Plasma membrane lipid rafts are highly ordered membrane microdomains enriched for glycosphingolipids and cholesterol, which play an important role during T-cell antigen receptor (TCR) signaling. Our previous work has demonstrated that plasma membrane lipid composition is an important determinant of human CD4+ T-cell function and that defects in lipid raft expression contribute to CD4+ dysfunction in patients with autoimmunity. In this chapter we share three flow cytometry-based methods to quantitatively analyze plasma membrane lipid composition in primary human CD4+ T cells. We describe the quantification of glycosphingolipid expression using cholera toxin subunit B, cholesterol expression using filipin staining, and membrane “lipid order” using di-4-ANEPPDHQ. These methods can easily be adapted to analyze different cell types.
CITATION STYLE
Waddington, K. E., Pineda-Torra, I., & Jury, E. C. (2019). Analyzing T-cell plasma membrane lipids by flow cytometry. In Methods in Molecular Biology (Vol. 1951, pp. 209–216). Humana Press Inc. https://doi.org/10.1007/978-1-4939-9130-3_16
Mendeley helps you to discover research relevant for your work.