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Macrophage Activation State Markers

Click on one of the macrophage activation states shown in the buttons below to see the markers that are commonly used to identify each activated state or view a list of the common macrophage markers used for distinguishing macrophages from other immune cell types.

Property title
M1 Macrophage
M1 Macrophage
Stimuli
Stimuli
IFN-gamma
IFN-gamma
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Secreted Factors
Secreted Factors
CCL2/MCP-1
CCL2/MCP-1
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CCL3/MIP-1 alpha
CCL3/MIP-1 alpha
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CCL4/MIP-1 beta
CCL4/MIP-1 beta
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CCL5/RANTES
CCL5/RANTES
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CCL8/MCP-2
CCL8/MCP-2
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CCL11/Eotaxin
CCL11/Eotaxin
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CCL15/MIP-1 delta (human)
CCL15/MIP-1 delta (human)
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CCL19/MIP-3 beta
CCL19/MIP-3 beta
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CCL20/MIP-3 alpha
CCL20/MIP-3 alpha
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CXCL1/KC
CXCL1/KC
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CXCL2/MIP-2
CXCL2/MIP-2
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CXCL3/GRO gamma
CXCL3/GRO gamma
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CXCL5/ENA-78
CXCL5/ENA-78
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CXCL8/IL-8
CXCL8/IL-8
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CXCL9/MIG
CXCL9/MIG
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CXCL10/IP-10
CXCL10/IP-10
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CXCL11/I-TAC
CXCL11/I-TAC
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CXCL13/BLC/BCA-1
CXCL13/BLC/BCA-1
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CXCL16
CXCL16
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CX3CL1/Fractalkine
CX3CL1/Fractalkine
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IL-1 beta/IL-1F2
IL-1 beta/IL-1F2
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IL-6
IL-6
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IL-12 high
IL-12 high
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IL-15
IL-15
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IL-17
IL-17
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IL-18
IL-18
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IL-23
IL-23
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IFN-gamma
IFN-gamma
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TNF-alpha
TNF-alpha
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Cell Surface Markers
Cell Surface Markers
B7-1/CD80+
B7-1/CD80+
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B7-2/CD86+
B7-2/CD86+
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CD36/SR-B3+
CD36/SR-B3+
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CD68/SR-D1+
CD68/SR-D1+
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CD163-
CD163-
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Fc gamma RII/CD32+
Fc gamma RII/CD32+
Fc gamma RIII/CD16+
Fc gamma RIII/CD16+
HLA-DR high (human)
HLA-DR high (human)
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IFN-gamma R+
IFN-gamma R+
Steady-State Macrophage
Steady-State Macrophage
MHC class II high
MHC class II high
Intracellular
Markers
Intracellular
Markers
COX-2+
COX-2+
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iNOS+
iNOS+
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IRF5+
IRF5+
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STAT1+
STAT1+
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Lipopolysaccharide
Lipopolysaccharide
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OxLDL
OxLDL
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Cholesterol crystals
Cholesterol crystals
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Overview

M1 Macrophage Activation State Markers Overview

Macrophages are activated by exogenous stimuli in their environments such as cytokines, growth factors, or microbial agonists, which can alter their transcriptional programs and functional phenotypes. The historical model of macrophage activation describes two different macrophage activation states, classical and alternative macrophage activation. Classical M1 macrophage activation is stimulated by pro-inflammatory cytokines and microbial products including IFN-gamma and/or lipopolysaccharide or TNF-alpha. The resulting M1 macrophages secrete pro-inflammatory mediators such as TNF-alpha, IL-1 beta, IL-12, IL-18, IL-23, nitric oxide, and reactive oxygen species, which have anti-microbial and tumoricidal properties and drive Th1- and Th17-mediated immune responses. Common cell surface markers used to identify M1 macrophages include high levels of MHC class II, CD68, B7-1/CD80, B7-2/CD86, and iNOS.

Data Examples

Data Examples

Detection of Cell Surface Markers on Resting and M1 or M2a Activated Human Macrophages by Flow Cytometry. Enriched human CD14+ monocytes were cultured in the presence of 50 ng/mL Recombinant Human GM-CSF (R&D Systems, Catalog # 215-GM) or 50 ng/mL Recombinant Human M-CSF (R&D Systems, Catalog # 216-MC) for 6 days in a RPMI-based media containing 10% FBS. Following the 6-day maturation period, macrophages were activated for an additional 24 hours to either the classical M1 macrophage phenotype with 50 ng/mL lipopolysaccharide and 50 ng/mL Recombinant Human IFN-gamma (R&D Systems, Catalog # 285-IF; solid blue histograms) or alternatively activated M2a macrophage phenotype with 20 ng/mL Recombinant Human IL-4 and 20 ng/mL Recombinant Human IL-13 (R&D Systems, Catalog # 204-IL and Catalog # 213-ILB, respectively; solid purple histograms). Macrophages that were not activated were cultured for an additional 24 hours in media containing either GM-CSF (light blue histograms) or M-CSF (light purple histograms) so that all macrophages were cultured for a total of 7 days before being compared by flow cytometry. Cells were then stained with either a fluorochrome-conjugated Mouse Anti-Human B7-1/CD80 Monoclonal Antibody (R&D Systems, Catalog # FAB140), a fluorochrome-conjugated Mouse Anti-Human B7-2/CD86 Monoclonal Antibody (R&D Systems, Catalog # FAB141), or a fluorochrome-conjugated Mouse Anti-Human CD163 Monoclonal Antibody (R&D Systems, Catalog # FAB1607). Isotype control staining is indicated by the solid gray histograms. Zombie Violet cell viability dye labeling and doublet exclusion were used to remove dead cells from the surface marker expression analysis.