Mechanisms of allergy and clinical immunologyProteome analysis of mast cell releasates reveals a role for chymase in the regulation of coagulation factor XIIIA levels via proteolytic degradation
Section snippets
Methods
For detailed methods, including the mice used, experimental protocols and procedures, and statistical analysis, please see the Methods section in this article's Online Repository at www.jacionline.org.
Proteome profiling of mast cell releasates
To define the BMCMC and PCMC releasates following IgE-mediated activation, we used a mass spectrometry shotgun proteomics approach. Specifically, IgE-2, 4-dinitrophenyl–sensitized BMCMCs and PCMCs were stimulated with 2, 4-dinitrophenyl-human serum albumin antigen for 6 hours. Then, the supernatants were collected, concentrated, and subjected to LC-MS/MS to identify the IgE-mediated differentially produced proteins from these mast cells (Fig 1, A). We identified 91 proteins that were unique to
Discussion
Accumulating in vitro and in vivo evidence indicates that mast cell functions may be much broader than their well-defined contribution to IgE-associated immune responses. In this study, we characterized mast cell releasates to further understand the contribution of mast cells to health and disease. For this purpose, we used proteomics as an unbiased approach to characterize mast cell secretomes. Previous studies used mass spectrometry analysis to dissect signaling pathways that are involved in
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2020, ImmunityCitation Excerpt :To aid future studies into mast cell functions, comprehensive proteome data are a valuable resource. Proteome analyses of mast cells have been previously reported for proteins released from mouse mast cells (Shubin et al., 2017), for human short-term cultured skin mast cells (Gschwandtner et al., 2017), for phosphoproteins (Cao et al., 2007), and for lipid rafts (Freitas Filho et al., 2019) in cultured rodent mast cells. Here, we isolated primary human connective tissue mast cells from skin and fat from healthy individuals and analyzed the mast cell proteome by quantitative mass spectrometry.
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2019, ImmunityCitation Excerpt :To investigate some of the identified candidate genes, we focused on FGL2. MCs release numerous soluble mediators, including proteases, growth factors, cytokines, and chemokines (Shubin et al., 2017). Our screen identified FGL2, which in its soluble form is mainly linked to Tregs and has suppressor activities against Th1 and Th17 cells, but not against Th2 cells (Joller et al., 2014).
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2019, BloodCitation Excerpt :Although we observe that serotonin is specifically important for DENV-induced platelet activation, MCs make other products that influence hemostasis. Chymase, a MC-specific protease, can prolong bleeding time by affecting the coagulation cascade,81 but is not required for DENV-induced thrombocytopenia, as antagonism of serotonin receptors or specific ablation of MC-serotonin synthase are sufficient to block thrombocytopenia. MC-stabilizing drugs, which can also restrict thrombocytopenia during DENV infection, are commercially available, are used in the treatment of asthma, and are currently being tested for treatment of dengue vascular leakage.
Research for this work was supported by grants for A.M.P. from the National Institutes of Health (NIH) (grant no. HL113351-01) and the American Heart Association (grant no. 12GRNT9680021), for R.G.J. from the NIH (grant no. 5R00HL103768-04), and for N.J.S. by a fellowship from the American Association of Immunologists (2015 AAI Careers in Immunology Fellowship). N.J.W. is supported, in part, by the National Center for Advancing Translational Sciences (grant no. KL2 TR000421), a component of the NIH. S.R.R. was supported by the Parker B. Francis Fellowship. M.A. was supported by the Swedish Research Council. This work is supported in part by the University of Washington's Proteomics Resource (grant no. UWPR95794).
Disclosure of potential conflict of interest: N. J. Shubin declares a grant from the American Academy of Immunologists. M. Abrink declares a grant from the Swedish Research Council. N. J. White declares grants/grants pending from the National Institutes of Health (NIH) and the US Department of Defense and is a board member, has provided consultancy to, and holds stock with Stasys Medical Corp. S. R. Reeves declares grants/grants pending from Parker B. Francis Foundation and the National Heart Lung and Blood Institute (NHLBI). R. G. James declares a grant from the NIH, NHLBI. T. Vaisar declares a grant from the NIH. A. M. Piliponsky declares grants from the NIH and the American Heart Association. The rest of the authors declare that they have no relevant conflicts of interest.