Subthreshold desensitization of human basophils re-capitulates the loss of Syk and FcεRI expression characterized by other methods of desensitization.
BACKGROUND
Clinical desensitization of patients to drugs involves progressive exposure to escalating doses of drug over a period of 24 h. In prior studies, this method was re-capitulated in vitro to also demonstrate loss of mast cell or basophil responsiveness. However, most signalling studies of human basophils have identified changes in signalling by using other methods of inducing cellular desensitization.


OBJECTIVE
This study examined two well-described endpoints of basophil desensitization, loss of syk or FcεRI expression, under conditions of subthreshold desensitization.


METHODS
The loss of FcεRI and syk was examined in human basophils.


RESULTS
It was shown that both loss of syk and FcεRI/IgE occurred during an escalating series of stimulation (anti-IgE Ab) and that expression loss occurred despite the presence of little histamine release. If basophils were first cultured for 3 days in 10 ng/mL IL-3, the concentration-dependence of histamine release shifted to 100-fold lower concentrations of stimulus. However, loss of syk did not show any change in its EC50 while loss of FcεRI also shifted 100-fold. From the perspective of early signal element activation, the marked shift in the EC50 for histamine release was not accompanied by similar shifts in the EC50s for several signalling elements. The EC50s for phospho-Src, phospho-SHIP1, phospho-Syk, or phospho-Cbl did not change while the EC50s for phospho-Erk and the cytosolic calcium response did shift 100-fold.


CONCLUSIONS
These studies show that under normal conditions, subthreshold desensitization leads to loss of two critical signalling molecules (FcεRI and syk) but under at least one condition, treatment with IL-3, it is possible to markedly blunt the loss of syk, but not FcεRI, while executing a proper subthreshold titration. These data also suggest that IL-3 modifies only the sensitivity of signalling elements that are downstream of syk activation.
Johns Hopkins Asthma and Allergy Center, Baltimore, MD 21224, USA. dmacglas@jhmi.edu
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D000906:Antibodies / Q000494:pharmacology; D001491:Basophils / Q000276:immunology / Q000378:metabolism*; D002478:Cells, Cultured; D003888:Desensitization, Immunologic; D004342:Drug Hypersensitivity / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D005786:Gene Expression Regulation; D006632:Histamine / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology; D047908:Intracellular Signaling Peptides and Proteins / Q000276:immunology; D011505:Protein-Tyrosine Kinases / Q000096:biosynthesis* / Q000276:immunology; D017455:Receptors, IgE / Q000096:biosynthesis* / Q000276:immunology; D015398:Signal Transduction / Q000276:immunology; D000072377:Syk Kinase
D000906:Antibodies; C508596:IL3 protein, human; D007377:Interleukin-3; D047908:Intracellular Signaling Peptides and Proteins; D017455:Receptors, IgE; D006632:Histamine; D011505:Protein-Tyrosine Kinases; C000606896:SYK protein, human; D000072377:Syk Kinase