A Simple Flow Cytometric Method to Measure Glucose Uptake and Glucose Transporter Expression for Monocyte Subpopulations in Whole Blood

Abstract

Monocytes are innate immune cells that can be activated by pathogens and inflammation associated with certain chronic inflammatory diseases. Activation of monocytes induces effector functions and a concomitant shift from oxidative to glycolytic metabolism that is accompanied by increased glucose transporter expression. This increased glycolytic metabolism is also observed for trained immunity of monocytes, a form of innate immunological memory. Although in vitro protocols examining glucose transporter expression and glucose uptake by monocytes have been described, none have been examined by multi-parametric flow cytometry in whole blood. We describe a multi-parametric flow cytometric protocol for the measurement of fluorescent glucose analog 2-NBDG uptake in whole blood by total monocytes and the classical (CD14 ++ CD16 -), intermediate (CD14 ++ CD16 + ) and non-classical (CD14 + CD16 ++) monocyte subpopulations. This method can be used to examine glucose transporter expression and glucose uptake for total monocytes and monocyte subpopulations during homeostasis and inflammatory disease, and can be easily modified to examine glucose uptake for other leukocytes and leukocyte subpopulations within blood.

Human immunodeficiency virus and antiretroviral therapy-mediated immune cell metabolic dysregulation in children born to HIV-infected women: potential clinical implications

“…A study by Mason and colleagues demonstrated that GLUT-1 expression positively correlated with mitochondrial mass and mitochondrial membrane potential ( 70 ). Another study in adult PLWHA demonstrated that HIV infection caused an increase in the uptake of a fluorescent glucose analogue, 2NBDG, by infected monocytes ( 71 ). Remarkably, upregulated glucose metabolism has been associated with CD4+ T lymphocytes apoptosis although it is unknown whether these changes may be involved in the decrease of CD4+ lymphocytes in HIV infection ( 72 , 73 ).…”

Non-Enzymatic Glucose Sensors Composed of Polyaniline Nanofibers with High Electrochemical Performance

“…There is also a great demand for glucose monitors in the food and chemical industry. Researchers have developed many methods of detection, including flow cyclometric, liquid chromatography, mass spectroscopy, and fluorescence [ 5 , 6 , 7 , 8 ]. However, these methods have limitations, such as low accuracy, high cost, and complexity of operation.…”