Analysis of glycogen metabolic pathway utilization by dendritic cells and T cells using custom phenotype metabolic assays

J Immunol Methods. 2018 Jul:458:53-57. doi: 10.1016/j.jim.2018.04.006. Epub 2018 Apr 22.

Abstract

In the field of immunology, there is an increasing interest in cellular energy metabolism and its outcome on immune cell effector function. Activation of immune cells leads to rapid metabolic changes that are central to cellular biology in order to support the effector responses. Therefore, the need for user-friendly and dependable assay technologies to address metabolic regulation and nutrient utilization in immune cells is an important need in this field. Redox-dye reduction-based Phenotype MicroArray (PM) assays, which measure NADH reduction as a readout, developed by Biolog Inc., provide a wide screening of metabolites both in bacteria and mammalian cells. In this study, we delineate a detailed protocol of a customized Biolog assay for investigation of a specific metabolic pathway of interest. The option to be able to easily customize this technology offers researchers with a convenient assay platform to methodically examine specific nutrient substrates or metabolic pathways of interest in a rapid and cost effective manner.

Keywords: Biolog; Dendritic cells; Glucose; Glycogen; Metabolism; Phenotypic MicroArray.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amides / pharmacology
  • Animals
  • Biological Assay / instrumentation
  • Biological Assay / methods*
  • Cell Culture Techniques
  • Cell Line
  • Coloring Agents / chemistry
  • Dendritic Cells / drug effects
  • Dendritic Cells / immunology
  • Dendritic Cells / metabolism
  • Energy Metabolism / drug effects
  • Energy Metabolism / immunology
  • Glycogen / analysis
  • Glycogen / metabolism
  • Glycogen Phosphorylase / antagonists & inhibitors
  • Glycogen Phosphorylase / metabolism
  • Humans
  • Indoles / pharmacology
  • Metabolic Networks and Pathways / drug effects
  • Metabolic Networks and Pathways / immunology
  • Mice
  • Microarray Analysis / instrumentation
  • Microarray Analysis / methods*
  • Oxidation-Reduction
  • T-Lymphocytes / drug effects
  • T-Lymphocytes / immunology
  • T-Lymphocytes / metabolism

Substances

  • Amides
  • CP 91149
  • Coloring Agents
  • Indoles
  • Glycogen
  • Glycogen Phosphorylase