Abstract
The small molecule 8-methoxy-2-oxo-1,2,4,5-tetrahydrocyclopenta[de]quinoline-3-carboxylic acid (2b) behaves as a reactive non-fluorescent Michael acceptor, which after reaction with thiols becomes fluorescent, and an efficient Eu3+ antenna, after self-assembling with this cation in water. This behavior makes 2b a highly selective GSH biosensor, which has demonstrated high potential for studies in murine and human cells of the immune system (CD4+ T, CD8+ T, and B cells) using flow cytometry. GSH can be monitored by the fluorescence of the product of addition to 2b (445 nm) or by the luminescence of Eu3+ (592 nm). 2b was able to capture baseline differences in GSH intracellular levels among murine and human CD4+ T, CD8+ T, and B cells. We also successfully used 2b to monitor intracellular changes in GSH associated with the metabolic variations governing the induction of CD4+ naïve T cells into regulatory T cells (TREG).
Highlights
2b a highly selective GSH biosensor, which has demonstrated high potential for studies in murine and human cells of the immune system (CD4+ T, CD8+ T, and B cells) using flow cytometry
Several studies have shed light on the dynamic and sophisticated connection between metabolic programs and the function of specialized cells in the immune system.[10,11]. This crucial role of metabolism in the control of immune processes, including inflammation, has led to the emergence of a new field of immunometabolism.[11−13] It is increasingly recognized that biothiols play a key role in regulating the metabolic adaptability and thereby the function of cells of the immune system.[14−21] One of the latest discoveries in this field is the regulation of functions through the synthesis and release of various biothiols, in particular, GSH, which affects the metabolism and function of the immune system’s effector cells.[12,17,21−24] the interest in developing tools to monitor biothiol levels in immune cells in clinical samples has grown exponentially
Article high luminescence lifetime and narrow emission bands, which allow an increase in the sensitivity and signal-to-noise ratio, avoiding natural background fluorescence in time-resolved luminescence spectroscopy.[33−35] Among the few lanthanidebased biothiol sensors reported in the literature,[36,37] to our knowledge, no lanthanide antenna-based sensors, which selfassemble in water have yet been reported
Summary
2b a highly selective GSH biosensor, which has demonstrated high potential for studies in murine and human cells of the immune system (CD4+ T, CD8+ T, and B cells) using flow cytometry.
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