Engineered recombinant ovomucoid third domain can modulate allergenic response in Balb/c mice model

• Published in: Biochemical and biophysical research communications 2006-04, Vol.342 (3), p.710-717
• Main Authors: Rupa, Prithy, Mine, Yoshinori
• Format: Article
• Language: English
• Subjects: Allergens - chemistry; Allergens - immunology; Animals; Antigens - isolation & purification; Balb/c mice; Cytokines - metabolism; Disease Models, Animal; Female; Food allergy; Hypersensitivity - immunology; IgE; Immunoglobulin E - blood; Immunoglobulin E - metabolism; Immunoglobulin G - blood; Immunoglobulin G - metabolism; Immunotherapy; Interferon-gamma - metabolism; Interleukin-12 - metabolism; Interleukin-4 - metabolism; Mice; Mice, Inbred BALB C; Ovomucin - chemistry; Ovomucin - immunology; Ovomucin - isolation & purification; Ovomucoid; Protein Structure, Tertiary; Recombinant Proteins - chemistry; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; Site-directed mutagenesis; Spleen - cytology; T-Lymphocytes - immunology; Th1/Th2
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2006.01.174

IgE-mediated allergic reactions to egg white are a serious health problem and ovomucoid being the dominant egg white allergen has been on focus in the past decade. Engineered hypoallergens with reduced reactivity for IgE antibodies are being examined to modulate the allergic response and develop prophylactic allergen vaccines. In this study, we evaluated the immunomodulatory effect of a genetic variant of the third domain of ovomucoid (GMFA) which showed reduced IgE binding with egg allergic patient’s sera in comparison to the native form of the third domain of ovomucoid (DIII) in a murine model system. Balb/c mice were injected intraperitoneally with DIII and GMFA antigens. Allergen-specific serum IgG, IgG1, IgG2a, and IgE responses were evaluated using enzyme-linked immunosorbent assay. Splenocyte cytokine levels in the medium of the cultured cells were examined by ELISA and levels of IL-4, INF-γ, and IL-12 (p70) cytokines were quantified. Neutralization with anti-IL-12 monoclonal antibody was assayed and cytokine levels with respect to GMFA mutant antigen stimulation were measured. GMFA mutant form was found to have significantly reduced levels of specific IgE when compared to the DIII suggesting a mutation-induced abrogation of the IgE binding epitope in mice. The increase in IgG2a levels in GMFA together with the decline of IgE and IgG1 points to a shift from a Th2 response to a Th1 dominated response. The cytokine profile showed a modulation of anti-allergic Th1 phenotype in GMFA from a proallergic Th2 response observed with DIII. Low levels of IL-4 and increased levels of INF-γ and IL-12 were observed and anti-IL-12 monoclonal antibody restored the levels of IL-4 and suppressed levels of INF-γ and IL-12 in the GMFA sensitized group. These results indicate that GMFA has a marked suppressive effect on the allergic response of ovomucoid and caused a shift towards a Th1 pathway, thereby modulating the Th1/Th2 cytokine balance and could be used as a potential hypoallergenic candidate for allergen-immunotherapy in the treatment of egg white allergy.