title: Mast cells cultured from IL-3-treated mice show impaired responses to bacterial antigen stimulation.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  Mast cells were obtained from the peritoneal cavity of C57BL/6 mice.
  
  
  METHODS
  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.
  
  
  CONCLUSIONS
  Mast cells cultured from IL-3-treated mice show impaired responses.
authors: Parasite Immune Modulation Group, School of Nursing and Human Sciences, Dublin City University, Glasnevin, Dublin, Ireland.
authors: Krisztina V
authors: KV
authors: Vukman
authors: Tamás
authors: T
authors: Visnovitz
authors: Paul N
authors: PN
authors: Adams
authors: Martin
authors: M
authors: Metz
authors: Marcus
authors: M
authors: Maurer
authors: Sandra M
authors: SM
authors: O'Neill
mesh_terms: D000818:Animals; D000942:Antigens, Bacterial / Q000378:metabolism*; D001886:Bordetella pertussis / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D007249:Inflammation; D007377:Interleukin-3 / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000737:chemistry; D008407:Mast Cells / Q000166:cytology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018482:Muscle, Skeletal / Q000378:metabolism; D010537:Peritoneum / Q000166:cytology / Q000378:metabolism; D013154:Spleen / Q000378:metabolism; D013997:Time Factors
chemical_list: D000942:Antigens, Bacterial; D007377:Interleukin-3; D008070:Lipopolysaccharides