Abstract
The innate immunity is an essential step as the front line of host defense, and its aberrant activation particularly in response to nucleic acids is closely related to the pathogenesis of autoimmune and inflammatory diseases. Characterization of the innate immune signalings may provide a pathophysiological insight for better understanding of human diseases. Nucleic acid-mediated activation of pattern recognition receptors triggers the activation of two major intracellular signaling pathways, which are dependent on NF-κB and interferon regulatory factors, transcriptional factors. This leads to the subsequent induction of inflammatory cytokines and type I and III interferons. In this chapter, we first overview the representative families of nucleic acid sensors and their ligands and then show the fundamental techniques for extracellular or intracellular stimulation with these nucleic acid ligands and for detection of innate immune response, that is, IFN and proinflammatory cytokine induction, as assessed by luciferase assay, quantitative RT-PCR (qRT-PCR), and enzyme-linked immunosorbent assay.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Takaoka A, Yanai H (2006) Interferon signalling network in innate defence. Cell Microbiol 8:907–922
Mogensen TH (2009) Pathogen recognition and inflammatory signaling in innate immune defenses. Clin Microbiol Rev 22:240–273
Takeuchi O, Akira S (2010) Pattern recognition receptors and inflammation. Cell 140: 805–820
Vladimer GI, Marty-Roix R, Ghosh S, Weng D, Lien E (2013) Inflammasomes and host defenses against bacterial infections. Curr Opin Microbiol 16:23–31
Kono H, Rock KL (2008) How dying cells alert the immune system to danger. Nat Cell Biol 8:279–289
Jounai N, Kobiyama K, Takeshita F, Ishii KJ (2012) Recognition of damage-associated molecular patterns related to nucleic acids during inflammation and vaccination. Front Cell Infect Microbiol 2:168. doi:10.3389/fcimb.2012.00168
Barrat FJ, Meeker T, Gregorio J, Chan JH, Uematsu S, Akira S et al (2005) Nucleic acids of mammalian origin can act as endogenous ligands for Toll-like receptors and may promote systemic lupus erythematosus. J Exp Med 202:1131–1139
Barbalat R, Ewald SE, Mouchess ML, Barton GM (2011) Nucleic acid recognition by the innate immune system. Annu Rev Immunol 29:185–214
Takaoka A, Taniguchi T (2008) Cytosolic DNA recognition for triggering innate immune responses. Adv Drug Deliv Rev 60: 847–857
Oldenburg M, Krüger A, Ferstl R, Kaufmann A, Nees G, Sigmund A et al (2012) TLR13 recognizes bacterial 23S rRNA devoid of erythromycin resistance-forming modification. Science 337:1111–1115
Hemmi H, Takeuchi O, Kawai T, Kaisho T, Sato S, Sanjo H et al (2000) A Toll-like receptor recognizes bacterial DNA. Nature 408:740–745
Onoguchi K, Yoneyama M, Fujita T (2011) Retinoic acid-inducible gene-I-like receptors. J Interferon Cytokine Res 31:27–31
Yoneyama M, Fujita T (2009) RNA recognition and signal transduction by RIG-I-like receptors. Immunol Rev 227:54–65
Rehwinkel J, Reis e Sousa C (2010) RIGorous detection: exposing virus through RNA sensing. Science 327:284–286
Yoneyama M, Kikuchi M, Natsukawa T, Shinobu N, Imaizumi T, Miyagishi M et al (2004) The RNA helicase RIG-I has an essential function in double-stranded RNA-induced innate antiviral responses. Nat Immunol 5:730–737
Fujita T (2009) A nonself RNA pattern: tri-p to panhandle. Immunity 31:4–5
Takeuchi O, Akira S (2009) Innate immunity to virus infection. Immunol Rev 227:75–86
Kato H, Takeuchi O, Mikamo-Satoh E, Hirai R, Kawai T, Matsushita K et al (2008) Length-dependent recognition of double-stranded ribonucleic acids by retinoic acid-inducible gene-I and melanoma differentiation-associated gene 5. J Exp Med 205:1601–1610
Ablasser A, Bauernfeind F, Hartmann G, Latz E, Fitzgerald KA, Hornung V (2009) RIG-I-dependent sensing of poly(dA:dT) through the induction of an RNA polymerase III-transcribed RNA intermediate. Nat Immunol 10:1065–1072
Chiu Y-H, MacMillan JB, Chen ZJ (2009) RNA polymerase III detects cytosolic DNA and induces type I interferons through the RIG-I pathway. Cell 138:576–591
Sabbah A, Chang TH, Harnack R, Frohlich V, Tominaga K, Dube PH et al (2009) Activation of innate immune antiviral responses by Nod2. Nat Immunol 10:1073–1080
Takaoka A, Wang Z, Choi MK, Yanai H, Negishi H, Ban T et al (2007) DAI (DLM-1/ZBP1) is a cytosolic DNA sensor and an activator of innate immune response. Nature 448:501–505
Unterholzner L, Keating SE, Baran M, Horan KA, Jensen SB, Sharma S et al (2010) IFI16 is an innate immune sensor for intracellular DNA. Nat Immunol 11:997–1004
Zhang Z, Yuan B, Bao M, Lu N, Kim T, Liu Y-J (2011) The helicase DDX41 senses intracellular DNA mediated by the adaptor STING in dendritic cells. Nat Immunol 12:959–965
Stetson DB, Medzhitov R (2006) Recognition of cytosolic DNA activates an IRF3-dependent innate immune response. Immunity 24:93–103
Burdette DL, Monroe KM, Sotelo-Troha K, Iwig JS, Eckert B, Hyodo M et al (2011) STING is a direct innate immune sensor of cyclic di-GMP. Nature 478:515–518
Sun L, Wu J, Du F, Chen X, Chen ZJ (2013) Cyclic GMP-AMP synthase is a cytosolic DNA sensor that activates the type I interferon pathway. Science 339:786–791
Wu J, Sun L, Chen X, Du F, Shi H, Chen C et al (2013) Cyclic GMP-AMP is an endogenous second messenger in innate immune signaling by cytosolic DNA. Science 339:826–830
Ablasser A, Goldeck M, Cavlar T, Deimling T, Witte G, Röhl I et al (2013) cGAS produces a 2′-5′-linked cyclic dinucleotide second messenger that activates STING. Nature 498(7454): 380–384
Sadler AJ, Williams BRG (2008) Interferon-inducible antiviral effectors. Nat Cell Biol 8:559–568
Lamkanfi M, Dixit VM (2012) Inflammasomes and their roles in health and disease. Annu Rev Cell Dev Biol 28:137–161
Hornung V, Ablasser A, Charrel-Dennis M, Bauernfeind F, Horvath G, Caffrey DR et al (2009) AIM2 recognizes cytosolic dsDNA and forms a caspase-1-activating inflammasome with ASC. Nature 458:514–518
Rathinam VAK, Vanaja SK, Fitzgerald KA (2012) Regulation of inflammasome signaling. Nat Immunol 13:333–342
Yoneyama M, Suhara W, Fukuhara Y, Sato M, Ozato K, Fujita T (1996) Autocrine amplification of type I interferon gene expression mediated by interferon stimulated gene factor 3 (ISGF3). J Biochem 120:160–169
Krieg AM (2002) CpG motifs in bacterial DNA and their immune effects. Annu Rev Immunol 20:709–760
Schmidt A, Schwerd T, Hamm W, Hellmuth JC, Cui S, Wenzel M et al (2009) 5′-triphosphate RNA requires base-paired structures to activate antiviral signaling via RIG-I. Proc Natl Acad Sci U S A 106:12067–12072
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
Kameyama, T., Takaoka, A. (2014). Characterization of Innate Immune Signalings Stimulated by Ligands for Pattern Recognition Receptors. In: Shiozawa, S. (eds) Arthritis Research. Methods in Molecular Biology, vol 1142. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0404-4_3
Download citation
DOI: https://doi.org/10.1007/978-1-4939-0404-4_3
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-0403-7
Online ISBN: 978-1-4939-0404-4
eBook Packages: Springer Protocols