title: Stem cell factor and interleukin-4 increase responsiveness of mast cells to substance P.
abstract: The response of mast cells (MC) to non-IgE-mediated stimulation is critically dependent on the population of MC examined. The neuropeptide Substance P (SP) has been reported to activate connective tissue-type MC (CTMC), while mucosal MC (MMC) are not activated by SP. We examined the effect of stem cell factor (SCF) plus interleukin-4 (IL-4) on SP-initiated activation of bone marrow-derived MC (BMMC). Mouse MC, derived from a culture of BM cells with IL-3, were subsequently treated with recombinant SCF plus IL-4 for 6 days. Responsiveness to SP was monitored measuring beta-hexosaminidase and lipid mediator release. Histochemical staining, histamine analysis, and granule protease expression were achieved to characterize the cells. In contrast to IL-3 grown cells, SCF/IL-4-exposed cells showed functional responsiveness to release beta-hexosaminidase (42.25% +/- 1.46% at SP concentration of 100 microM) and produce leukotriene C(4) (LTC(4)) (7.4 +/- 1.5 ng/10(6) cells)/prostaglandin D(2) (PGD(2)) (2.0 +/- 0.3 ng/10(6) cells) upon stimulation by SP. The increase in sensitivity of the cells to SP was not due to differentiation into CTMC, as the cells remained heparin negative. Both SCF and IL-4 were needed because SCF or IL-4 alone were insufficient to keep cells viable after 3 to 4 days post coculture. SP-induced secretion from BMMC cultured in medium containing SCF plus IL-4 (25.76% +/- 1.83%) was higher in comparison with cells cultured with SCF plus IL-3 (8.85% +/- 0.68%).These findings indicate that temporal changes in cytokine expression can influence the sensitivity of MC to non-immunologic stimuli. Local cytokine production leading to an increase in MC responsiveness to SP and inducing secretion of granule content and lipid generation may, therefore, propagate and worsen inflammatory conditions.
authors.affiliation: Department of Pharmacology and Pathophysiology, Utrecht Institute for Pharmaceutical Sciences, Faculty of Pharmacy, Utrecht University, Utrecht, The Netherlands.
authors.forename: K
authors.initials: K
authors.lastname: Karimi
authors.forename: F A
authors.initials: FA
authors.lastname: Redegeld
authors.forename: R
authors.initials: R
authors.lastname: Blom
authors.forename: F P
authors.initials: FP
authors.lastname: Nijkamp
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000187:drug effects; D002478:Cells, Cultured; D003594:Cytoplasmic Granules / Q000378:metabolism; D004357:Drug Synergism; D006632:Histamine / Q000096:biosynthesis; D007377:Interleukin-3 / Q000494:pharmacology; D015847:Interleukin-4 / Q000494:pharmacology*; D017997:Leukotriene C4 / Q000096:biosynthesis*; D008407:Mast Cells / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008817:Mice, Mutant Strains; D018945:Monocyte Chemoattractant Proteins / Q000096:biosynthesis; D015230:Prostaglandin D2 / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019089:Stem Cell Factor / Q000494:pharmacology*; D013373:Substance P / Q000494:pharmacology*; D001619:beta-N-Acetylhexosaminidases / Q000096:biosynthesis
chemical_list: C099677:CCL13 protein, human; D007377:Interleukin-3; D018945:Monocyte Chemoattractant Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D019089:Stem Cell Factor; D015847:Interleukin-4; D017997:Leukotriene C4; D013373:Substance P; D006632:Histamine; D001619:beta-N-Acetylhexosaminidases; D015230:Prostaglandin D2