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TNF Production in Activated RBL-2H3 Cells Requires Munc13-4

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Abstract

Mast cell activation triggers intricate signaling pathways that promote the expression and/or release of a wide range of mediators including tumor necrosis factor (TNF; also known as TNFα). In this study, we investigated the connection between TNF secretion and TNF production, exploiting RBL-2H3 cells (a tumor analog of mucosal mast cells) that are depleted of Munc13-4, a crucial component of the mast cell exocytic machinery. We showed that antigen/IgE elicited robust TNF production in RBL-2H3 cells, but not in Munc13-4 knockout cells. The production defect was corrected when Munc13-4 was reintroduced into the knockout cell line, suggesting that the phenotype was not caused by any secondary effect derived from the knockout approach. Furthermore, pre-incubation of RBL-2H3 cells with R-7050, an antagonist of TNF receptor-dependent signaling, was shown to block TNF production without inhibiting TNF release. These observations provide fresh evidence for a robust feed-back loop to boost TNF production in activated mast cells.

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Funding

This work was financially supported by the National Institute of Allergy and Infectious Diseases Grant 1R15AI133430-01 to H.X. and S.S, and by the Mississippi INBRE, which was funded by an Institutional Development Award from the National Institute of General Medical Sciences under grant no. P20GM103476.

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Authors and Affiliations

  1. Department of Cell and Molecular Biology, School of Biological, Environmental, and Earth Sciences, University of Southern Mississippi, Hattiesburg, MS, 39406, USA

    Tolulope E. Ayo, Pratikshya Adhikari & Hao Xu

  2. Division of Fundamental Neurobiology, University Health Network, Toronto, ON, M5T 2S8, Canada

    Shuzo Sugita

  3. Department of Physiology, Faculty of Medicine, University of Toronto, Toronto, ON, M5S 1A8, Canada

    Shuzo Sugita

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  1. Tolulope E. Ayo
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Correspondence to Hao Xu.

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Ayo, T.E., Adhikari, P., Sugita, S. et al. TNF Production in Activated RBL-2H3 Cells Requires Munc13-4. Inflammation 43, 744–751 (2020). https://doi.org/10.1007/s10753-019-01161-4

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