Inducing an anergic state in mast cells and basophils without secretion.
BACKGROUND
IgE-mediated secretion from mast cells or basophils depends on the activity of both spleen tyrosine kinase (syk) and phosphatidyl inositol 3' kinase (PI3K), but several specific downregulatory pathways (eg, loss of syk expression) do not.


OBJECTIVE
We tested whether stimulation with antigen in the presence of a syk inhibitor (NVP-QAB205) would ablate secretion while simultaneously allowing anergy.


METHODS
The anergic or desensitized state in human basophils, cultured-derived mast cells, and in situ stimulated airway mast cells (in organ baths) was assessed after stimulation with antigen in the presence of syk inhibitor.


RESULTS
Antigen caused 35 +/- 7% and 62 +/- 10% histamine release from basophils and mast cells, respectively, and it caused an 87 +/- 5% histamine/leukotriene D(4)-dependent contraction of human isolated bronchi. All of these responses were blocked >95% by the syk inhibitor. Rechallenging the preparations with antigen, after first washing out the syk inhibitor and antigen, revealed that near complete anergy (92% to 100%) occurred in each case. A similar result was found when using a PI3K inhibitor, LY294002, in studies of basophils.


CONCLUSIONS
Although the syk inhibitor nearly abolished the antigen-induced secretion from mast cells and basophils, it had little effect on the pathways involved in anergy. These results suggest that syk and PI3K are not involved in downregulation leading to anergy.
Asthma and Allergy Center, Johns Hopkins University, Baltimore, MD 21224, USA. dmacglas@jhmi.edu
Donald
D
MacGlashan
Bradley J
BJ
Undem
D001491:Basophils / Q000187:drug effects / Q000502:physiology*; D015550:Cell Degranulation / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D017635:Clonal Anergy / Q000502:physiology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006636:Histamine Release; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000037:antagonists & inhibitors; D008407:Mast Cells / Q000187:drug effects / Q000502:physiology*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D000072377:Syk Kinase
D004791:Enzyme Inhibitors; D047908:Intracellular Signaling Peptides and Proteins; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011505:Protein-Tyrosine Kinases; C000606896:SYK protein, human; D000072377:Syk Kinase