Investigating IL-1β Secretion Using Real-Time Single-Cell Imaging.

Catherine Diamond, James Bagnall, David Spiller, Michael White, Alessandra Mortellaro, Pawel Paszek, David Brough

    Research output: Chapter in Book/Conference proceedingChapterpeer-review

    Abstract

    The pro-inflammatory cytokine interleukin (IL)-1β is an important mediator of the inflammatory response. In order to perform its role in the inflammatory cascade, IL-1β must be secreted from the cell, yet it lacks a signal peptide that is required for conventional secretion, and the exact mechanism of release remains undefined. Conventional biochemical methods have limited the investigation into the processes involved in IL-1β secretion to population dynamics, yet heterogeneity between cells has been observed at a single-cell level. Here, greater sensitivity is achieved with the use of a newly developed vector that codes for a fluorescently labelled version of IL-1β. Combining this with real-time single-cell confocal microscopy using the methods described here, we have developed an effective protocol for investigating the mechanisms of IL-1β secretion and the testing of the hypothesis that IL-1β secretion requires membrane permeabilisation.
    Original languageEnglish
    Title of host publicationNLR proteins
    Subtitle of host publicationMethods and protocols
    EditorsFrancesco Di Virgilio, Pelegrín Pablo
    PublisherSpringer Nature
    Pages75-88
    Number of pages13
    ISBN (Electronic)978-1-4939-3566-6
    ISBN (Print)978-1-4939-3564-2
    DOIs
    Publication statusPublished - 2016

    Publication series

    NameMethods in Molecular Biology
    PublisherSpringer New York
    Volume1417
    ISSN (Print)1064-3745

    Research Beacons, Institutes and Platforms

    • Dementia@Manchester

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