Published in:
01-01-2012 | Original Research Paper
Mast cells cultured from IL-3-treated mice show impaired responses to bacterial antigen stimulation
Published in: Inflammation Research | Issue 1/2012
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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.