In vitro inhibition of human conjunctival mast-cell degranulation by ketotifen.
Ketotifen relieves the symptoms of allergic conjunctivitis through multiple mechanisms of action. One such mechanism may involve stabilization of conjunctival mast cells. Because of inter- and intra-species variation, however, this hypothesis cannot be adequately tested using mast cells from animals or other human tissues. We therefore employed human conjunctival mast cells. The mast cells were prepared using human conjunctival tissues obtained from US eye banks. Cell suspensions were sensitized with human IgE and incubated with ketotifen fumarate or control. After antigenic challenge of sensitized cells with anti-IgE, levels of histamine and tryptase, two mast-cell granule markers, were measured in the supernatant fluid. Cell viability was assessed with a Trypan Blue assay. Ketotifen at concentrations of approximately 10(-11) to 10(-4) M inhibited mast-cell histamine release by 90% or more. Similarly, ketotifen at approximately 10(-10) to 10(-4) M inhibited tryptase release by 90% or more (apart from a single anomalous reading). At all ketotifen concentrations that stabilized mast cells, cell viability was preserved. Moreover, ketotifen did not impair cell viability unless concentrations were increased above the clinically relevant range, i.e., above the order of magnitude of 10(-4) M. These data demonstrate that ketotifen can stabilize human conjunctival mast cells, without impairing cell viability.
Schoch
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Novartis Ophthalmics AG, Basel, Switzerland. christian.schoch@pharma.novartis.com
D018926:Anti-Allergic Agents / Q000008:administration & dosage / Q000494:pharmacology*; D015550:Cell Degranulation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D003228:Conjunctiva / Q000166:cytology*; D004305:Dose-Response Relationship, Drug; D006634:Histamine H1 Antagonists / Q000008:administration & dosage / Q000494:pharmacology*; D006636:Histamine Release / Q000187:drug effects; D006801:Humans; D066298:In Vitro Techniques; D007665:Ketotifen / Q000008:administration & dosage / Q000494:pharmacology*; D008407:Mast Cells / Q000378:metabolism / Q000502:physiology*; D009994:Osmolar Concentration; D012697:Serine Endopeptidases / Q000378:metabolism; D053802:Tryptases
D018926:Anti-Allergic Agents; D006634:Histamine H1 Antagonists; D012697:Serine Endopeptidases; D053802:Tryptases; D007665:Ketotifen