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Extracellular calcium elicits feedforward regulation of the Toll-like receptor-triggered innate immune response

Cellular & Molecular Immunology volume 14, pages 180–191 (2017)Cite this article

Abstract

Despite the expanding knowledge on feedback regulation of Toll-like receptor (TLR) signaling, the feedforward regulation of TLR signaling for the proper innate response to invading microbes is not fully understood. Here, we report that extracellular calcium can coordinate the activation of the small GTPases Ras and Ras-proximate-1 (Rap1) upon TLR stimulation which favors activation of macrophages through a feedforward mechanism. We show that different doses of TLR agonists can trigger different levels of cytokine production, which can be potentiated by extracellular calcium but are impaired by the chelating reagent ethylene glycol tetraacetic acid (EGTA) or by knockdown of stromal interaction molecule 1 (STIM1). Upon TLR engagement, GTP-bound Ras levels are increased and GTP-bound Rap1 is decreased, which can be reversed by EGTA-mediated removal of extracellular calcium. Furthermore, we demonstrate that Rap1 knockdown rescues the inhibitory effects of EGTA on the TLR-triggered innate response. Examination of the TLR signaling pathway reveals that extracellular calcium may regulate the TLR response via feedforward activation of the extracellular signal-regulated kinase signaling pathway. Our data suggest that an influx of extracellular calcium, mediated by STIM1-operated calcium channels, may transmit the information about the intensity of extracellular TLR stimuli to initiate innate responses at an appropriate level. Our study may provide mechanistic insight into the feedforward regulation of the TLR-triggered innate immune response.

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Acknowledgements

This work was supported by grants from the National Key Basic Research Program of China (2010CB911903 and 2013CB530502), the National Natural Science Foundation of China (81172851, 81222039, 31270944, and 31370902), and the National High Technology Research and Development Program (2012AA020900). We thank Dr. Xingguang Liu for helpful discussion and assistance with manuscript writing, and Ms. Mei Jin and Ms. Hao Shen for their excellent technical assistance.

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Author notes

  1. Songqing Tang and Taoyong Chen: Songqing Tang and Taoyong Chen contributed equally to this work.

  2. Dr. J Wang, Institute of Immunology, Zhejiang University School of Medicine, 866 Yuhangtang Road, Hangzhou 310058, China. E-mail: jlwang@zju.edu.cn

Authors and Affiliations

  1. Institute of Immunology, Zhejiang University School of Medicine, Hangzhou, 310058, China

    Songqing Tang, Zhou Yu, Bin Xie, Xuetao Cao & Jianli Wang

  2. National Key Laboratory of Medical Immunology & Institute of Immunology, Second Military Medical University, Shanghai, 200433, China

    Songqing Tang, Taoyong Chen, Mingjin Yang, Bin Xie, Cheng Qian, Sheng Xu, Nan Li & Xuetao Cao

  3. National Key Laboratory of Medical Molecular Biology & Department of Immunology, Chinese Academy of Medical Sciences, Beijing, 100005, China

    Mingjin Yang, Lei Wang & Xuetao Cao

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Tang, S., Chen, T., Yang, M. et al. Extracellular calcium elicits feedforward regulation of the Toll-like receptor-triggered innate immune response. Cell Mol Immunol 14, 180–191 (2017). https://doi.org/10.1038/cmi.2015.59

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