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Updated: Jul 15, 2026
Isolation of Murine Peritoneal Macrophages to Carry Out Gene Expression Analysis Upon Toll-like Receptors Stimulation
Published on: April 29, 2015
Human peritoneal macrophages show functional characteristics of M-CSF-driven anti-inflammatory type 2 macrophages
Wei Xu1, Nicole Schlagwein, Anja Roos
1Department of Nephrology, Leiden University Medical Center, Leiden, The Netherlands.
Abstract:
We have recently shown that in vitro polarized M-CSF-driven anti-inflammatory macrophages (MPhi2) have the unique capacity to preferentially bind and ingest early apoptotic cells. However, these data are based on in vitro polarized cells and it is unclear whether MPhi2-like cells exist in vivo. Here we used CD163 as a cell surface marker to distinguish MPhi2 from the pro-inflammatory MPhi1. We show that human peritoneal MPhi (pMPhi) freshly isolated from patients on peritoneal dialysis have the phenotypical characteristics of MPhi2, including CD163 surface expression and lack of CD16. Like MPhi2, pMPhi have the capacity for endocytosis and macropinocytosis, are able to preferentially bind and ingest early apoptotic cells, and produce large amounts of IL-10 upon stimulation with LPS. Moreover, upon LPS stimulation both pMPhi and MPhi2 down-regulate CD86, resulting in a reduced capacity to stimulate proliferation of allogeneic T cells and an inhibition of Th1 cytokine release of these T cells. Our data provide the evidence for the first time that in vitro polarized MPhi2 exist in vivo, and human pMPhi resemble the anti-inflammatory MPhi2. We propose that pMPhi have the potential to maintain an anti-inflammatory condition in the peritoneal cavity.
Insights
Human peritoneal macrophages (pMPhi) resemble in vitro-generated anti-inflammatory macrophages (MPhi2). These pMPhi cells can clear apoptotic cells and maintain an anti-inflammatory environment in the peritoneal cavity.
Area of Science:
- Immunology
- Cell Biology
Background:
- In vitro studies demonstrated that M-CSF-driven anti-inflammatory macrophages (MPhi2) can bind and ingest early apoptotic cells.
- It remained unclear if MPhi2-like cells exist in vivo.
Purpose of the Study:
- To investigate the in vivo existence and characteristics of MPhi2-like cells.
- To determine if human peritoneal macrophages (pMPhi) resemble MPhi2.
Main Methods:
- Used CD163 as a cell surface marker to differentiate MPhi2 from MPhi1.
- Analyzed phenotypical characteristics, endocytosis, macropinocytosis, apoptotic cell binding/ingestion, and cytokine production (IL-10) of freshly isolated human pMPhi.
- Assessed the impact of LPS stimulation on CD86 expression and T cell proliferation/cytokine release.
Main Results:
- Freshly isolated human pMPhi exhibited MPhi2 characteristics, including CD163 expression and lack of CD16.
- pMPhi demonstrated capacities for endocytosis, macropinocytosis, and preferential binding/ingestion of early apoptotic cells.
- LPS stimulation of pMPhi and MPhi2 led to CD86 downregulation, reduced T cell proliferation, and inhibited Th1 cytokine release.
Conclusions:
- This study provides the first evidence that in vitro-polarized MPhi2 exist in vivo.
- Human pMPhi closely resemble anti-inflammatory MPhi2.
- pMPhi possess the potential to establish and maintain an anti-inflammatory state within the peritoneal cavity.
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