Abstract
Design of hypoallergen with low IgE reactivity is desirable for allergen-specific immunotherapy. Despite oyster tropomyosin (Cra g 1) is considered as the major allergen, no immunotherapy is available now. In the current research, we generated hypoallergens of Cra g 1 and evaluated their allergenicity. Four hypoallergenic derivatives were constructed by epitope deletion or site-directed mutagenesis on grounds of the identified epitopes. They showed obvious reduction in reactivity towards IgE from oyster-allergic patients and Cra g 1-sensitized BN rats, as well as significant decrease in degranulation and secretion of allergic mediators including histamine, IL-4, IL-6 and TNF-α. In addition, to further investigate the molecular mechanism, we examined the effects of these variants on FcεRI-dependent signalling pathway in IgE-challenged RBL-2H3 cells. We found that the hypoallergenic mutants were able to attenuate FcεRI-mediated signaling cascades in tested cells. These results indicate that the hypoallergenic molecules have ideal characteristics and offer a promising new strategy in clinical immunotherapy for shellfish-allergic subjects.
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