Abstract
The caspase-recruitment domain–containing adaptor protein CARD9 regulates the innate signaling responses to fungal infection. Here we show that CARD9 is required for innate immune responses against intracellular pathogens. We generated Card9−/− mice and found that CARD9-deficient macrophages had defects in activation of the kinases p38 and Jnk but not of transcription factor NF-κB after bacterial and viral infection. CARD9-deficient mice failed to clear infection and showed altered cytokine production after challenge with Listeria monocytogenes. In wild-type cells, we found CARD9 inducibly associated with both the intracellular 'biosensor' Nod2 and the serine-threonine kinase RICK. Our data demonstrate that CARD9 has a critical function in Nod2-mediated activation of p38 and Jnk in innate immune responses to intracellular pathogens.
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Acknowledgements
We thank K. Schluns (M.D. Anderson Cancer Center) for reagents, and G. Nunez (University of Michigan) and G. Cheng (University of California at Los Angeles) for hemagglutinin-tagged Nod2, Myc-tagged RICK and Flag-tagged RICK. Supported by the National Institutes of Health, the Leukemia and Lymphoma Society (X.L.), the Cancer Research Institute (X.L.) and the MD Anderson Cancer Center (Y.L.).
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Y.-M.S.H., Y.Z., Y.Y., D.W., H.L. and O.D. performed the experiments. X.-F.Q., C.D. and X.L. analyzed the data.
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Supplementary information
Supplementary Fig. 1
Generation of Card9−/− mice. (PDF 224 kb)
Supplementary Fig. 2
CARD9 expression in lung and specific lymphoid organs. (PDF 154 kb)
Supplementary Fig. 3
CARD9 is required for the production of TNF upon selective microbial mimetic stimulation. (PDF 159 kb)
Supplementary Fig. 4
Microbial mimetic–stimulated Card9 deficient macrophages exhibit a dosage-dependent reduction in IL-6 production. (PDF 169 kb)
Supplementary Fig. 5
LPS and FSL-1-induced cytokine production is normal in CARD9-deficient macrophages. (PDF 188 kb)
Supplementary Fig. 6
MAPK activation and IκBα degradation are not altered in CARD9-deficient macrophages upon LPS or pure-LPS stimulation. (PDF 225 kb)
Supplementary Fig. 7
Virus-induced IL-6 and TNF transcription is defective in CARD9-deficient macrophages. (PDF 171 kb)
Supplementary Fig. 8
CARD9 overexpression leads to enhanced p38 and Jnk phosphorylation but not NF-κB activation. (PDF 225 kb)
Supplementary Fig. 9
Working model showing that CARD9 mediates Nod2-induced innate immune response through p38 and JNK activation. (PDF 91 kb)
Supplementary Fig. 10
Card9 deficiency leads to sustained spleen enlargement and inflammation upon listeria infection. (PDF 197 kb)
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Hsu, YM., Zhang, Y., You, Y. et al. The adaptor protein CARD9 is required for innate immune responses to intracellular pathogens. Nat Immunol 8, 198–205 (2007). https://doi.org/10.1038/ni1426
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DOI: https://doi.org/10.1038/ni1426
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