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. [C-14]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 [C-14]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 [C-14]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.