NOD2 is a negative regulator of Toll-like receptor 2–mediated T helper type 1 responses

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Abstract

The mechanism by which mutations in CARD15, which encodes nucleotide-binding oligomerization domain 2 (NOD2), cause Crohn disease is poorly understood. Because signaling via mutated NOD2 proteins leads to defective activation of the transcription factor NF-κB, one proposal is that mutations cause deficient NF-κB-dependent T helper type 1 (TH1) responses and increased susceptibility to infection. However, this idea is inconsistent with the increased TH1 responses characteristic of Crohn disease. Here we used Card15−/− mice to show that intact NOD2 signaling inhibited Toll-like receptor 2–driven activation of NF-κB, particularly of the NF-κB subunit c-Rel. Moreover, NOD2 deficiency or the presence of a Crohn disease–like Card15 mutation increased Toll-like receptor 2–mediated activation of NF-κB–c-Rel, and TH1 responses were enhanced. Thus, CARD15 mutations may lead to disease by causing excessive TH1 responses.

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Figure 1: Splenocytes from Card15−/− mice show enhanced IL-12 and IFN-γ production after stimulation with PGN.
Figure 2: Splenocytes from wild-type mice but not Card15−/− mice show decreased TH1 cytokine production after costimulation with TLR2 and NOD2 agonists.
Figure 3: TH1 cytokine production and TLR2 and IL-12R expression by purified spleen subpopulations from wild-type and Card15−/− mice.
Figure 4: Wild-type spleen macrophages show dose-dependent suppression of TLR2-induced IL-12 production mediated by MDP costimulation and a lack of effect of costimulation on IL-12 production induced by stimulants of other TLRs.
Figure 5: Card15−/− mice show enhanced IL-12 production after systemic administration of PGN.
Figure 6: Stimulation of splenic adherent cells with PGN enhances translocation of c-Rel into the nucleus.
Figure 7: Neutralization of c-Rel by siRNA inhibits TLR2-mediated IL-12 production by splenic CD11b+ cells from Card15−/− mice.
Figure 8: CD11b+ cells from Card15−/− mice expressing transfected Card15 with a frameshift mutation show enhanced PGN-induced IL-12 production.

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Acknowledgements

We thank J.T. Rosenbaum and M.P. Davey (Casey Eye Institute, Oregon Health and Science University) for providing us with the plasmid expressing mouse Card15; and C. Ma and S. Fichtner (Laboratory of Host Defenses, National Institute of Allergy and Infectious Diseases) for comments and technical assistance. Supported in part by the Cancer Center CORE (P30 CA21765) and the American Lebanese Associated Charities.

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Correspondence to Warren Strober.

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Watanabe, T., Kitani, A., Murray, P. et al. NOD2 is a negative regulator of Toll-like receptor 2–mediated T helper type 1 responses. Nat Immunol 5, 800–808 (2004) doi:10.1038/ni1092

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