IMMUNO-METABOLISM LINKAgilent 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.

IFNγControl

LPS+IFNγLPS

Time (min)

Basal

Lymph

Neutro

PltMono

0 120 240 360 4800

500

1000

1500

PER

(pm

ol/m

in)

Time (min)

Oligomycin

Injection (or Control)

FCCPControl orEtomoxir

Rotenone +Antimycin A

0 25 50 75 100 125 1500

200

400

600

800

1000

OCR

(pM

oles

/min

)

ECAR (mpH/min/µg protein)0 1 2 3 4 5 6 7 8 9 10

0123456789

10

OCR

(pm

oles

/min

/µg

prot

ein)

+Oligo

IFNγControl

LPS+IFNγLPS

Time (min)

Basal

Lymph

Neutro

PltMono

0 120 240 360 4800

500

1000

1500

PER

(pm

ol/m

in)

Time (min)

Oligomycin

Injection (or Control)

FCCPControl orEtomoxir

Rotenone +Antimycin A

0 25 50 75 100 125 1500

200

400

600

800

1000

OCR

(pM

oles

/min

)

ECAR (mpH/min/µg protein)0 1 2 3 4 5 6 7 8 9 10

0123456789

10

OCR

(pm

oles

/min

/µg

prot

ein)

+Oligo

IFNγControl

LPS+IFNγLPS

Time (min)

Basal

Lymph

Neutro

PltMono

0 120 240 360 4800

500

1000

1500

PER

(pm

ol/m

in)

Time (min)

Oligomycin

Injection (or Control)

FCCPControl orEtomoxir

Rotenone +Antimycin A

0 25 50 75 100 125 1500

200

400

600

800

1000

OCR

(pM

oles

/min

)

ECAR (mpH/min/µg protein)0 1 2 3 4 5 6 7 8 9 10

0123456789

10

OCR

(pm

oles

/min

/µg

prot

ein)

+Oligo

XFe 96Kramer et al., (2014) Redox Biol

XF Assays & Kits provide routine, reliable, and consistent results for:

• Mitochondrial Function• Glycolytic Function• Metabolic Phenotype• Cell Fuel Flexibility/Preference

XF Technology to measure:

• 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

More about Immunologyhttp://www.agilent.com/en/solutions/cell-metabolism-(seahorse)/immunology-research

Buy onlinewww.chem.agilent.com/store

Publications Databasewww.agilent.com/publications-database/

Find a local Agilent customer center in your countrywww.agilent.com/chem/contactus

Worldwide Technical Supportseahorse.support@agilent.com

USA and Canada1-800-227-9770; select option 3 then 8

EuropeUK: 0800 096 7632 Denmark: +45 8830 5083 Germany: 0800 180 66 78 Netherlands: 0800 022 7243 Other EU: +45 3136 9878

Assay Kits Available:XF Real Time Activation AssayXF Cell Mito Stress Test KitXF Glycolysis Stress Test Kit

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

This information is subject to change without notice.

© Agilent Technologies, Inc. 2017 Printed in the USA October 17, 2017 5991-8618EN

Researchers are using XF Technology to Push the Boundaries of Immunology Research

Learn More About Seahorse XF Technology and Immunology Research agilent.com/chem/immunology

XFe 96

A comprehensive window into the entire immune cell life cycle

Cell Lineage and Metabolism

NaiveT cell

ActivateT cell

EffectorT cell

MemoryT cell

Fatty acid oxidation

Glycolysis (ECAR)

Mitochondrial respiration (OCR)

XF Glycolytic Rate Assay KitXF Mito Fuel Flex Test KitXF Cell Energy Phenotype Test Kit

Adapted from Hongbo Chi, (2012) Nat Rev Immunol