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Updated: May 27, 2026
Analyzing the Functions of Mast Cells In Vivo Using 'Mast Cell Knock-in' Mice
Published on: May 27, 2015
Mast cells cultured from IL-3-treated mice show impaired responses to bacterial antigen stimulation
Krisztina V Vukman1, Tamás Visnovitz, Paul N Adams
1Parasite Immune Modulation Group, School of Nursing and Human Sciences, Dublin City University, Glasnevin, Dublin, Ireland.
Objective And Design:
This study exploits the biological activity of interleukin (IL)-3 to generate high yields of peritoneal mast cells ex vivo in order to examine pro-inflammatory immune responses in ex-vivo culture.
Material Or Subjects:
Mast cells were obtained from the peritoneal cavity of C57BL/6 mice.
Treatment:
Mice were injected intraperitoneally twice per day for 5 days with IL-3 (40-50 μg/ml) to increase mast cell numbers.
Methods:
Histological studies examined mast cell numbers in the peritoneal cavity, intestine, lung, spleen and skeletal muscle. Peritoneal mast cells cultured ex vivo (PCMCs) were stimulated for 24 h with lipopolysaccharide and Bordetella pertussis antigen and secretion of tumour necrosis factor-α, IL-6, IL-4, IL-5, IL-10 and interferon-γ into supernatant was measured by commercial ELISA. Cell surface marker expression of FcεRI, c-kit, OX40L and TLR2 was measured by flow cytometry. Mast cell degranulation was measured using a β-hexosaminidase assay.
Results:
IL-3 treatment increases mast cell numbers in the peritoneal cavity, spleen and muscle but not intestine and lung of C57BL/6 mice. PCMCs generated from IL-3-treated mice exhibit impaired growth, differentiation and responses to activation as measured by decreased cytokine secretion and cell surface marker expression.
Conclusion:
Mast cells cultured from IL-3-treated mice show impaired responses.
