Background: FcεRIβ reportedly functions as an amplifier of the FcεRIγ-mediated activation signal using a reconstitution system. However, the amplification mechanisms in human mast cells (MCs) are poorly understood. We previously reported the hyperexpression of FcεRIβ of MCs in giant papillae from vernal keratoconjunctivitis patients, compared with that in conjunctivae from nonallergic conjunctivitis patients. Elucidation of the molecular mechanisms of the amplification induced by FcεRIβ should provide new targets for novel therapeutic interventions. The aim is to understand in greater details the function of FcεRIβ in human MC FcεRI expression and signaling.
Methods: FcεRIβ and Lyn expression was reduced using a lentiviral shRNA silencing technique. Localization of Lyn and FcεRIβ in cultured MCs was examined by confocal microscopic analysis. Mediators were measured by ELISAs.
Results: The diminution of FcεRIβ significantly downregulated cell surface FcεRI expression and FcεRI-mediated mediator release/production. The downregulation of FcεRI-mediated degranulation was not only due to the decrease in FcεRI expression. The diminution of FcεRIβ inhibited the redistribution of Lyn within the cell membrane following IgE sensitization. The diminution of Lyn in MCs significantly downregulated FcεRI-mediated degranulation. The recombinant cell-penetrating forms of phosphorylated FcεRIβ immunoreceptor tyrosine-based activation motif (ITAM) for intracellular delivery disturbed the interaction between Lyn and phosphorylated endogenous FcεRIβ ITAM, resulted in inhibiting IgE-dependent histamine release from MCs in vitro and from giant papillae specimens ex vivo.
Conclusion: The interaction between Lyn and FcεRIβ is indispensable for FcεRI-mediated human MC activation, and specific inhibition of the interaction may represent a new therapeutic strategy for the treatment of human allergic diseases.
© 2012 John Wiley & Sons A/S.