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Updated: Jul 21, 2025
Isolation of Leukocytes from the Human Maternal-fetal Interface
Published on: May 21, 2015
Decidual lymphatic endothelial cell-derived granulocyte-macrophage colony-stimulating factor induces M1 macrophage
Yun Ji Jung1, Yeji Lee1, Hayan Kwon1
1Department of Obstetrics and Gynecology, Institute of Women's Medical Life Science, Placenta-derived Stem Cell and Genomic Research Lab, Yonsei University College of Medicine, Yonsei University Health System, Seoul, The Republic of Korea.
Problem:
Direct interactions between macrophages and lymphatic vessels have been shown previously. In pre-eclampsia (PE), macrophages are dominantly polarized into a proinflammatory M1 phenotype and lymphangiogenesis is defective in the decidua. Here, we investigated whether decidual lymphatic endothelial cells (dLECs) affect macrophage polarization in PE.
Method Of Study:
THP-1 macrophages were cocultured with dLECs or cultured in the conditioned medium (CM) of dLECs. Macrophage polarization was measured using flow cytometry. Granulocyte-macrophage colony-stimulating factor (GM-CSF) expression in dLECs was measured using qRT-PCR and ELISA. The activation of nuclear translocation of nuclear factor-κ (NF-κB), an upstream signaling molecule of GM-CSF, was assessed by immunocytochemical localization of p65. Through GM-CSF knockdown and NF-κB inhibition in dLEC, we evaluated whether the GM-CSF/NF-κB pathway of PE dLEC affects decidual macrophage polarization.
Results:
The ratio of inflammatory M1 macrophages with HLA-DR+ /CD80+ markers significantly increased following coculturing with PE dLECs or culturing in PE dLEC CM, indicating that the PE dLEC-derived soluble factor acts in a paracrine manner. GM-CSF expression was significantly upregulated in PE dLECs. Recombinant human GM-CSF induced macrophage polarization toward an M1-like phenotype, whereas its knockdown in PE dLECs suppressed it, suggesting PE dLECs induce M1 macrophage polarization by secreting GM-CSF. The NF-κB p65 significantly increased in PE dLECs compared to the control, and pretreatment with an NF-κB inhibitor significantly suppressed GM-CSF production from PE dLECs.
Conclusions:
In PE, dLECs expressing high levels of GM-CSF via the NF-κB-dependent pathway play a role in inducing decidual M1 macrophage polarization.
Insights
In pre-eclampsia, decidual lymphatic endothelial cells (dLECs) promote inflammatory M1 macrophage polarization via granulocyte-macrophage colony-stimulating factor (GM-CSF) signaling, driven by the NF-κB pathway.
Area of Science:
- Reproductive immunology
- Vascular biology
- Maternal-fetal medicine
Background:
- Direct interactions between macrophages and lymphatic vessels are established.
- Pre-eclampsia (PE) is characterized by M1 macrophage polarization and defective decidual lymphangiogenesis.
- Decidual lymphatic endothelial cells (dLECs) role in macrophage polarization in PE is unclear.
Purpose of the Study:
- To investigate the effect of decidual lymphatic endothelial cells (dLECs) on macrophage polarization in pre-eclampsia (PE).
- To elucidate the underlying molecular mechanisms involving GM-CSF and NF-κB signaling.
Main Methods:
- Co-culture of THP-1 macrophages with PE dLECs or their conditioned medium (CM).
- Flow cytometry for macrophage polarization assessment.
- qRT-PCR, ELISA, and immunocytochemistry for GM-CSF and NF-κB (p65) analysis.
- GM-CSF knockdown and NF-κB inhibition in dLECs.
Main Results:
- PE dLECs or their CM significantly increased M1 macrophage polarization, indicating paracrine signaling.
- Granulocyte-macrophage colony-stimulating factor (GM-CSF) expression was upregulated in PE dLECs.
- GM-CSF induced M1 polarization; GM-CSF knockdown in dLECs suppressed it.
- NF-κB activation (p65 nuclear translocation) was increased in PE dLECs and inhibition reduced GM-CSF production.
Conclusions:
- Decidual lymphatic endothelial cells (dLECs) in pre-eclampsia (PE) induce M1 macrophage polarization.
- This effect is mediated by granulocyte-macrophage colony-stimulating factor (GM-CSF) secretion.
- The NF-κB-dependent pathway in dLECs drives GM-CSF production and subsequent M1 macrophage polarization in PE.
