IMMUNO-METABOLISM LINK Agilent Seahorse XF Technology Technical Flyer Move beyond characterization to controlling cell fate by modulating metabolism • Activate • Modulate • Investigate Immune cells have distinct metabolic phenotypes Seahorse XF Technology • Live cell • Real-time • Label-free • Dynamic injection ports • Measures both major metabolic pathways simultaneously – respiration and glycolysis Platelets and leukocytes have distinct metabolic signatures Adapted from Kramer et al., (2014) Redox Biol Macrophage activation increases glycolytic rates (LPS and INFγ) Yoon et al., (2017) 5991-8342EN Fatty acid oxidation is required for memory T cell development Adapted from Van der Windt et al., (2012) Immunity XF Technology Provides a Window into Immune Cell Function Simultaneously measure major metabolic pathways that regulate immune cell function Both measurements work together as one simple indicator for richer, and more complete information. XF Rapid Real Time cell analysis to discriminate activation and cell function Time-resolved information enables a better understanding of dynamic mechanisms and continued response. XF Compound injection to investigate: Fuels, Signaling, and Pathways Injection ports deliver up to 4 compounds, in addition to immunogenic activation beads. Basal Lymph Neutro Plt Mono ECAR (mpH/min/μg protein) 0 1 2 3 4 5 6 7 8 9 10 0 1 2 3 4 5 6 7 8 9 10 OCR (pmoles/min/μg protein) +Oligo IFNγ Control LPS+IFNγ LPS Time (min) 0 120 240 360 480 0 500 1000 1500 PER (pmol/min) Injection (or Control) Time (min) Oligomycin FCCP Control or Etomoxir Rotenone + Antimycin A 0 25 50 75 100 125 150 0 200 400 600 800 1000 OCR (pMoles/min) XFe 96 Kramer et al., (2014) Redox Biol
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IMMUNO-METABOLISM LINKAdapted from Van der Windt et al., (2012) Immunity XF Technology Provides a Window into Immune Cell Function Simultaneously measure major metabolic pathways that
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• Immune cell life cycle, lineage, and checkpoints• Immune cell phenotype, function, and reprogramming• Real time activation • Metabolic requirements and fuels• Metabolic signaling and pathways• Modulation to control outcome
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