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Updated: May 2, 2026
An Ex vivo Mast Cell Degranulation Assay using Crude Peritoneal Exudate Cells and Natural Antigen Stimulation
Published on: April 27, 2021
Inhibiting mast cell degranulation by HO-1 affects dendritic cell maturation in vitro
Yuan-yuan Ma1, Mu-qing Yang, Chun-feng Wang
1Department of General Surgery, Shanghai Tenth People's Hospital of Tong Ji University, Shanghai, China.
Objective And Design:
Mast cell (MC) degranulation can break peripheral immune tolerance. However, its mechanism remains unclear. Our goal was to study the stabilization of MC membranes by heme oxygenase-1 (HO-1) in order to influence dendritic cell (DC) function.
Material:
Mast cells and dendritic cells were prepared from 8-week-old to 10-week-old C57BL/6 mice; spleen mononuclear cells (SMCs) were prepared from 8-week-old to 10-week-old C57BL/6 and Balb/c mice.
Treatment:
Mast cells were pretreated with PBS, DMSO, Hemin (50 μl/ml), and Znpp (50 μl/ml) for 8 h.
Method:
Real-time PCR and western-blot tested the HO-1 of MC mRNA and protein. The co-stimulatory molecules of DCs (CD80, CD86, CD40) were measured by flow cytometry, and levels of TNF-α, IL-6, and IFN-γ were measured by ELISA. We set up a one-way mixed lymphocyte reaction (MLR) model to test the proliferation of SMCs after MC/DC interaction. *P < 0.05 (t test) was taken as the level of statistical significance.
Result:
MCs pretreated with hemin induced HO-1 mRNA and protein expression, then interacted with DCs; expression of the co-stimulatory molecules was attenuated. The TNF-α, IL-6, and IFN-γ levels in the co-culture system were decreased. These DCs couldn't stimulate the proliferation of SMCs.
Conclusion:
Inhibiting MC degranulation by HO-1 restrained DC maturation and attenuated the proliferation of SMCs.
