Cellular and Molecular Life Sciences

, Volume 68, Issue 18, pp 3095–3107 | Cite as

Novel macrophage polarization model: from gene expression to identification of new anti-inflammatory molecules

  • Gloria Lopez-Castejón
  • Alberto Baroja-Mazo
  • Pablo Pelegrín
Research Article


Plasticity is a well-known property of macrophages that is controlled by different changes in environmental signals. Macrophage polarization is regarded as a spectrum of activation phenotypes adjusted from one activation extreme, the classic (M1), to the other, the alternative (M2) activation. Here we show, in vitro and in vivo, that both M1 and M2 macrophage phenotypes are tightly coupled to specific patterns of gene expression. Novel M2-associated markers were characterized and identified as genes controlling the extracellular metabolism of ATP to generate pyrophosphates (PPi). Stimulation of M1 macrophages with PPi dampens both NLR and TLR signaling and thus mediates cytokine production. In this context extracellular PPi enhanced the resolution phase of a murine peritonitis model via a decrease in pro-inflammatory cytokine production. Therefore, our study reveals an additional level of plasticity modulating the resolution of inflammation.


Alternatively activated macrophages Resolution of inflammation Toll-like receptors Nucleotide-binding domain and leucin-rich repeat receptors Extracellular pyrophosphates Ectonucleotidases Cytokines Peritonitis 



Progressive ankylosis disease susceptibility gene product


Apoptotic speck-like protein with a caspase-activating recruiting domain


Ectonucleotide pyrophosphatase/phosphodiesterase


Lactate dehydrogenase


Classically activated macrophages


Alternatively activated macrophages


Nucleotide-binding domain and leucin-rich repeat receptors






Reactive oxygen species


Toll-like receptors

Supplementary material

18_2010_609_MOESM1_ESM.tif (1.2 mb)
Supplemental Fig. 1. Expression of different housekeeping genes during macrophage polarization gradient. A Real-time quantitative (qRT)-PCR for the indicated house-keeping genes (GAPDH, YWHAZ, HPRT1, and SDHA) during the polarization gradient protocol. B qRT-PCR for TNF-α gene expression in M1, M1/M2, or M2 polarized macrophages normalized to the four different housekeeping genes. No significant differences were found in the fold increase/decrease trend among the different macrophage phenotypes. (TIFF 1259 kb)
18_2010_609_MOESM2_ESM.tif (1.4 mb)
Supplemental Fig. 2. Expression of the 39 analyzed genes in extreme M1 or M2 macrophages. Real-time quantitative RT-PCR for the indicated genes in extreme M1 (A) or M2 (B) polarization. M2 genes are represented in green, M1 in red, and housekeeping genes in blue. Relative expression normalized to YWHAZ was log2 transformed. (TIFF 1398 kb)
18_2010_609_MOESM3_ESM.tif (2.1 mb)
Supplemental Fig. 3. Gene expression in extreme M1 or M2 macrophages. Dot plot representing log2 transformed expression data from real-time quantitative RT-PCR normalized values for all genes included in this study (Suppl. Fig. 2). Data for each individual gene are represented in accordance with its expression in extreme M1 or M2 genes. R2 = 0.341. (TIFF 2157 kb)


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Copyright information

© Springer Basel AG 2010

Authors and Affiliations

  • Gloria Lopez-Castejón
    • 2
  • Alberto Baroja-Mazo
    • 1
  • Pablo Pelegrín
    • 1
    • 2
  1. 1.Inflammation and Experimental Surgery UnitUniversity Hospital “Virgen de la Arrixaca”-Fundación Formación Investigación Sanitaria Región Murcia (FFIS)MurciaSpain
  2. 2.Faculty of Life ScienceUniversity of ManchesterManchesterUK

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