Effects of phosphatidylinositol-3-kinase inhibitors on degranulation and gene induction in allergically triggered mouse mast cells.
BACKGROUND
Phosphatidylinositol-3-kinase (PI3-kinase) comprises an essential component in a number of signaling cascades, primarily of the growth factor type. Two specific inhibitors, wortmannin and demethoxyviridin (DMV), are widely used to block signaling via this molecule and link certain receptors to the PI3-kinase pathway.


METHODS
We have studied the extent of involvement of PI3-kinase in signaling events by Fc epsilonRI in mast cells using a mouse mast cell line as a model system. This was done using beta-hexosaminidase release assays, a leukotriene ELISA, transient transfections with reporter gene constructs of TNF alpha and MARC, and in addition a TNF alpha ELISA.


RESULTS
Consistent with previously published data in the rat basophilic cell line RBL-2H3, we find that wortmannin as well as DMV prevent the degranulation reaction in the mouse mast cell line CPII. DMV also inhibits the release of leukotrienes, leading to the conclusion that Fc epsilonRI activates PI3-kinase which then mediates these reactions. On the contrary, however, lymphokine and chemokine induction at the gene and protein level is not inhibited, suggesting that the activation of this gene set in mast cells is independent of PI3-kinase.


CONCLUSIONS
PI3-kinase is activated in our mast cell model system via cross-linking of the Fc epsilonRI. This reaction is clearly necessary for the degranulation process and the release of leukotrienes. Activation of lymphokine and chemokine genes as well as secretion of their gene products are not triggered along the PI3-kinase signaling pathway. This is in agreement with our previous findings, showing that the MAP kinase pathway and Ca2+ influx are both involved in gene activation in this cell type.
Pendl
G G
GG
Sandoz Research Institute, Department of Immunodermatology, Vienna, Austria.
Prieschl
E E
EE
Thumb
W
W
Harrer
N E
NE
Auer
M
M
Baumruker
T
T
D000485:Allergens / Q000276:immunology*; D000730:Androstadienes / Q000494:pharmacology; D000736:Androstenes / Q000494:pharmacology; D000818:Animals; D015550:Cell Degranulation / Q000187:drug effects* / Q000276:immunology; D002460:Cell Line; D004136:Dinitrobenzenes / Q000276:immunology; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000276:immunology; D007073:Immunoglobulin E / Q000494:pharmacology; D020024:Leukotriene Antagonists; D015289:Leukotrienes / Q000378:metabolism; D008407:Mast Cells / Q000201:enzymology* / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D019869:Phosphatidylinositol 3-Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000037:antagonists & inhibitors*; D015533:Transcriptional Activation; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D000077191:Wortmannin
D000485:Allergens; D000730:Androstadienes; D000736:Androstenes; D004136:Dinitrobenzenes; D020024:Leukotriene Antagonists; D015289:Leukotrienes; D014409:Tumor Necrosis Factor-alpha; D007073:Immunoglobulin E; C070594:demethoxyviridin; D019869:Phosphatidylinositol 3-Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor); D000077191:Wortmannin