Pierre Massion
Faculty Member
Last active: 1/11/2018

Fluorescence-based measurement of cystine uptake through xCT shows requirement for ROS detoxification in activated lymphocytes.

Siska PJ, Kim B, Ji X, Hoeksema MD, Massion PP, Beckermann KE, Wu J, Chi JT, Hong J, Rathmell JC
J Immunol Methods. 2016 438: 51-58

PMID: 27594594 · PMCID: PMC5065394 · DOI:10.1016/j.jim.2016.08.013

T and B lymphocytes undergo metabolic re-programming upon activation that is essential to allow bioenergetics, cell survival, and intermediates for cell proliferation and function. To support changes in the activity of signaling pathways and to provide sufficient and necessary intracellular metabolites, uptake of extracellular nutrients increases sharply with metabolic re-programming. One result of increased metabolic activity can be reactive oxygen species (ROS), which can be toxic when accumulated in excess. Uptake of cystine allows accumulation of cysteine that is necessary for glutathione synthesis and ROS detoxification. Cystine uptake is required for T cell activation and function but measurements based on radioactive labeling do not allow analysis on single cell level. Here we show the critical role for cystine uptake in T cells using a method for measurement of cystine uptake using a novel CystineFITC probe. T cell receptor stimulation lead to upregulation of the cystine transporter xCT (SLC7a11) and increased cystine uptake in CD4+ and CD8+ human T cells. Similarly, lipopolysaccharide stimulation increased cystine uptake in human B cells. The CystineFITC probe was not toxic and could be metabolized to prevent cystine starvation induced cell death. Furthermore, blockade of xCT or competition with natural cystine decreased uptake of CystineFITC. CystineFITC is thus a versatile tool that allows measurement of cystine uptake on single cell level and shows the critical role for cystine uptake for T cell ROS regulation and activation.

Copyright © 2016 Elsevier B.V. All rights reserved.

MeSH Terms (18)

Amino Acid Transport System y+ B-Lymphocytes Cell Line, Tumor Cellular Reprogramming Cystine Flow Cytometry Fluorescein-5-isothiocyanate Fluorescence Fluorescent Dyes Glutathione Humans Lymphocyte Activation Microscopy, Fluorescence Reactive Oxygen Species Receptors, Antigen, T-Cell Signal Transduction T-Lymphocytes Up-Regulation

Connections (1)

This publication is referenced by other Labnodes entities: