capron, et al. Double-stranded RNAs from the Helminth Parasite Schistosoma Activate TLR3 in Dendritic Cells J. Biol. Chem., January 7, 2005; 280(1): 277 - 283. [Abstract] [Full Text] [PDF]
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E. Latz, A. Visintin, T. Espevik, and D. T. Golenbock Mechanisms of TLR9 activation Innate Immunity, December 1, 2004; 10(6): 406 - 412. [Abstract] [PDF]
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M. Triantafilou, M. Manukyan, A. Mackie, S. Morath, T. Hartung, H. Heine, and K. Triantafilou Lipoteichoic Acid and Toll-like Receptor 2 Internalization and Targeting to the Golgi Are Lipid Raft-dependent J. Biol. Chem., September 24, 2004; 279(39): 40882 - 40889. [Abstract] [Full Text] [PDF]
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N. Nilsen, U. Nonstad, N. Khan, C. F. Knetter, S. Akira, A. Sundan, T. Espevik, and E. Lien Lipopolysaccharide and Double-stranded RNA Up-regulate Toll-like Receptor 2 Independently of Myeloid Differentiation Factor 88 J. Biol. Chem., September 17, 2004; 279(38): 39727 - 39735. [Abstract] [Full Text] [PDF]
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 M. Triantafilou, S. Morath, A. Mackie, T. Hartung, and K. Triantafilou Lateral diffusion of Toll-like receptors reveals that they are transiently confined within lipid rafts on the plasma membrane J. Cell Sci., September 1, 2004; 117(17): 4007 - 4014. [Abstract] [Full Text] [PDF]
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A. V. Bocharov, I. N. Baranova, T. G. Vishnyakova, A. T. Remaley, G. Csako, F. Thomas, A. P. Patterson, and T. L. Eggerman Targeting of Scavenger Receptor Class B Type I by Synthetic Amphipathic {alpha}-Helical-containing Peptides Blocks Lipopolysaccharide (LPS) Uptake and LPS-induced Pro-inflammatory Cytokine Responses in THP-1 Monocyte Cells J. Biol. Chem., August 20, 2004; 279(34): 36072 - 36082. [Abstract] [Full Text] [PDF]
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 S.-M. Lin, C. W. Frevert, O. Kajikawa, M. M. Wurfel, K. Ballman, S. Mongovin, V. A. Wong, A. Selk, and T. R. Martin Differential Regulation of Membrane CD14 Expression and Endotoxin-Tolerance in Alveolar Macrophages Am. J. Respir. Cell Mol. Biol., August 1, 2004; 31(2): 162 - 170. [Abstract] [Full Text] [PDF]
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 K. Funami, M. Matsumoto, H. Oshiumi, T. Akazawa, A. Yamamoto, and T. Seya The cytoplasmic 'linker region' in Toll-like receptor 3 controls receptor localization and signaling Int. Immunol., August 1, 2004; 16(8): 1143 - 1154. [Abstract] [Full Text] [PDF]
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S. Dunzendorfer, H.-K. Lee, K. Soldau, and P. S. Tobias TLR4 Is the Signaling but Not the Lipopolysaccharide Uptake Receptor J. Immunol., July 15, 2004; 173(2): 1166 - 1170. [Abstract] [Full Text] [PDF]
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C. A. Leifer, M. N. Kennedy, A. Mazzoni, C. Lee, M. J. Kruhlak, and D. M. Segal TLR9 Is Localized in the Endoplasmic Reticulum Prior to Stimulation J. Immunol., July 15, 2004; 173(2): 1179 - 1183. [Abstract] [Full Text] [PDF]
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A. Gruber, M. Mancek, H. Wagner, C. J. Kirschning, and R. Jerala Structural Model of MD-2 and Functional Role of Its Basic Amino Acid Clusters Involved in Cellular Lipopolysaccharide Recognition J. Biol. Chem., July 2, 2004; 279(27): 28475 - 28482. [Abstract] [Full Text] [PDF]
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 A. Vendeville, F. Rayne, A. Bonhoure, N. Bettache, P. Montcourrier, and B. Beaumelle HIV-1 Tat Enters T Cells Using Coated Pits before Translocating from Acidified Endosomes and Eliciting Biological Responses Mol. Biol. Cell, May 1, 2004; 15(5): 2347 - 2360. [Abstract] [Full Text] [PDF]
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T. L. Gioannini, A. Teghanemt, D. Zhang, N. P. Coussens, W. Dockstader, S. Ramaswamy, and J. P. Weiss Isolation of an endotoxin-MD-2 complex that produces Toll-like receptor 4-dependent cell activation at picomolar concentrations PNAS, March 23, 2004; 101(12): 4186 - 4191. [Abstract] [Full Text] [PDF]
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E. Latz, J. Franko, D. T. Golenbock, and J. R. Schreiber Haemophilus influenzae Type b-Outer Membrane Protein Complex Glycoconjugate Vaccine Induces Cytokine Production by Engaging Human Toll-Like Receptor 2 (TLR2) and Requires the Presence of TLR2 for Optimal Immunogenicity J. Immunol., February 15, 2004; 172(4): 2431 - 2438. [Abstract] [Full Text] [PDF]
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E. A. Kurt-Jones, M. Chan, S. Zhou, J. Wang, G. Reed, R. Bronson, M. M. Arnold, D. M. Knipe, and R. W. Finberg Herpes simplex virus 1 interaction with Toll-like receptor 2 contributes to lethal encephalitis PNAS, February 3, 2004; 101(5): 1315 - 1320. [Abstract] [Full Text] [PDF]
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L. Guillot, S. Medjane, K. Le-Barillec, V. Balloy, C. Danel, M. Chignard, and M. Si-Tahar Response of Human Pulmonary Epithelial Cells to Lipopolysaccharide Involves Toll-like Receptor 4 (TLR4)-dependent Signaling Pathways: EVIDENCE FOR AN INTRACELLULAR COMPARTMENTALIZATION OF TLR4 J. Biol. Chem., January 23, 2004; 279(4): 2712 - 2718. [Abstract] [Full Text] [PDF]
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J.-H. Lee, L. Del Sorbo, S. Uhlig, G. A. Porro, T. Whitehead, S. Voglis, M. Liu, A. S. Slutsky, and H. Zhang Intercellular Adhesion Molecule-1 Mediates Cellular Cross-Talk between Parenchymal and Immune Cells after Lipopolysaccharide Neutralization J. Immunol., January 1, 2004; 172(1): 608 - 616. [Abstract] [Full Text] [PDF]
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M. M. Monick, T. O. Yarovinsky, L. S. Powers, N. S. Butler, A. B. Carter, G. Gudmundsson, and G. W. Hunninghake Respiratory Syncytial Virus Up-regulates TLR4 and Sensitizes Airway Epithelial Cells to Endotoxin J. Biol. Chem., December 26, 2003; 278(52): 53035 - 53044. [Abstract] [Full Text] [PDF]
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H. Oshiumi, M. Sasai, K. Shida, T. Fujita, M. Matsumoto, and T. Seya TIR-containing Adapter Molecule (TICAM)-2, a Bridging Adapter Recruiting to Toll-like Receptor 4 TICAM-1 That Induces Interferon-{beta} J. Biol. Chem., December 12, 2003; 278(50): 49751 - 49762. [Abstract] [Full Text] [PDF]
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E. Latz, A. Visintin, E. Lien, K. A. Fitzgerald, T. Espevik, and D. T. Golenbock The LPS receptor generates inflammatory signals from the cell surface Innate Immunity, December 1, 2003; 9(6): 375 - 380. [Abstract] [PDF]
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A. Visintin, E. Latz, B. G. Monks, T. Espevik, and D. T. Golenbock Lysines 128 and 132 Enable Lipopolysaccharide Binding to MD-2, Leading to Toll-like Receptor-4 Aggregation and Signal Transduction J. Biol. Chem., November 28, 2003; 278(48): 48313 - 48320. [Abstract] [Full Text] [PDF]
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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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K. A. Fitzgerald, D. C. Rowe, B. J. Barnes, D. R. Caffrey, A. Visintin, E. Latz, B. Monks, P. M. Pitha, and D. T. Golenbock LPS-TLR4 Signaling to IRF-3/7 and NF-{kappa}B Involves the Toll Adapters TRAM and TRIF J. Exp. Med., October 6, 2003; 198(7): 1043 - 1055. [Abstract] [Full Text] [PDF]
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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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F. Sandor, E. Latz, F. Re, L. Mandell, G. Repik, D. T. Golenbock, T. Espevik, E. A. Kurt-Jones, and R. W. Finberg Importance of extra- and intracellular domains of TLR1 and TLR2 in NF{kappa}B signaling J. Cell Biol., September 15, 2003; 162(6): 1099 - 1110. [Abstract] [Full Text] [PDF]
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I. Sabroe, R. C. Read, M. K. B. Whyte, D. H. Dockrell, S. N. Vogel, and S. K. Dower Toll-Like Receptors in Health and Disease: Complex Questions Remain J. Immunol., August 15, 2003; 171(4): 1630 - 1635. [Full Text] [PDF]
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