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Mannan-binding lectin directly interacts with Toll-like receptor 4 and suppresses lipopolysaccharide-induced inflammatory cytokine secretion from THP-1 cells

Cellular & Molecular Immunology volume 8, pages 265–275 (2011)Cite this article

Abstract

Mannan-binding lectin (MBL) plays a key role in the lectin pathway of complement activation and can influence cytokine expression. Toll-like receptor 4 (TLR4) is expressed extensively and has been demonstrated to be involved in lipopolysaccharide (LPS)-induced signaling. We first sought to determine whether MBL exposure could modulate LPS-induced inflammatory cytokine secretion and nuclear factor-κB (NF-κB) activity by using the monocytoid cell line THP-1. We then investigated the possible mechanisms underlying any observed regulatory effect. Using ELISA and reverse transcriptase polymerase chain reaction (RT-PCR) analysis, we found that at both the protein and mRNA levels, treatment with MBL suppresses LPS-induced tumor-necrosis factor (TNF)-α and IL-12 production in THP-1 cells. An electrophoretic mobility shift assay and western blot analysis revealed that MBL treatment can inhibit LPS-induced NF-κB DNA binding and translocation in THP-1 cells. While the binding of MBL to THP-1 cells was evident at physiological calcium concentrations, this binding occurred optimally in response to supraphysiological calcium concentrations. This binding can be partly inhibited by treatment with either a soluble form of recombinant TLR4 extracellular domain or anti-TLR4 monoclonal antibody (HTA125). Activation of THP-1 cells by LPS treatment resulted in increased MBL binding. We also observed that MBL could directly bind to the extracellular domain of TLR4 in a dose-dependent manner, and this interaction could attenuate the binding of LPS to cell surfaces. Taken together, these data suggest that MBL may affect cytokine expression through modulation of LPS-/TLR-signaling pathways. These findings suggest that MBL may play an important role in both immune regulation and the signaling pathways involved in cytokine networks.

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Acknowledgements

This work was supported by the Natural Science Foundation of China (30972679). Special thanks are given to Professors Tianyun Wang, Wenming Yong and Weiren Dong for their dedicated revision of the paper.

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  1. Dr ZL Chen, Department of Immunology, Southern Medical University, Guangzhou 510515, China. E-mail: zhlchen@fimmu.com

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  1. Department of Immunology, Southern Medical University, Guangzhou, 510515, China

    Mingyong Wang, Yue Chen, Yani Zhang, Liyun Zhang, Xiao Lu & Zhengliang Chen

  2. Department of Medical Tests, Xinxiang Medical University, Xinxiang, 453003, China

    Mingyong Wang

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Wang, M., Chen, Y., Zhang, Y. et al. Mannan-binding lectin directly interacts with Toll-like receptor 4 and suppresses lipopolysaccharide-induced inflammatory cytokine secretion from THP-1 cells. Cell Mol Immunol 8, 265–275 (2011). https://doi.org/10.1038/cmi.2011.1

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