MyD88 Primes Macrophages for Full-Scale Activation by Interferon-γ yet Mediates Few Responses to Mycobacterium tuberculosis

Macrophages are activated from a resting state by a combination of cytokines and microbial products. Microbes are often sensed through Toll-like receptors signaling through MyD88. We used large-scale microarrays in multiple replicate experiments followed by stringent statistical analysis to compare gene expression in wild-type (WT) and MyD88−/− macrophages. We confirmed key results by quantitative reverse transcription polymerase chain reaction, Western blot, and enzyme-linked immunosorbent assay. Surprisingly, many genes, such as inducible nitric oxide synthase, IRG-1, IP-10, MIG, RANTES, and interleukin 6 were induced by interferon (IFN)-γ from 5- to 100-fold less extensively in MyD88−/− macrophages than in WT macrophages. Thus, widespread, full-scale activation of macrophages by IFN-γ requires MyD88. Analysis of the mechanism revealed that MyD88 mediates a process of self-priming by which resting macrophages produce a low level of tumor necrosis factor. This and other factors lead to basal activation of nuclear factor κB, which synergizes with IFN-γ for gene induction. In contrast, infection by live, virulent Mycobacterium tuberculosis (Mtb) activated macrophages largely through MyD88-independent pathways, and macrophages did not need MyD88 to kill Mtb in vitro. Thus, MyD88 plays a dynamic role in resting macrophages that supports IFN-γ–dependent activation, whereas macrophages can respond to a complex microbial stimulus, the tubercle bacillus, chiefly by other routes.

[1]  T. van der Poll,et al.  CD44 is a macrophage binding site for Mycobacterium tuberculosis that mediates macrophage recruitment and protective immunity against tuberculosis. , 2003, The Journal of clinical investigation.

[2]  C. Sousa Faculty Opinions recommendation of TICAM-1, an adaptor molecule that participates in Toll-like receptor 3-mediated interferon-beta induction. , 2003 .

[3]  C. Figdor,et al.  Expression of the dendritic cell-associated C-type lectin DC-SIGN by inflammatory matrix metalloproteinase-producing macrophages in rheumatoid arthritis synovium and interaction with intercellular adhesion molecule 3-positive T cells. , 2003, Arthritis and rheumatism.

[4]  D. Mosser,et al.  The many faces of macrophage activation , 2003, Journal of leukocyte biology.

[5]  J. Witztum,et al.  Minimally Modified LDL Binds to CD14, Induces Macrophage Spreading via TLR4/MD-2, and Inhibits Phagocytosis of Apoptotic Cells* , 2003, The Journal of Biological Chemistry.

[6]  O. Schwartz,et al.  DC-SIGN Is the Major Mycobacterium tuberculosis Receptor on Human Dendritic Cells , 2003, The Journal of experimental medicine.

[7]  T. Geijtenbeek,et al.  Mycobacteria Target DC-SIGN to Suppress Dendritic Cell Function , 2003, The Journal of experimental medicine.

[8]  S. Gordon Alternative activation of macrophages , 2003, Nature Reviews Immunology.

[9]  R. Flavell,et al.  The adaptor molecule TIRAP provides signalling specificity for Toll-like receptors , 2002, Nature.

[10]  S. Akira,et al.  Essential role for TIRAP in activation of the signalling cascade shared by TLR2 and TLR4 , 2002, Nature.

[11]  L. Kwak,et al.  Toll-Like Receptor 4-Dependent Activation of Dendritic Cells by β-Defensin 2 , 2002, Science.

[12]  E. Unanue,et al.  MyD88-Dependent but Toll-Like Receptor 2-Independent Innate Immunity to Listeria: No Role for Either in Macrophage Listericidal Activity1 , 2002, The Journal of Immunology.

[13]  S. Akira,et al.  Critical Roles of Myeloid Differentiation Factor 88-Dependent Proinflammatory Cytokine Release in Early Phase Clearance of Listeria monocytogenes in Mice1 , 2002, The Journal of Immunology.

[14]  R. Vabulas,et al.  Cutting Edge: Myeloid Differentiation Factor 88 Deficiency Improves Resistance Against Sepsis Caused by Polymicrobial Infection1 , 2002, The Journal of Immunology.

[15]  R. Modlin,et al.  Control of Mycobacterium tuberculosis through mammalian Toll-like receptors. , 2002, Current opinion in immunology.

[16]  M. Fenton,et al.  The role of Toll-like receptors in immunity against mycobacterial infection. , 2002, Microbes and infection.

[17]  A. Sher,et al.  Cutting Edge: MyD88 Is Required for Resistance to Toxoplasma gondii Infection and Regulates Parasite-Induced IL-12 Production by Dendritic Cells1 , 2002, The Journal of Immunology.

[18]  J. Roman,et al.  M. tuberculosis induction of matrix metalloproteinase-9: the role of mannose and receptor-mediated mechanisms. , 2002, American journal of physiology. Lung cellular and molecular physiology.

[19]  H. Schild,et al.  Heat shock proteins as ligands of toll-like receptors. , 2002, Current topics in microbiology and immunology.

[20]  L. Kwak,et al.  Toll-like receptor 4-dependent activation of dendritic cells by beta-defensin 2. , 2002, Science.

[21]  S. Akira,et al.  Lipopolysaccharide Stimulates the MyD88-Independent Pathway and Results in Activation of IFN-Regulatory Factor 3 and the Expression of a Subset of Lipopolysaccharide-Inducible Genes1 , 2001, The Journal of Immunology.

[22]  M. Fenton,et al.  Differential roles of Toll-like receptors in the elicitation of proinflammatory responses by macrophages , 2001, Annals of the rheumatic diseases.

[23]  T. Gingeras,et al.  Reprogramming of the Macrophage Transcriptome in Response to Interferon-γ and Mycobacterium tuberculosis , 2001, The Journal of experimental medicine.

[24]  S. Akira,et al.  Toll-like receptors: critical proteins linking innate and acquired immunity , 2001, Nature Immunology.

[25]  Amer A. Beg,et al.  An Essential Role of the NF-κB/Toll-Like Receptor Pathway in Induction of Inflammatory and Tissue-Repair Gene Expression by Necrotic Cells1 , 2001, The Journal of Immunology.

[26]  J. Keane,et al.  Differential Effects of a Toll-Like Receptor Antagonist on Mycobacterium tuberculosis-Induced Macrophage Responses1 , 2001, The Journal of Immunology.

[27]  J. Bijlsma Diagnosis and nonsurgical management of osteoarthritis , 2001, Annals of the rheumatic diseases.

[28]  T. Gingeras,et al.  Reprogramming of the macrophage transcriptome in response to interferon-gamma and Mycobacterium tuberculosis: signaling roles of nitric oxide synthase-2 and phagocyte oxidase. , 2001, The Journal of experimental medicine.

[29]  S. Akira,et al.  Cutting Edge: TLR2-Deficient and MyD88-Deficient Mice Are Highly Susceptible to Staphylococcus aureus Infection1 , 2000, The Journal of Immunology.

[30]  P. Srivastava,et al.  Necrotic but not apoptotic cell death releases heat shock proteins, which deliver a partial maturation signal to dendritic cells and activate the NF-kappa B pathway. , 2000, International immunology.

[31]  E. Cesarman,et al.  Inhibition of NF-kappaB induces apoptosis of KSHV-infected primary effusion lymphoma cells. , 2000, Blood.

[32]  T. van der Poll,et al.  Interleukin-1 signaling is essential for host defense during murine pulmonary tuberculosis. , 2000, The Journal of infectious diseases.

[33]  I. Orme,et al.  Interleukin-6 Induces Early Gamma Interferon Production in the Infected Lung but Is Not Required for Generation of Specific Immunity to Mycobacterium tuberculosisInfection , 2000, Infection and Immunity.

[34]  Y. Iwakura,et al.  Protective Role of Interleukin-1 in Mycobacterial Infection in IL-1 α/β Double-Knockout Mice , 2000, Laboratory Investigation.

[35]  S. Teitelbaum,et al.  Tumor necrosis factor alpha regulates alpha(v)beta5 integrin expression by osteoclast precursors in vitro and in vivo. , 2000, Endocrinology.

[36]  Y. Iwakura,et al.  Protective role of interleukin-1 in mycobacterial infection in IL-1 alpha/beta double-knockout mice. , 2000, Laboratory investigation; a journal of technical methods and pathology.

[37]  Shi Wei,et al.  Tumor Necrosis Factor α Regulatesα vβ5 Integrin Expression by Osteoclast Precursors in Vitro and in Vivo1. , 2000, Endocrinology.

[38]  D. Golenbock,et al.  Human toll-like receptors mediate cellular activation by Mycobacterium tuberculosis. , 1999, Journal of immunology.

[39]  S. Akira,et al.  Unresponsiveness of MyD88-deficient mice to endotoxin. , 1999, Immunity.

[40]  C. Janeway,et al.  MyD88 is an adaptor protein in the hToll/IL-1 receptor family signaling pathways. , 1998, Molecular cell.

[41]  S. Akira,et al.  Targeted disruption of the MyD88 gene results in loss of IL-1- and IL-18-mediated function. , 1998, Immunity.

[42]  J. Ernst,et al.  Macrophage Receptors for Mycobacterium tuberculosis , 1998, Infection and Immunity.

[43]  S. Kaufmann,et al.  Copyright © 1997, American Society for Microbiology Lethal Tuberculosis in Interleukin-6-Deficient Mutant Mice , 1997 .

[44]  M. Wainberg,et al.  Induction of relA(p65) and I kappa B alpha subunit expression during differentiation of human peripheral blood monocytes to macrophages. , 1997, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[45]  T. Miyata,et al.  The receptor for advanced glycation end products (RAGE) is a central mediator of the interaction of AGE-beta2microglobulin with human mononuclear phagocytes via an oxidant-sensitive pathway. Implications for the pathogenesis of dialysis-related amyloidosis. , 1996, The Journal of clinical investigation.

[46]  A. Tomasz,et al.  CD14 is a pattern recognition receptor. , 1994, Immunity.

[47]  W. Fanslow,et al.  CD40 expression by human monocytes: regulation by cytokines and activation of monocytes by the ligand for CD40 , 1993, The Journal of experimental medicine.

[48]  T. Lee,et al.  Mechanisms of stimulation of interleukin-1 beta and tumor necrosis factor-alpha by Mycobacterium tuberculosis components. , 1993, The Journal of clinical investigation.

[49]  Terry D. Lee,et al.  Cloning and characterization of inducible nitric oxide synthase from mouse macrophages. , 1992, Science.

[50]  M. Fresno,et al.  Synergism between tumor necrosis factor‐α and interferon‐γ on macrophage activation for the killing of intracellular Trypanosoma cruzi through a nitric oxide‐dependent mechanism , 1992 .

[51]  M. Fresno,et al.  Synergism between tumor necrosis factor-alpha and interferon-gamma on macrophage activation for the killing of intracellular Trypanosoma cruzi through a nitric oxide-dependent mechanism. , 1992, European journal of immunology.

[52]  D. Riches,et al.  Expression of interferon-beta during the triggering phase of macrophage cytocidal activation. Evidence for an autocrine/paracrine role in the regulation of this state. , 1991, The Journal of biological chemistry.

[53]  J. Sypek,et al.  Antileishmanial defense in macrophages triggered by tumor necrosis factor expressed on CD4+ T lymphocyte plasma membrane , 1991, The Journal of experimental medicine.

[54]  P. Bailly,et al.  Molecular characterization of the human macrophage mannose receptor: demonstration of multiple carbohydrate recognition-like domains and phagocytosis of yeasts in Cos-1 cells , 1990, The Journal of experimental medicine.

[55]  C. Nathan,et al.  Release of reactive nitrogen intermediates and reactive oxygen intermediates from mouse peritoneal macrophages. Comparison of activating cytokines and evidence for independent production. , 1988, Journal of immunology.

[56]  S. Haskill,et al.  Adherence induces selective mRNA expression of monocyte mediators and proto-oncogenes. , 1988, Journal of immunology.

[57]  E. Unanue,et al.  Regulation of interleukin 1 gene expression by adherence and lipopolysaccharide. , 1987, Journal of immunology.

[58]  R. Mertelsmann,et al.  Activation of human macrophages. Comparison of other cytokines with interferon-gamma , 1984, Journal of Experimental Medicine.

[59]  T. Hamilton,et al.  The cell biology of macrophage activation. , 1984, Annual review of immunology.

[60]  M. Meltzer Macrophage activation for tumor cytotoxicity: characterization of priming and trigger signals during lymphokine activation. , 1981, Journal of immunology.

[61]  S. Russell,et al.  Activation of mouse macrophages for tumor cell killing. I. Quantitative analysis of interactions between lymphokine and lipopolysaccharide. , 1981, Journal of immunology.

[62]  H. Chapman,et al.  Modulation of the tumoricidal function of activated macrophages by bacterial endotoxin and mammalian macrophage activation factor (s). , 1979, Advances in experimental medicine and biology.

[63]  C. Nathan,et al.  ALTERATIONS OF MACROPHAGE FUNCTIONS BY MEDIATORS FROM LYMPHOCYTES , 1971, The Journal of experimental medicine.

[64]  G. Mackaness THE INFLUENCE OF IMMUNOLOGICALLY COMMITTED LYMPHOID CELLS ON MACROPHAGE ACTIVITY IN VIVO , 1969, The Journal of experimental medicine.