@inbook{41916c8fd5c741b38c8ab7e2a1179ecf,
title = "Immunomodulatory Functions of Glycolipids from Pathogens",
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{\textquoteright} 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.",
keywords = "BMDC generation, BMDC/T cell co-culture, Glycolipid extraction, Myeloid cell activation assay, Reporter cell assay, ROS production assay",
author = "Carla Guenther and Miyuki Watanabe and Sho Yamasaki",
note = "Publisher Copyright: {\textcopyright} 2023, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.",
year = "2023",
doi = "10.1007/978-1-0716-2910-9_3",
language = "English",
isbn = "978-1-0716-2909-3",
series = "Methods in Molecular Biology",
publisher = "Humana Press",
pages = "23--31",
editor = "Kabayama, {Kazuya } and Jin-ichi Inokuchi",
booktitle = "Glycolipids",
address = "United States",
}