title: Time- and concentration-dependent effects of exogenous serotonin and inflammatory cytokines on mast cell function.
abstract: Mast cells play a significant role in both the innate and adaptive immune response; however, the tissue-bound nature of mast cells presents an experimental roadblock to performing physiologically relevant mast cell experiments. In this work, a heterogeneous cell culture containing primary culture murine peritoneal mast cells (MPMCs) was studied to characterize the time-dependence of mast cell response to allergen stimulation and the time- and concentration-dependence of the ability of the heterogeneous MPMC culture to uptake and degranulate exogenous serotonin using high performance liquid chromatography (HPLC) coupled to an electrochemical detector. Additionally, because mast cells play a central role in asthma, MPMCs were exposed to CXCL10 and CCL5, two important asthma-related inflammatory cytokines that have recently been shown to induce mast cell degranulation. MPMC response to both allergen exposure and cytokine exposure was evaluated for 5-HT secretion and bioactive lipid formation using ultraperformance liquid chromatography coupled to an electrospray ionization triple quadrupole mass spectrometer (UPLC-MS/MS). In this work, MPMC response was shown to be highly regulated and responsive to subtle alterations in a complex environment through time- and concentration-dependent degranulation and bioactive lipid formation. These results highlight the importance of selecting an appropriate mast cell model when studying mast cell involvement in allergic response and inflammation.
authors.affiliation: Department of Chemistry, University of Minnesota , 207 Pleasant St. SE, Minneapolis, Minnesota 55455, United States.
authors.forename: Sarah M
authors.initials: SM
authors.lastname: Gruba
authors.forename: Audrey F
authors.initials: AF
authors.lastname: Meyer
authors.forename: Benjamin M
authors.initials: BM
authors.lastname: Manning
authors.forename: Yiwen
authors.initials: Y
authors.lastname: Wang
authors.forename: John W
authors.initials: JW
authors.lastname: Thompson
authors.forename: Joseph J
authors.initials: JJ
authors.lastname: Dalluge
authors.forename: Christy L
authors.initials: CL
authors.lastname: Haynes
mesh_terms: D000818:Animals; D015550:Cell Degranulation; D002478:Cells, Cultured; D018946:Chemokine CCL5 / Q000276:immunology; D054357:Chemokine CXCL10 / Q000276:immunology; D016207:Cytokines / Q000276:immunology*; D007073:Immunoglobulin E / Q000276:immunology; D007249:Inflammation / Q000276:immunology; D008407:Mast Cells / Q000166:cytology / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D012701:Serotonin / Q000276:immunology*; D053719:Tandem Mass Spectrometry
chemical_list: D018946:Chemokine CCL5; D054357:Chemokine CXCL10; D016207:Cytokines; D012701:Serotonin; D007073:Immunoglobulin E