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Distinctively Expressed Cytokines by Three Different Inflammation Cells and Their Interaction with Keratinocytes in Wound Healing

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Abstract

Inflammatory cells exert crucial influence on wound healing, while exploration is still desired to get further insight into the key factors that promote the process. In the present study, we performed comparative microarray data analysis of the three types of inflammatory cells isolated from skin wounds and focused on differentially expressed secreted factors. Gene Ontology enrichment and receptor analysis indicated that 11 genes of secreted factors in Ly6C+ inflammatory macrophages and 27 genes of secreted factors in neutrophils exhibited higher (≥ 5-fold) expression, and all these factors were considered as candidates with potentially important role in keratinocyte activation. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis revealed that TNF and IL17 signaling pathways were activated by Ly6C+ inflammatory macrophages secreted cytokines, while TGF-beta, HIF-1, NF-κB, and Rap 1 signaling pathways and pathways regulating pluripotency of stem cells were highly involved with neutrophils secreted cytokines. Taking TNF as an example, the source of the cytokine and its impact on keratinocytes were verified by immunofluorescence, gene knockout mice, and quantitative phosphoproteomics. Collectively, this study has indicated distinctively expressed cytokines in three inflammatory cells, which is helpful in identifying key factors in inducing keratinocyte signaling and in wound healing.

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Acknowledgements

We are grateful to Sobia Sadia for her assistance in language editing.

Funding

This work was supported by grants from Natural Science Foundation of China (No. 31371404, 31571429), Natural Science Foundation of Guangdong (2015A030311041), and Shenzhen Science and Technology Innovation Committee (GJHZ20150316160614842, JCY20160301150838144).

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Authors and Affiliations

  1. School of Life Sciences, Tsinghua University, Beijing, China

    Jingwen Wang & Xusheng Wang

  2. The Shenzhen Key Laboratory of Health Sciences and Technology, Graduate School at Shenzhen, Tsinghua University, Shenzhen, China

    Jingwen Wang, Xusheng Wang & Yaojiong Wu

  3. Tsinghua-Berkeley Shenzhen Institute (TBSI), Tsinghua University, Shenzhen, China

    Haiyan Chen & Yaojiong Wu

  4. Medical Key Laboratory of Health Toxicology of Shenzhen, Shenzhen Center for Disease Control and Prevention, Shenzhen, China

    Jianjun Liu

  5. Wound Healing Research Group, Department of Surgery, University of Alberta, Edmonton, AB, Canada

    Edward E. Tredget

  6. L406A, Tsinghua Campus, The University Town, Shenzhen, China

    Yaojiong Wu

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  1. Jingwen Wang
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  2. Xusheng Wang
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  4. Jianjun Liu
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Correspondence to Yaojiong Wu.

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All procedures were performed with the approval of the Animal Ethics Committee of Tsinghua University.

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Supplementary Figure 1

Gating and sorting of three types of inflammatory cells. Single cell suspensions derived from skin wound tissues were subjected to flow cytometry analysis. Plot was gated largely to avoid cell debrils (a). Residential macrophages were defined as F4/80+/CX3CR1+ cells, inflammatory macrophages were gated as F4/80+/Ly6C+ cells, and neutrophils were sorted for the F4/80-/Gr-1+ subset. Cells stained with corresponding isotype IgGs were used as a negative controls (b). (JPEG 10 kb)

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Wang, J., Wang, X., Chen, H. et al. Distinctively Expressed Cytokines by Three Different Inflammation Cells and Their Interaction with Keratinocytes in Wound Healing. Inflammation 40, 2151–2162 (2017). https://doi.org/10.1007/s10753-017-0655-9

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  • DOI: https://doi.org/10.1007/s10753-017-0655-9

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