title: 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.
authors: Centre for Biomedical Research, Macfarlane Burnet Institute for Medical Research and Public Health; Department of Infectious Diseases, Monash University; Department of Microbiology and Immunology, University of Melbourne; cpalmer@burnet.edu.au.
authors: Clovis S
authors: CS
authors: Palmer
authors: Department of Microbiology, The University of the West Indies.
authors: Joshua J
authors: JJ
authors: Anzinger
authors: Department of Microbiology, The University of the West Indies.
authors: Tiffany R
authors: TR
authors: Butterfield
authors: Division of Experimental Medicine, Department of Medicine, University of California, San Francisco.
authors: Joseph M
authors: JM
authors: McCune
authors: Centre for Biomedical Research, Macfarlane Burnet Institute for Medical Research and Public Health; Department of Infectious Diseases, Monash University; Department of Medicine, Monash University.
authors: Suzanne M
authors: SM
authors: Crowe
mesh_terms: D005434:Flow Cytometry / Q000379:methods*; D005947:Glucose; D051246:Glucose Transport Proteins, Facilitative / Q000378:metabolism*; D006801:Humans; D018950:Lipopolysaccharide Receptors; D009000:Monocytes / Q000276:immunology / Q000378:metabolism*; D017452:Receptors, IgG / Q000378:metabolism
chemical_list: D051246:Glucose Transport Proteins, Facilitative; D018950:Lipopolysaccharide Receptors; D017452:Receptors, IgG; D005947:Glucose