Hypoallergenic variants of the Parietaria judaica major allergen Par j 1: A member of the non-specific lipid transfer protein plant family

Background. Par j 1 represents a major allergenic component of Parietaria j udaica (Pj) pollen, since it is able to induce an immunoglobulin E (IgE) re sponse in 95% of Pj-allergic patients. It belongs to the non-specific lipid transfer protein family, sharing with them a common three-dimensional stru cture. Methods: Disulphide bond variants of the recombinant Par j 1 (rParj 1) allergen were generated by site-directed mutagenesis, and the immunologi cal activity of rPar j 1 and its conformational mutants was compared with t he use of the skin prick test (SPT). The ability to bind IgE antibodies was evaluated by Western blot, ELISA and ELISA inhibition. T cell reactivity w as measured by peripheral blood mononuclear cell proliferation assay. Resul ts: The disruption of Cys14-Cys29 and Cys30-Cys75 bridging (PjA mutant) cau sed the loss of the majority of specific IgE-binding activity. Additional d isruption of the Cys4-Cys52 bridge (PjC mutant) and the latter Cys50-Cys91 bridge (PjD mutant) led to the abolition of IgE-binding activity. On the SP T, PjB (lacking the Cys4-Cys52 and Cys50-Cys91 bridges) was still capable o f triggering a type 1 hypersensitive reaction in 9 out of 10 patients, and PjA in 3 out of 10 patients, while PjC and PjD did not show any SPT reactiv ity. All the mutants preserved their T cell reactivity. Conclusion: Recombi nant hypoallergenic variants of the rPar j 1 allergen described herein may represent a useful tool for improved immunotherapy. Copyright (C) 2001 S, K arger AG, Basel.