Analysis of sphingosine kinase activity in single natural killer cells from peripheral blood

Integrative Biology : Quantitative Biosciences From Nano to Macro
Alexandra J DickinsonNancy L Allbritton

Abstract

Sphingosine-1-phosphate (S1P), a lipid second messenger formed upon phosphorylation of sphingosine by sphingosine kinase (SK), plays a crucial role in natural killer (NK) cell proliferation, migration, and cytotoxicity. Dysregulation of the S1P pathway has been linked to a number of immune system disorders and therapeutic manipulation of the pathway has been proposed as a method of disease intervention. However, peripheral blood NK cells, as identified by surface markers (CD56(+)CD45(+)CD3(-)CD16) consist of a highly diverse population with distinct phenotypes and functions and it is unknown whether the S1P pathway is similarly diverse across peripheral blood NK cells. In this work, we measured the phosphorylation of sphingosine-fluorescein (SF) and subsequent metabolism of S1P fluorescein (S1PF) to form hexadecanoic acid fluorescein (HAF) in 111 single NK cells obtained from the peripheral blood of four healthy human subjects. The percentage of SF converted to S1PF or HAF was highly variable amongst the cells ranging from 0% to 100% (S1PF) and 0% to 97% (HAF). Subpopulations of cells with varying levels of S1PF formation and metabolism were readily identified. Across all subjects, the average percentage of SF converted to S1PF...Continue Reading

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Citations

Aug 22, 2015·Expert Review of Gastroenterology & Hepatology·He Joe WangMary Katherine Jung
Feb 24, 2016·Biomedicine & Pharmacotherapy = Biomédecine & Pharmacothérapie·Fangdong GuoLin Zhang
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Feb 19, 2016·Electrophoresis·Damith E W PatabadigeChristopher T Culbertson
Jan 30, 2018·ACS Chemical Biology·Brianna M VickermanDavid S Lawrence

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