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The Journal of Immunology, 2000, 165: 618-622.
Copyright © 2000 by The American Association of Immunologists


CUTTING EDGE

Cutting Edge: Repurification of Lipopolysaccharide Eliminates Signaling Through Both Human and Murine Toll-Like Receptor 21

Matthew Hirschfeld*, Ying Ma*, John H. Weis*, Stefanie N. Vogel{dagger} and Janis J. Weis2,*

* Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT 84132; and {dagger} Department of Microbiology and Immunology, Uniformed Services University of the Health Sciences, Bethesda, MD 20814

Toll-like receptor (TLR) 2 has recently been associated with cellular responses to numerous microbial products, including LPS and bacterial lipoproteins. However, many preparations of LPS contain low concentrations of highly bioactive contaminants described previously as "endotoxin protein," suggesting that these contaminants could be responsible for the TLR2-mediated signaling observed upon LPS stimulation. To test this hypothesis, commercial preparations of LPS were subjected to a modified phenol re-extraction protocol to eliminate endotoxin protein. While it did not influence the ability to stimulate cells from wild-type mice, repurification eliminated the ability of LPS to activate cells from C3H/HeJ (Lpsd) mice. Additionally, only cell lines transfected with human TLR4, but not human or murine TLR2, acquired responsiveness to both re-extracted LPS and to a protein-free, synthetic preparation of lipid A. These results suggest that neither human nor murine TLR2 plays a role in LPS signaling in the absence of contaminating endotoxin protein.




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A. Visintin, K. A. Halmen, N. Khan, B. G. Monks, D. T. Golenbock, and E. Lien
MD-2 expression is not required for cell surface targeting of Toll-like receptor 4 (TLR4)
J. Leukoc. Biol., December 1, 2006; 80(6): 1584 - 1592.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
D. J. Davidson, A. J. Currie, D. M. E. Bowdish, K. L. Brown, C. M. Rosenberger, R. C. Ma, J. Bylund, P. A. Campsall, A. Puel, C. Picard, et al.
IRAK-4 Mutation (Q293X): Rapid Detection and Characterization of Defective Post-Transcriptional TLR/IL-1R Responses in Human Myeloid and Non-Myeloid Cells
J. Immunol., December 1, 2006; 177(11): 8202 - 8211.
[Abstract] [Full Text] [PDF]


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Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
C. D. Coldren, J. A. Nick, K. R. Poch, M. D. Woolum, B. W. Fouty, J. M. O'Brien, M. P. Gruber, M. R. Zamora, D. Svetkauskaite, D. A. Richter, et al.
Functional and genomic changes induced by alveolar transmigration in human neutrophils
Am J Physiol Lung Cell Mol Physiol, December 1, 2006; 291(6): L1267 - L1276.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
K. W. Boehme, M. Guerrero, and T. Compton
Human Cytomegalovirus Envelope Glycoproteins B and H Are Necessary for TLR2 Activation in Permissive Cells
J. Immunol., November 15, 2006; 177(10): 7094 - 7102.
[Abstract] [Full Text] [PDF]


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Am. J. Physiol. Heart Circ. Physiol.Home page
X. Yang, V. Murthy, K. Schultz, J. B. Tatro, K. A. Fitzgerald, and D. Beasley
Toll-like receptor 3 signaling evokes a proinflammatory and proliferative phenotype in human vascular smooth muscle cells
Am J Physiol Heart Circ Physiol, November 1, 2006; 291(5): H2334 - H2343.
[Abstract] [Full Text] [PDF]


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CVIHome page
S. Ohta, U. Bahrun, R. Shimazu, H. Matsushita, K. Fukudome, and M. Kimoto
Induction of Long-Term Lipopolysaccharide Tolerance by an Agonistic Monoclonal Antibody to the Toll-Like Receptor 4/MD-2 Complex
Clin. Vaccine Immunol., October 1, 2006; 13(10): 1131 - 1136.
[Abstract] [Full Text] [PDF]


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J. Leukoc. Biol.Home page
H. Li, S. Nookala, X. R. Bina, J. E. Bina, and F. Re
Innate immune response to Francisella tularensis is mediated by TLR2 and caspase-1 activation
J. Leukoc. Biol., October 1, 2006; 80(4): 766 - 773.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
J. Rodo, L. A. Goncalves, J. Demengeot, A. Coutinho, and C. Penha-Goncalves
MHC Class II Molecules Control Murine B Cell Responsiveness to Lipopolysaccharide Stimulation
J. Immunol., October 1, 2006; 177(7): 4620 - 4626.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
A. W. Lee, L. Hertel, R. K. Louie, T. Burster, V. Lacaille, A. Pashine, D. A. Abate, E. S. Mocarski, and E. D. Mellins
Human Cytomegalovirus Alters Localization of MHC Class II and Dendrite Morphology in Mature Langerhans Cells
J. Immunol., September 15, 2006; 177(6): 3960 - 3971.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
M. Severa, E. M. Coccia, and K. A. Fitzgerald
Toll-like Receptor-dependent and -independent Viperin Gene Expression and Counter-regulation by PRDI-binding Factor-1/BLIMP1
J. Biol. Chem., September 8, 2006; 281(36): 26188 - 26195.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
J. M. Marques, R. J. Rodrigues, A. C. de Magalhaes-Sant'Ana, and T. Goncalves
Saccharomyces cerevisiae Hog1 Protein Phosphorylation upon Exposure to Bacterial Endotoxin
J. Biol. Chem., August 25, 2006; 281(34): 24687 - 24694.
[Abstract] [Full Text] [PDF]


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Infect. Immun.Home page
T. H. Harris, N. M. Cooney, J. M. Mansfield, and D. M. Paulnock
Signal transduction, gene transcription, and cytokine production triggered in macrophages by exposure to trypanosome DNA.
Infect. Immun., August 1, 2006; 74(8): 4530 - 4537.
[Abstract] [Full Text] [PDF]


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Innate ImmunityHome page
J. L. Shoenfelt and M. J. Fenton
TLR2- and TLR4-dependent activation of STAT1 serine phosphorylation in murine macrophages is protein kinase C-{delta}-independent
Innate Immunity, August 1, 2006; 12(4): 231 - 240.
[Abstract] [PDF]


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J. Leukoc. Biol.Home page
B. Gao, Y. Wang, and M.-F. Tsan
The heat sensitivity of cytokine-inducing effect of lipopolysaccharide
J. Leukoc. Biol., August 1, 2006; 80(2): 359 - 366.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
H. J. Brown, H. R. Lock, S. H. Sacks, and M. G. Robson
TLR2 Stimulation of Intrinsic Renal Cells in the Induction of Immune-Mediated Glomerulonephritis
J. Immunol., August 1, 2006; 177(3): 1925 - 1931.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
K. A. Scheibner, M. A. Lutz, S. Boodoo, M. J. Fenton, J. D. Powell, and M. R. Horton
Hyaluronan Fragments Act as an Endogenous Danger Signal by Engaging TLR2
J. Immunol., July 15, 2006; 177(2): 1272 - 1281.
[Abstract] [Full Text] [PDF]


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J. Leukoc. Biol.Home page
L. K. Weaver, K. A. Hintz-Goldstein, P. A. Pioli, K. Wardwell, N. Qureshi, S. N. Vogel, and P. M. Guyre
Pivotal Advance: Activation of cell surface Toll-like receptors causes shedding of the hemoglobin scavenger receptor CD163
J. Leukoc. Biol., July 1, 2006; 80(1): 26 - 35.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
C. N. Renn, D. J. Sanchez, M. T. Ochoa, A. J. Legaspi, C.-K. Oh, P. T. Liu, S. R. Krutzik, P. A. Sieling, G. Cheng, and R. L. Modlin
TLR Activation of Langerhans Cell-Like Dendritic Cells Triggers an Antiviral Immune Response
J. Immunol., July 1, 2006; 177(1): 298 - 305.
[Abstract] [Full Text] [PDF]


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J. Am. Soc. Nephrol.Home page
H. J. Brown, S. H. Sacks, and M. G. Robson
Toll-Like Receptor 2 Agonists Exacerbate Accelerated Nephrotoxic Nephritis
J. Am. Soc. Nephrol., July 1, 2006; 17(7): 1931 - 1939.
[Abstract] [Full Text] [PDF]


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Innate ImmunityHome page
A. E. Medvedev, I. Sabroe, J. D. Hasday, and S. N. Vogel
Invited review: Tolerance to microbial TLR ligands: molecular mechanisms and relevance to disease
Innate Immunity, June 1, 2006; 12(3): 133 - 150.
[Abstract] [PDF]


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Innate ImmunityHome page
O. Dehus, T. Hartung, and C. Hermann
Endotoxin evaluation of eleven lipopolysaccharides by whole blood assay does not always correlate with Limulus amebocyte lysate assay
Innate Immunity, June 1, 2006; 12(3): 171 - 180.
[Abstract] [PDF]


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J. Leukoc. Biol.Home page
G. Gatti, V. Rivero, R. D. Motrich, and M. Maccioni
Prostate epithelial cells can act as early sensors of infection by up-regulating TLR4 expression and proinflammatory mediators upon LPS stimulation
J. Leukoc. Biol., May 1, 2006; 79(5): 989 - 998.
[Abstract] [Full Text] [PDF]


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Proc. Natl. Acad. Sci. USAHome page
D. C. Rowe, A. F. McGettrick, E. Latz, B. G. Monks, N. J. Gay, M. Yamamoto, S. Akira, L. A. O'Neill, K. A. Fitzgerald, and D. T. Golenbock
The myristoylation of TRIF-related adaptor molecule is essential for Toll-like receptor 4 signal transduction
PNAS, April 18, 2006; 103(16): 6299 - 6304.
[Abstract] [Full Text] [PDF]


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Proc. Natl. Acad. Sci. USAHome page
K. Zacharowski, P. A. Zacharowski, A. Koch, A. Baban, N. Tran, R. Berkels, C. Papewalis, K. Schulze-Osthoff, P. Knuefermann, U. Zahringer, et al.
Toll-like receptor 4 plays a crucial role in the immune-adrenal response to systemic inflammatory response syndrome
PNAS, April 18, 2006; 103(16): 6392 - 6397.
[Abstract] [Full Text] [PDF]


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CVIHome page
H. S. Seo, J. H. Kim, and M. H. Nahm
Platelet-Activating Factor-Acetylhydrolase Can Monodeacylate and Inactivate Lipoteichoic Acid
Clin. Vaccine Immunol., April 1, 2006; 13(4): 452 - 458.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
L. R. Young, M. T. Borchers, H. L. Allen, R. S. Gibbons, and F. X. McCormack
Lung-Restricted Macrophage Activation in the Pearl Mouse Model of Hermansky-Pudlak Syndrome
J. Immunol., April 1, 2006; 176(7): 4361 - 4368.
[Abstract] [Full Text] [PDF]


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Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
J. Fan, Y. Li, Y. Vodovotz, T. R. Billiar, and M. A. Wilson
Hemorrhagic shock-activated neutrophils augment TLR4 signaling-induced TLR2 upregulation in alveolar macrophages: role in hemorrhage-primed lung inflammation
Am J Physiol Lung Cell Mol Physiol, April 1, 2006; 290(4): L738 - L746.
[Abstract] [Full Text] [PDF]


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Am. J. Physiol. Heart Circ. Physiol.Home page
R. E. Rumbaut, R. V. Bellera, J. K. Randhawa, C. N. Shrimpton, S. K. Dasgupta, J.-F. Dong, and A. R. Burns
Endotoxin enhances microvascular thrombosis in mouse cremaster venules via a TLR4-dependent, neutrophil-independent mechanism
Am J Physiol Heart Circ Physiol, April 1, 2006; 290(4): H1671 - H1679.
[Abstract] [Full Text] [PDF]


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Am. J. Physiol. Cell Physiol.Home page
J. S. Park, F. Gamboni-Robertson, Q. He, D. Svetkauskaite, J.-Y. Kim, D. Strassheim, J.-W. Sohn, S. Yamada, I. Maruyama, A. Banerjee, et al.
High mobility group box 1 protein interacts with multiple Toll-like receptors
Am J Physiol Cell Physiol, March 1, 2006; 290(3): C917 - C924.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
J. Bylund, L.-A. Burgess, P. Cescutti, R. K. Ernst, and D. P. Speert
Exopolysaccharides from Burkholderia cenocepacia Inhibit Neutrophil Chemotaxis and Scavenge Reactive Oxygen Species
J. Biol. Chem., February 3, 2006; 281(5): 2526 - 2532.
[Abstract] [Full Text] [PDF]


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Int ImmunolHome page
M. Hashimoto, K. Tawaratsumida, H. Kariya, K. Aoyama, T. Tamura, and Y. Suda
Lipoprotein is a predominant Toll-like receptor 2 ligand in Staphylococcus aureus cell wall components
Int. Immunol., February 1, 2006; 18(2): 355 - 362.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
T. B. Thornley, M. A. Brehm, T. G. Markees, L. D. Shultz, J. P. Mordes, R. M. Welsh, A. A. Rossini, and D. L. Greiner
TLR Agonists Abrogate Costimulation Blockade-Induced Prolongation of Skin Allografts
J. Immunol., February 1, 2006; 176(3): 1561 - 1570.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
C. M. O'Connell, I. A. Ionova, A. J. Quayle, A. Visintin, and R. R. Ingalls
Localization of TLR2 and MyD88 to Chlamydia trachomatis Inclusions: EVIDENCE FOR SIGNALING BY INTRACELLULAR TLR2 DURING INFECTION WITH AN OBLIGATE INTRACELLULAR PATHOGEN
J. Biol. Chem., January 20, 2006; 281(3): 1652 - 1659.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
S. H. Han, J. H. Kim, H. S. Seo, M. H. Martin, G.-H. Chung, S. M. Michalek, and M. H. Nahm
Lipoteichoic Acid-Induced Nitric Oxide Production Depends on the Activation of Platelet-Activating Factor Receptor and Jak2
J. Immunol., January 1, 2006; 176(1): 573 - 579.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
K. L. Williams, J. D. Lich, J. A. Duncan, W. Reed, P. Rallabhandi, C. Moore, S. Kurtz, V. M. Coffield, M. A. Accavitti-Loper, L. Su, et al.
The CATERPILLER Protein Monarch-1 Is an Antagonist of Toll-like Receptor-, Tumor Necrosis Factor {alpha}-, and Mycobacterium tuberculosis-induced Pro-inflammatory Signals
J. Biol. Chem., December 2, 2005; 280(48): 39914 - 39924.
[Abstract] [Full Text] [PDF]


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Innate ImmunityHome page
S. Morath, S. von Aulock, and T. Hartung
Structure/function relationships of lipoteichoic acids
Innate Immunity, December 1, 2005; 11(6): 348 - 356.
[Abstract] [PDF]


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J. Leukoc. Biol.Home page
L. Romics Jr, A. Dolganiuc, A. Velayudham, K. Kodys, P. Mandrekar, D. Golenbock, E. Kurt-Jones, and G. Szabo
Toll-like receptor 2 mediates inflammatory cytokine induction but not sensitization for liver injury by Propioni- bacterium acnes
J. Leukoc. Biol., December 1, 2005; 78(6): 1255 - 1264.
[Abstract] [Full Text] [PDF]


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J. Leukoc. Biol.Home page
L. C. Parker, E. C. Jones, L. R. Prince, S. K. Dower, M. K. B. Whyte, and I. Sabroe
Endotoxin tolerance induces selective alterations in neutrophil function
J. Leukoc. Biol., December 1, 2005; 78(6): 1301 - 1305.
[Abstract] [Full Text] [PDF]




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