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The Agilent Advantage - Measure T Cell Activation in Minutes

Cellular energy metabolism is now understood to influence many immune cell pathways as both an indicator and controller of immune cell function and lineage commitment. This concept is clearly illustrated in the activation of naïve T cells to stimulate proliferation.
Activate
  • Measure CD4+ & CD8+ T cell activation within 6 minutes of stimulation with CD3/CD28 beads.
Modulate
  • Introduce up to 3 modulators per well with programmable drug injection ports to elucidate T cell response to activation kinetics.
Investigate
  • Explore the metabolism driving
    T cell response through assays providing metabolic phenotype, bioenergetic pathway, and fuel/substrate preference.
This Application Note describes a method for quantifying T cell activation using in situ activation of naïve T cells with immunogenic beads injected during the assay. This method is rapid, kinetic, nondestructive, and directly tied to cellular function.
Adapted from Hongbo Chi, Nat Rev Immunol.;
12(5): 325-338, 2012
The Cell Lineage and Immunometabolism Link
Advances in the field of immunology show cellular metabolism as a driver and regulator of immune cell function and differentiation. T cell activation is correlated with metabolic shifts toward glycolysis, activating energy promoting pathways required for high rates of cellular proliferation.
Accelerate your analysis from hours to minutes -
a new method to assess T cell activation
Based on recent discoveries, glycolytic pathway dependency for energy production is a robust measure of T cell activation. Conventionally, progression of T cell activation is measured in terms of changes in cell size/morphology, interleukin/interferon expression and cell surface markers, methods which can be time and labor intensive, often involve end-point (not kinetic) data, and a time scale of hours to days. The Seahorse XFp analyzer can detect T cell activation response within several minutes of stimulation with CD3/CD28 immunogenic beads by measuring glycolytic extracellular acidification rate (ECAR) in real time. Learn more about XF technology.
Modulate T cell activation
with the use of programmable drug injection ports
Agilent Seahorse XF sensor cartridges incorporate integrated injection ports that sequentially deliver up to 3 compounds, in addition to immunogenic activation beads, allowing dose response, agonist or antagonist response, or pathway perturbation analysis of each sample.
Dive deeper with standardized cell metabolism assays and powerful software
In addition to T cell activation, Seahorse metabolic assay kits are available for measuring mitochondrial respiration, glycolysis, and fatty acid oxidation in cells. Seahorse XF kits and reagents are pre-calibrated, pre-tested and are coupled with powerful report generator software, providing functional metabolic data from live T cells.
Metabolic Phenotype
Rapidly determines the energy phenotype/fingerprint of cells and reveals their metabolic potential
Bioenergetic Pathways
Provides a complete mitochondrial and glycolytic profile of your T cells using mitochondrial and glycolysis stress test kits
Fuels Preference
Measures the dependency, capacity, and flexibility of cells to oxidize any one of three mitochondrial fuels: glucose, glutamine, or fatty acids
This tutorial describes a novel assay that measures immune cell activation from within minutes of activation throughout the fully developed activated phenotype in both a qualitative and quantitative manner. This provides a sensitive window of opportunity to identify and characterize attributes that can drive immune responses to desired outcomes.



For Research Use Only. Not for use in diagnostic procedures.