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The adaptor protein CARD9 is required for innate immune responses to intracellular pathogens

Nature Immunology volume 8, pages 198–205 (2007)Cite this article

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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Figure 1: Characterization of the mouse Card9 expression profile.
Figure 2: CARD9 is required for the production of proinflammatory cytokines after microbial mimetic stimulation.
Figure 3: Signaling defects of CARD9-deficient macrophages.
Figure 4: Induction of proinflammatory cytokines and activation of p38 and Jnk require CARD9 during intracellular bacterial and viral infection.
Figure 5: CARD9 acts in synergy with Nod2 in the activation of p38 but not of NF-κB.
Figure 6: CARD9 associates with Nod2 in innate immune responses.
Figure 7: CARD9 is required for the early production of proinflammatory cytokines in response to intracellular bacteria infection.
Figure 8: Card9−/− mice are impaired in the clearance of intracellular bacteria.

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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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Authors and Affiliations

  1. Department of Molecular and Cellular Oncology, University of Texas, M.D. Anderson Cancer Center, Houston, 77030, Texas, USA

    Yen-Michael S Hsu, Yun You, Hongxiu Li & Xin Lin

  2. Department of Immunology, University of Texas, M.D. Anderson Cancer Center, Houston, 77030, Texas, USA

    Yongliang Zhang, Omar Duramad, Xiao-Feng Qin & Chen Dong

  3. The CBR Institute for Biomedical Research, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Donghai Wang

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  1. Yen-Michael S Hsu
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Contributions

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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Correspondence to Xin Lin.

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The authors declare no competing financial interests.

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)

Supplementary Methods (PDF 69 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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