Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023:2613:23-31.
doi: 10.1007/978-1-0716-2910-9_3.

Immunomodulatory Functions of Glycolipids from Pathogens

Carla Guenther  1   2   3 , Miyuki Watanabe  2   3 , Sho Yamasaki  4   5   6   7
Affiliations

Immunomodulatory Functions of Glycolipids from Pathogens

Carla Guenther et al. Methods Mol Biol. 2023.

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.

Keywords: BMDC generation; BMDC/T cell co-culture; Glycolipid extraction; Myeloid cell activation assay; ROS production assay; Reporter cell assay.

PubMed Disclaimer

References

    1. Ishikawa E, Mori D, Yamasaki S (2017) Recognition of mycobacterial lipids by immune receptors. Trends Immunol 38:66–76 - DOI
    1. Chiffoleau E (2018) C-type lectin-like receptors as emerging orchestrators of sterile inflammation represent potential therapeutic targets. Front Immunol 9:227 - DOI
    1. Yamasaki S, Ishikawa E, Sakuma M et al (2008) Mincle is an ITAM-coupled activating receptor that senses damaged cells. Nat Immunol 9:1179–1188 - DOI
    1. Vartabedian VF, Savage PB, Peyton L (2016) The processing and presentation of lipids and glycolipids to the immune system. Immunol Rev 272:109–119 - DOI
    1. Ishikawa E, Ishikawa T, Morita YS et al (2009) Direct recognition of the mycobacterial glycolipid, trehalose dimycolate, by C-type lectin Mincle. J Exp Med 206:2879–2888 - DOI

Publication types

LinkOut - more resources