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Impairment of Immune Response against Dematiaceous Fungi in Card9 Knockout Mice

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Abstract

Dematiaceous fungi are a large group of pathogens that can cause a wide range of diseases in both immunocompetent and immunocompromised hosts. Based on our previous finding of caspase recruitment domain-containing protein 9 (CARD9) mutations in patients with subcutaneous phaeohyphomycosis caused by Phialophora verrucosa (P. verrucosa), we further investigated the exact role of CARD9 in the pathogenesis of phaeohyphomycosis using Card9 knockout (Card9 KO) mice. We showed that Card9 KO mice are profoundly susceptible to P. verrucosa infection compared with wild-type mice, reflected by significantly more severe footpad swelling, higher fungal burden, lower survival, and systemic dissemination. The inability of Card9 KO mice to control P. verrucosa infection was associated with lack of Th17 differentiation and reduction of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and IL-17A levels in footpad homogenates. In vitro experiments showed a defect of fungal conidia killing and pro-inflammatory cytokine production in Card9 KO bone marrow-derived macrophages and dendritic cells. Furthermore, ex vivo coculture and in vitro T cell differentiation assay demonstrated that Card9 signaling pathway acts indispensably on differentiation of Th17 cells. In conclusion, our findings suggest that CARD9 mediate the innate immune and Th17-mediated adaptive immune responses against dematiaceous fungal infections at the early stage of infection.

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Acknowledgments

We thank Dr. Xin Lin for kindly providing the Card9 knockout mice; we thank Zhe Wan at the Department of Dermatology, Peking University First Hospital, for isolation and identification of the pathogen; Hao Zhang at the Department of Dermatology, Peking University First Hospital, for assistance with image processing; and Huihui Liu, Department of Hematology, Peking University First Hospital, for technical advice on the in vitro functional analyses of BMDCs. The authors are grateful for the financial support of National Natural Science Foundation of China (Grant 81472890) and Peking University (PKU) 985 Special Funding for Collaborative Research with PKU Hospitals (Grant 2014-1-3).

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    Authors and Affiliations

    1. Department of Dermatology, Peking University First Hospital, Beijing, China

      Weiwei Wu, Ruijun Zhang, Xiaowen Wang, Yinggai Song & Ruoyu Li

    2. Research Center for Medical Mycology, Peking University, Beijing, China

      Weiwei Wu, Ruijun Zhang, Xiaowen Wang, Yinggai Song & Ruoyu Li

    3. Beijing Key Laboratory of Molecular Diagnosis on Dermatoses, Beijing, China

      Weiwei Wu, Ruijun Zhang, Xiaowen Wang, Yinggai Song & Ruoyu Li

    4. Department of Immunology, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, China

      Zhengyang Liu & Wenling Han

    5. Peking University Center for Human Disease Genomics, Beijing, China

      Zhengyang Liu & Wenling Han

    6. Key Laboratory of Medical Immunology, Ministry of Health, Beijing, China

      Zhengyang Liu & Wenling Han

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    1. Weiwei Wu
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    Correspondence to Ruoyu Li.

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    All procedures performed in studies involving animals were in accordance with institutional guidelines and were approved by the Institutional Ethics Committee of Peking University First Hospital in accordance with the National Institutes of Health Animal Care Guidelines.

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    Weiwei Wu and Ruijun Zhang have contributed equally to this work.

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    Wu, W., Zhang, R., Wang, X. et al. Impairment of Immune Response against Dematiaceous Fungi in Card9 Knockout Mice. Mycopathologia 181, 631–642 (2016). https://doi.org/10.1007/s11046-016-0029-0

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