Immunomodulatory Functions of Glycolipids from Pathogens

Carla Guenther, Miyuki Watanabe, Sho Yamasaki

    Research output: Chapter in Book/Report/Conference proceedingChapterScientificpeer-review

    Abstract

    The cell envelopes of pathogens comprise a wealth of unique glycolipids, which are important modulators of the host immune responses during infection and in some cases have been used as adjuvants. Despite this abundant basic knowledge, the identities of the host immune receptors for mycobacterial lipids have long been elusive (Ishikawa et al., Trends Immunol 38:66–76, 2017). We describe the method of how to isolate glycolipids from microorganisms and how to analyze the glycolipids’ potential to activate reporter cells and bone marrow-derived dendritic cells (BMDCs), such as surface marker expression and reactive oxygen species (ROS) production. Additionally, we outline an in vitro BMDC/T cell coculture model to investigate functional consequences of leukocyte activation, such as cytokine production. In this chapter, we provide a guide for extracting glycolipids from microorganisms and how to use them to activate leukocytes. We also present methods on how to generate and activate reporter cells, as well as BMDCs and how to set up BMDC/T cell cocultures. We further outline how to generate samples and how to analyze the immunomodulatory effect glycolipid exposure has on these cells, via flow cytometry, ROS production assays and ELISA.

    Original languageEnglish
    Title of host publicationGlycolipids
    Subtitle of host publicationMethods and Protocols
    EditorsKazuya Kabayama, Jin-ichi Inokuchi
    PublisherHumana Press
    Pages23-31
    Number of pages9
    ISBN (Electronic)978-1-0716-2910-9
    ISBN (Print)978-1-0716-2909-3
    DOIs
    Publication statusPublished - 2023
    Publication typeA3 Book chapter

    Publication series

    NameMethods in Molecular Biology
    Volume2613
    ISSN (Print)1064-3745
    ISSN (Electronic)1940-6029

    Keywords

    • BMDC generation
    • BMDC/T cell co-culture
    • Glycolipid extraction
    • Myeloid cell activation assay
    • Reporter cell assay
    • ROS production assay

    Publication forum classification

    • Publication forum level 1

    ASJC Scopus subject areas

    • Molecular Biology
    • Genetics

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