Abstract
Nod-like receptors (NLRs) are intracellular sensors that respond to a variety of pathogen and intracellular danger signals to induce innate immune responses. NLRC5 has recently been identified to be an important regulator of NF-κB, type I interferon (IFN) and inflammasome signaling pathways, but the in vivo function and mechanisms of NLRC5 remain to be defined. Here, we describe the generation and characterization of NLRC5 knockout mice. We show that induction of NLRC5 expression by Toll-like receptor (TLR) ligand or cytokine stimulation requires the signal transducers and activators of transcription (Stat)1-mediated signaling pathway. NLRC5 ablation reduces MHC class I expression, and enhances IKK and IRF3 phosphorylation in response to TLR stimulation or viral infection. Consistent with these observations, we found that NLRC5 deficiency enhanced IL-6 and IFN-β production in mouse embryonic fibroblasts (MEFs), peritoneal macrophages and bone marrow-derived macrophages (BMMs), but not bone marrow-derived dendritic cells (BMDCs) after LPS stimulation or vesicular stomatitis virus (VSV) infection. Furthermore, we found that NLRC5-deficient mice produced higher amounts of IL-6 and IFN-β in the sera when they were challenged with LPS or infected with VSV. Taken together, these results provide in vivo evidence that NLRC5 plays critical roles in MHC class I expression, innate immune signaling and antiviral innate immune responses, thus serving as an important target for modulating innate immune signaling and regulation.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Takeuchi O, Akira S . Pattern recognition receptors and inflammation. Cell 2010; 140:805–820.
Schroder K, Tschopp J . The inflammasomes. Cell 2010; 140:821–832.
Kawai T, Akira S . The role of pattern-recognition receptors in innate immunity: update on Toll-like receptors. Nat Immunol 2010; 11:373–384.
Wilkins C, Gale M Jr . Recognition of viruses by cytoplasmic sensors. Curr Opin Immunol 2010; 22:41–47.
Chiu YH, Macmillan JB, Chen ZJ . RNA polymerase III detects cytosolic DNA and induces type I interferons through the RIG-I pathway. Cell 2009; 138:576–591.
Ablasser A, Bauernfeind F, Hartmann G, Latz E, Fitzgerald KA, Hornung V . RIG-I-dependent sensing of poly(dA:dT) through the induction of an RNA polymerase III-transcribed RNA intermediate. Nat Immunol 2009; 10:1065–1072.
Zhang Z, Yuan B, Bao M, Lu N, Kim T, Liu YJ . The helicase DDX41 senses intracellular DNA mediated by the adaptor STING in dendritic cells. Nat Immunol 2011; 12:959–965.
Unterholzner L, Keating SE, Baran M, et al. IFI16 is an innate immune sensor for intracellular DNA. Nat Immunol 2010; 11:997–1004.
Inohara N, Nunez G . NODs: intracellular proteins involved in inflammation and apoptosis. Nat Rev Immunol 2003; 3:371–382.
Kufer TA, Sansonetti PJ . NLR functions beyond pathogen recognition. Nat Immunol 2011; 12:121–128.
Moore CB, Bergstralh DT, Duncan JA, et al. NLRX1 is a regulator of mitochondrial antiviral immunity. Nature 2008; 451:573–577.
Xia X, Cui J, Wang HY, et al. NLRX1 negatively regulates TLR-induced NF-kappaB signaling by targeting TRAF6 and IKK. Immunity 2011; 34:843–853.
Allen IC, Moore CB, Schneider M, et al. NLRX1 protein attenuates inflammatory responses to infection by interfering with the RIG-I-MAVS and TRAF6-NF-kappa B signaling pathways. Immunity 2011; 34:854–865.
Tattoli I, Carneiro LA, Jehanno M, et al. NLRX1 is a mitochondrial NOD-like receptor that amplifies NF-kappa B and JNK pathways by inducing reactive oxygen species production. EMBO Rep 2008; 9:293–300.
Jounai N, Kobiyama K, Shiina M, Ogata K, Ishii KJ, Takeshita F . NLRP4 negatively regulates autophagic processes through an association with Beclin1. J Immunol 2011; 186:1646–1655.
Fiorentino L, Stehlik C, Oliveira V, Ariza ME, Godzik A, Reed JC . A novel PAAD-containing protein that modulates NF-kappa B induction by cytokines tumor necrosis factor-alpha and interleukin-1 beta. J Biol Chem 2002; 277:35333–35340.
Cui J, Zhu L, Xia XJ, et al. NLRC5 negatively regulates the NF-kappa B and type I interferon signaling pathways. Cell 2010; 141:483–496.
Benko S, Magalhaes JG, Philpott DJ, Girardin SE . NLRC5 limits the activation of inflammatory pathways. J Immunol 2010; 185:1681–1691.
Kuenzel S, Till A, Winkler M, et al. The nucleotide-binding oligomerization domain-like receptor NLRC5 is involved in IFN-dependent antiviral immune responses. J Immunol 2010; 184:1990–2000.
Neerincx A, Lautz K, Menning M, et al. A role for the human nucleotide-binding domain, leucine-rich repeat-containing family member NLRC5 in antiviral responses. J Biol Chem 2010; 285:26223–26232.
Davis BK, Roberts RA, Huang MT, et al. Cutting edge: NLRC5-dependent activation of the inflammasome. J Immunol 2011; 186:1333–1337.
Meissner TB, Li A, Biswas A, et al. NLR family member NLRC5 is a transcriptional regulator of MHC class I genes. Proc Natl Acad Sci USA 2010; 107:13794–13799.
Kumar H, Pandey S, Zou J, et al. NLRC5 deficiency does not influence cytokine induction by virus and bacteria infections. J Immunol 2011; 186:994–1000.
O'Shea JJ, Gadina M, Schreiber RD . Cytokine signaling in 2002: new surprises in the Jak/Stat pathway. Cell 2002; 109 Suppl:S121–S131.
Shuai K, Liu B . Regulation of JAK-STAT signalling in the immune system. Nat Rev Immunol 2003; 3:900–911.
LeibundGut-Landmann S, Waldburger JM, Krawczyk M, et al. Mini-review: specificity and expression of CIITA, the master regulator of MHC class II genes. Eur J Immunol 2004; 34:1513–1525.
Wright KL, Ting JP . Epigenetic regulation of MHC-II and CIITA genes. Trends Immunol 2006; 27:405–412.
Honda K, Taniguchi T . IRFs: master regulators of signalling by Toll-like receptors and cytosolic pattern-recognition receptors. Nat Rev Immunol 2006; 6:644–658.
Ajibade AA, Wang Q, Cui J, et al. TAK1 negatively regulates NF-kappaB and p38 MAP kinase activation in Gr-1+CD11b+ neutrophils. Immunity 2012; 36:43–54.
Kawagoe T, Takeuchi O, Takabatake Y, et al. TANK is a negative regulator of Toll-like receptor signaling and is critical for the prevention of autoimmune nephritis. Nat Immunol 2009; 10:965–972.
Han J, Ulevitch RJ . Limiting inflammatory responses during activation of innate immunity. Nat Immunol 2005; 6:1198–1205.
Kanzler H, Barrat FJ, Hessel EM, Coffman RL . Therapeutic targeting of innate immunity with Toll-like receptor agonists and antagonists. Nat Med 2007; 13:552–559.
Li HY, Liu H, Wang CH, et al. Deactivation of the kinase IKK by CUEDC2 through recruitment of the phosphatase PP1. Nat Immunol 2008; 9:533–541.
Rebsamen M, Vazquez J, Tardivel A, Guarda G, Curran J, Tschopp J . NLRX1/NOD5 deficiency does not affect MAVS signalling. Cell Death Differ 2011; 18:1387.
Acknowledgements
We would like to thank Dr Adebusola A Ajibade (The Methodist Hospital Research Institute) for critical reading of this manuscript, and Drs Margaret A Goodell and Katherine Yudeh King (Baylor College of Medicine) for providing us with Stat1−/− mice. This work was in part supported by Grants from National Institutes of Health (NIH) (CA090327, CA101795, CA121191, CA116408, CA094327 and DA030338), Cancer Research Institute, and The Methodist Hospital Research Institute. Yanzheng Tong was a recipient of The China Scholarship Council (CSC).
Author information
Authors and Affiliations
Corresponding author
Additional information
( Supplementary information is linked to the online version of the paper on the Cell Research website.)
Supplementary information
Supplementary information, Figure S1 (download PDF )
Induction of NLRC5 expression by the cytokines in the cell culture supernatant after LPS stimulation (PDF 633 kb)
Supplementary information, Figure S2 (download PDF )
Southern blotting analysis of WT and NLRC5+/− embryonic stem cell clones by (A) 5′ probe 1 and (B) 3′ probe 2 shown in Figure 2A. (PDF 41 kb)
Supplementary information, Figure S3 (download PDF )
Enhanced TNF-α production in NLRC5-deficient MEF cells (PDF 264 kb)
Supplementary information, Figure S4 (download PDF )
NLRC5 deletion has little or no effect on cytokine production in BMDCs. (PDF 110 kb)
Supplementary information, Figure S5 (download PDF )
Loss of NLRC5 has little or no effect on TNF-α and IL-1β production in peritoneal macrophages and BMMs. (PDF 273 kb)
Supplementary information, Figure S6 (download PDF )
Much more differences in IL-6 and TNF-α release between NLRC5−/− and WT peritoneal macrophages cells were observed only when low, but not high, doses of LPS were used for treatment. (PDF 116 kb)
Supplementary information, Figure S7 (download PDF )
Loss of NLRC5 has little or no effect on mice survival or virus titer of plasma after VSV-eGFP infection in vivo. (PDF 46 kb)
Rights and permissions
About this article
Cite this article
Tong, Y., Cui, J., Li, Q. et al. Enhanced TLR-induced NF-κB signaling and type I interferon responses in NLRC5 deficient mice. Cell Res 22, 822–835 (2012). https://doi.org/10.1038/cr.2012.53
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/cr.2012.53
Keywords
This article is cited by
-
NLRC5/CITA expression correlates with efficient response to checkpoint blockade immunotherapy
Scientific Reports (2021)
-
Classical MHC expression by DP thymocytes impairs the selection of non-classical MHC restricted innate-like T cells
Nature Communications (2021)
-
NLRC5 regulates expression of MHC-I and provides a target for anti-tumor immunity in transmissible cancers
Journal of Cancer Research and Clinical Oncology (2021)
-
A new molecular classification to drive precision treatment strategies in primary Sjögren’s syndrome
Nature Communications (2021)
-
NLRC5: new cancer buster?
Molecular Biology Reports (2020)
