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(Journal of Leukocyte Biology. 2001;69:1036-1044.)
© 2001 by Society for Leukocyte Biology

Different Toll-like receptor agonists induce distinct macrophage responses

Bryan W. Jones*, Terry K. Means*, Kurt A. Heldwein*, Marc A. Keen{dagger}, Preston J. Hill{dagger}, John T. Belisle{dagger} and Matthew J. Fenton*

* The Pulmonary Center, Boston University School of Medicine, Boston Massachusetts, and
{dagger} Mycobacteria Research Laboratories, Department of Microbiology, Colorado State University, Ft. Collins, Colorado

Correspondence: Dr. Matthew J. Fenton, Pulmonary Center R-220, Boston University School of Medicine, 80 East Concord St., Boston MA 02118-2394. E-mail: mfenton{at}bu.edu

We previously reported that gram-negative bacterial lipopolysaccharide (LPS) activates cells via Toll-like receptor (TLR) 4, whereas the mycobacterial cell wall glycolipid lipoarabinomannan (LAM) activates cells via TLR2. We also identified a secreted TLR2 agonist activity in short-term culture filtrates of Mycobacterium tuberculosis bacilli, termed soluble tuberculosis factor (STF). Here we show that STF contains mannosylated phosphatidylinositol (PIM) and that purified PIM possesses TLR2 agonist activity. Stimulation of RAW 264.7 macrophages by LPS, LAM, STF, and PIM rapidly activated nuclear factor (NF)-{kappa}B, activator protein-1 (AP-1), and mitogen-activated protein (MAP) kinases. These TLR agonists induced similar levels of NF-{kappa}B and AP-1 DNA-binding activity, as well as trans-activation function. Unexpectedly, these TLR agonists induced tumor necrosis factor {alpha} secretion, whereas only LPS was capable of inducing interleukin-1ß and nitric oxide secretion. Thus, different TLR proteins are still capable of activating distinct cellular responses, in spite of their shared capacities to activate NF-{kappa}B, AP-1, and MAP kinases.

Key Words: signal transduction • tuberculosis • innate immunity


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A. Bafica, C. A. Scanga, M. L. Schito, S. Hieny, and A. Sher
Cutting Edge: In Vivo Induction of Integrated HIV-1 Expression by Mycobacteria Is Critically Dependent on Toll-Like Receptor 2
J. Immunol., August 1, 2003; 171(3): 1123 - 1127.
[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
S. B. Mizel, A. N. Honko, M. A. Moors, P. S. Smith, and A. P. West
Induction of Macrophage Nitric Oxide Production by Gram-Negative Flagellin Involves Signaling Via Heteromeric Toll-Like Receptor 5/Toll-Like Receptor 4 Complexes
J. Immunol., June 15, 2003; 170(12): 6217 - 6223.
[Abstract] [Full Text] [PDF]


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Innate ImmunityHome page
V. Toshchakov, B. W. Jones, A. Lentschat, A. Silva, P.-Y. Perera, K. Thomas, M. J. Cody, Shuling Zhang, B. R.G. Williams, J. Major, et al.
TLR2 and TLR4 agonists stimulate unique repertoires of host resistance genes in murine macrophages: interferon-{beta}-dependent signaling in TLR4-mediated responses
Innate Immunity, June 1, 2003; 9(3): 169 - 175.
[Abstract] [PDF]


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Proc. Natl. Acad. Sci. USAHome page
J. Lee, T.-H. Chuang, V. Redecke, L. She, P. M. Pitha, D. A. Carson, E. Raz, and H. B. Cottam
Molecular basis for the immunostimulatory activity of guanine nucleoside analogs: Activation of Toll-like receptor 7
PNAS, May 27, 2003; 100(11): 6646 - 6651.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
O. Equils, M. L. Schito, H. Karahashi, Z. Madak, A. Yarali, K. S. Michelsen, A. Sher, and M. Arditi
Toll-Like Receptor 2 (TLR2) and TLR9 Signaling Results in HIV-Long Terminal Repeat Trans-Activation and HIV Replication in HIV-1 Transgenic Mouse Spleen Cells: Implications of Simultaneous Activation of TLRs on HIV Replication
J. Immunol., May 15, 2003; 170(10): 5159 - 5164.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
P.-Y. Bochud, T. R. Hawn, and A. Aderem
Cutting Edge: A Toll-Like Receptor 2 Polymorphism That Is Associated with Lepromatous Leprosy Is Unable to Mediate Mycobacterial Signaling
J. Immunol., April 1, 2003; 170(7): 3451 - 3454.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
J. D. McCurdy, T. J. Olynych, L. H. Maher, and J. S. Marshall
Cutting Edge: Distinct Toll-Like Receptor 2 Activators Selectively Induce Different Classes of Mediator Production from Human Mast Cells
J. Immunol., February 15, 2003; 170(4): 1625 - 1629.
[Abstract] [Full Text] [PDF]


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Infect. Immun.Home page
M. L. Hart, D. A. Mosier, and S. K. Chapes
Toll-Like Receptor 4-Positive Macrophages Protect Mice from Pasteurella pneumotropica-Induced Pneumonia
Infect. Immun., February 1, 2003; 71(2): 663 - 670.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
G. Melmed, L. S. Thomas, N. Lee, S. Y. Tesfay, K. Lukasek, K. S. Michelsen, Y. Zhou, B. Hu, M. Arditi, and M. T. Abreu
Human Intestinal Epithelial Cells Are Broadly Unresponsive to Toll-Like Receptor 2-Dependent Bacterial Ligands: Implications for Host-Microbial Interactions in the Gut
J. Immunol., February 1, 2003; 170(3): 1406 - 1415.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
M. A. Dobrovolskaia, A. E. Medvedev, K. E. Thomas, N. Cuesta, V. Toshchakov, T. Ren, M. J. Cody, S. M. Michalek, N. R. Rice, and S. N. Vogel
Induction of In Vitro Reprogramming by Toll-Like Receptor (TLR)2 and TLR4 Agonists in Murine Macrophages: Effects of TLR "Homotolerance" Versus "Heterotolerance" on NF-{kappa}B Signaling Pathway Components
J. Immunol., January 1, 2003; 170(1): 508 - 519.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
E. K. Brint, K. A. Fitzgerald, P. Smith, A. J. Coyle, J.-C. Gutierrez-Ramos, P. G. Fallon, and L. A. J. O'Neill
Characterization of Signaling Pathways Activated by the Interleukin 1 (IL-1) Receptor Homologue T1/ST2. A ROLE FOR JUN N-TERMINAL KINASE IN IL-4 INDUCTION
J. Biol. Chem., December 13, 2002; 277(51): 49205 - 49211.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
D. Schilling, K. Thomas, K. Nixdorff, S. N. Vogel, and M. J. Fenton
Toll-Like Receptor 4 and Toll-IL-1 Receptor Domain-Containing Adapter Protein (TIRAP)/Myeloid Differentiation Protein 88 Adapter-Like (Mal) Contribute to Maximal IL-6 Expression in Macrophages
J. Immunol., November 15, 2002; 169(10): 5874 - 5880.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
M. R. Horton, S. Boodoo, and J. D. Powell
NF-kappa B Activation Mediates the Cross-talk between Extracellular Matrix and Interferon-gamma (IFN-gamma ) Leading to Enhanced Monokine Induced by IFN-gamma (MIG) Expression in Macrophages
J. Biol. Chem., November 8, 2002; 277(46): 43757 - 43762.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
A. E. Medvedev, A. Lentschat, L. M. Wahl, D. T. Golenbock, and S. N. Vogel
Dysregulation of LPS-Induced Toll-Like Receptor 4-MyD88 Complex Formation and IL-1 Receptor-Associated Kinase 1 Activation in Endotoxin-Tolerant Cells
J. Immunol., November 1, 2002; 169(9): 5209 - 5216.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
A. D. Edwards, S. P. Manickasingham, R. Sporri, S. S. Diebold, O. Schulz, A. Sher, T. Kaisho, S. Akira, and C. Reis e Sousa
Microbial Recognition Via Toll-Like Receptor-Dependent and -Independent Pathways Determines the Cytokine Response of Murine Dendritic Cell Subsets to CD40 Triggering
J. Immunol., October 1, 2002; 169(7): 3652 - 3660.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
B. Abel, N. Thieblemont, V. J. F. Quesniaux, N. Brown, J. Mpagi, K. Miyake, F. Bihl, and B. Ryffel
Toll-Like Receptor 4 Expression Is Required to Control Chronic Mycobacterium tuberculosis Infection in Mice
J. Immunol., September 15, 2002; 169(6): 3155 - 3162.
[Abstract] [Full Text] [PDF]


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Infect. Immun.Home page
A. Blumenthal, S. Ehlers, M. Ernst, H.-D. Flad, and N. Reiling
Control of Mycobacterial Replication in Human Macrophages: Roles of Extracellular Signal-Regulated Kinases 1 and 2 and p38 Mitogen-Activated Protein Kinase Pathways
Infect. Immun., September 1, 2002; 70(9): 4961 - 4967.
[Abstract] [Full Text] [PDF]


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Int ImmunolHome page
S. E. Applequist, R. P. A. Wallin, and H.-G. Ljunggren
Variable expression of Toll-like receptor in murine innate and adaptive immune cell lines
Int. Immunol., September 1, 2002; 14(9): 1065 - 1074.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
M. Guha and N. Mackman
The Phosphatidylinositol 3-Kinase-Akt Pathway Limits Lipopolysaccharide Activation of Signaling Pathways and Expression of Inflammatory Mediators in Human Monocytic Cells
J. Biol. Chem., August 23, 2002; 277(35): 32124 - 32132.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
T. Wang, W. P. Lafuse, K. Takeda, S. Akira, and B. S. Zwilling
Rapid Chromatin Remodeling of Toll-Like Receptor 2 Promoter During Infection of Macrophages with Mycobacterium avium
J. Immunol., July 15, 2002; 169(2): 795 - 801.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
V. Haehnel, L. Schwarzfischer, M. J. Fenton, and M. Rehli
Transcriptional Regulation of the Human Toll-Like Receptor 2 Gene in Monocytes and Macrophages
J. Immunol., June 1, 2002; 168(11): 5629 - 5637.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
V. S. Carl, K. Brown-Steinke, M. J. H. Nicklin, and M. F. Smith Jr.
Toll-like Receptor 2 and 4 (TLR2 and TLR4) Agonists Differentially Regulate Secretory Interleukin-1 Receptor Antagonist Gene Expression in Macrophages
J. Biol. Chem., May 10, 2002; 277(20): 17448 - 17456.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
B. G. Yipp, G. Andonegui, C. J. Howlett, S. M. Robbins, T. Hartung, M. Ho, and P. Kubes
Profound Differences in Leukocyte-Endothelial Cell Responses to Lipopolysaccharide Versus Lipoteichoic Acid
J. Immunol., May 1, 2002; 168(9): 4650 - 4658.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
M. Martin, J. Katz, S. N. Vogel, and S. M. Michalek
Differential Induction of Endotoxin Tolerance by Lipopolysaccharides Derived from Porphyromonas gingivalis and Escherichia coli
J. Immunol., November 1, 2001; 167(9): 5278 - 5285.
[Abstract] [Full Text] [PDF]


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Ann Rheum DisHome page
B W Jones, K A Heldwein, T K Means, J J Saukkonen, and M J Fenton
Differential roles of Toll-like receptors in the elicitation of proinflammatory responses by macrophages
Ann Rheum Dis, November 1, 2001; 60(90003): iii6 - 12.
[Abstract] [Full Text] [PDF]


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Proc. Natl. Acad. Sci. USAHome page
E. Trogan, R. P. Choudhury, H. M. Dansky, J. X. Rong, J. L. Breslow, and E. A. Fisher
Laser capture microdissection analysis of gene expression in macrophages from atherosclerotic lesions of apolipoprotein E-deficient mice
PNAS, February 19, 2002; 99(4): 2234 - 2239.
[Abstract] [Full Text] [PDF]