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Protein Profiling of the Secretome of FcεRI Activated RBL-2H3.1 Cells

Author(s): Esmaeil Sadroddiny | Jafar Ai | Kathleen Carroll | Trong Khoa Pham | Phillip Wright | Ashutosh Pathak | Birgit Helm

Journal: Iranian Journal of Immunology
ISSN 1735-1383

Volume: 9;
Issue: 1;
Start page: 1;
Date: 2012;
Original page

Keywords: Basophil | FcεRI | IgE | Mast Cell | RBL-2H3.1 | Secretome

Background: Secretory proteins of IgE receptor activated mast cells and basophils playa pivotal role in the generation of immediate and long term immune responses in allergyand type I hypersensitivity. Objective: The present study aims to generate a 2-D mapand profile of proteins secreted from a high secretory variant of the rat basophilic leukemiacell line, RBL-2H3.1, which in view of the difficulty associated with gainingadequate numbers of pure primary mast cell and basophiles, represents an acceptedmodel system for the study and standardization of the methodology to characterize thesecretome of these cell types. Methods: A 2-D map of secretory proteins was generatedby 2-D PAGE and a shotgun mass spectrometric approach carried out for protein identification. Results: Study resulted into identification of 299 proteins released from restingand IgE receptor activated RBL-2H3.1 cells after 90 s, 30 min and 3 h antigen challenge.Further sequence analysis identified ~53% of total proteins as secretory proteinswhich could be attributed to classical and non-classical secretory pathways. Additionally,functional classification of classic secretory proteins verified the presence of proteinsbelonged to cytokines, receptors, membrane proteins, lysosomal proteins and proteinsassociated with specific sub-cellular localizations such as endoplasmic reticulum,mitochondria, nucleus, cytoplasm and ribosome. According to this data the presence ofsome secretory proteins such as cytokines (e.g. MCP-2, PF-4, CSF-1 and TGF-β1) areall subject to Ag challenge which may point to their importance toward pathogenesis inallergic diseases. Conclusion: In view of both a beneficial and adverse role of mast cellmediators in health and disease, an identification of temporal changes in the secretorypattern may form the basis for future tailor made intervention strategies that may enableus to harvest the therapeutic potential inherent in mast cell exocytosis while inhibiting/attenuating negative outcomes.

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