Activation-secretion coupling in 10P2 murine mast cells challenged with IgE-antigen, ionophore A23187, thapsigargin and phorbol ester.
Mast cell activation-secretion by several signal transduction pathways results in the release of proinflammatory mediators including histamine, proteases, arachidonic acid metabolites and multifunctional cytokines. In the present investigations the activation-secretion responses of the cytokine-independent, cloned 10P2 cell line have been explored. [14C]Serotonin (5-HT) preloaded cells were stimulated with antigen, with and without IL-4, ionophore A23187, thapsigargin or phorbol myristate acetate (PMA). Following passive sensitization with anti-dinitrophenol (anti-DNP) IgE, mast cells released up to 31% of incorporated [14C]5-HT when stimulated with specific antigen (DNP-human serum albumin). This response was potentiated by pretreatment with IL-4. Significant degranulation (50%) was noted following treatment with calcium ionophore A23187, thapsigargin and ionophore A23187/PMA. Collectively, these results suggest that 10P2 cells undergo activation-secretion responses, assessed as degranulation of preloaded [14C]5-HT when challenged with IgE antigen, by influx of extracellular calcium or release of intracellular calcium stores, or by direct activation of protein kinase isozymes. As a growth factor-independent cell line, 10P2 cells may be a valuable adjunct to existing mast cell model systems currently used for pharmacologic investigations.
Heiman
A S
AS
College of Pharmacy and Pharmaceutical Sciences, Florida A & M University, Tallahassee 32307, USA.
Chen
M
M
D019297:2,4-Dinitrophenol / Q000378:metabolism; D000704:Analysis of Variance; D000818:Animals; D000941:Antigens / Q000633:toxicity*; D000001:Calcimycin / Q000633:toxicity*; D002273:Carcinogens / Q000633:toxicity*; D002460:Cell Line; D007116:Immunization, Passive; D007073:Immunoglobulin E / Q000276:immunology; D015847:Interleukin-4 / Q000633:toxicity*; D007476:Ionophores / Q000633:toxicity*; D008407:Mast Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D012701:Serotonin / Q000378:metabolism / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects; D013755:Tetradecanoylphorbol Acetate / Q000633:toxicity*; D019284:Thapsigargin / Q000633:toxicity*
D000941:Antigens; D002273:Carcinogens; D007476:Ionophores; D015847:Interleukin-4; D012701:Serotonin; D007073:Immunoglobulin E; D000001:Calcimycin; D019284:Thapsigargin; D013755:Tetradecanoylphorbol Acetate; D019297:2,4-Dinitrophenol