title: Generation of a chimeric dust mite hypoallergen using DNA shuffling for application in allergen-specific immunotherapy.
abstract: Specific immunotherapy (SIT) is the only treatment that provides long lasting relief of allergy symptoms. Unfortunately, SIT-based traditional remedies carry the risk of producing local and/or systemic side effects. To improve the safety and efficacy of SIT, it has been proposed that SIT must utilize allergens that are hypoallergenic but hyperimmunogenic. Therefore, we used DNA shuffling to generate mutant genes encoding hypoallergens with potent immunogenicity and screened them for their capacity to modify the allergic response. We tentatively shuffled the major group 1 allergen genes from house dust mite, Dermatophagoides farinae and Dermatophagoides pteronyssinus, and discovered a novel chimeric gene, termed C 1. The gene was expressed in Escherichia coli (E. coli) and the chimeric protein C 1 was purified. An animal model of asthma demonstrated that C 1 not only decreased the production of serum IgE and IgG1, and inhibited the production of IL-4 and IL-5 in the bronchoalveolar lavage fluid (BALF). C 1 also boosted the levels of IgG2a and IFN-γ, which may demonstrate a rebalance of TH1 and TH2 allergic response. Additionally, flow cytometry showed that the immunogenicity of C 1 was higher than that of ProDer f 1, but was not significantly different from that of ProDer p 1. Our findings suggest that the C 1 is hypoallergenic and yet highly immunogenic, which makes it potentially safe and effective for use in SIT of allergic asthma.
authors.affiliation: Department of Medical Parasitology, School of Basic Medicine, Wannan Medical College Wuhu 241002, Anhui, China.
authors.forename: Bei-Bei
authors.initials: BB
authors.lastname: Zhao
authors.affiliation: Department of Medical Parasitology, School of Basic Medicine, Wannan Medical College Wuhu 241002, Anhui, China.
authors.forename: Ji-Dong
authors.initials: JD
authors.lastname: Diao
authors.affiliation: Department of Medical Parasitology, School of Basic Medicine, Wannan Medical College Wuhu 241002, Anhui, China.
authors.forename: Zhi-Ming
authors.initials: ZM
authors.lastname: Liu
authors.affiliation: Department of Medical Parasitology, School of Basic Medicine, Wannan Medical College Wuhu 241002, Anhui, China.
authors.forename: Chao-Pin
authors.initials: CP
authors.lastname: Li
authors.affiliation: Department of Physiology, School of Basic Medicine, Wannan Medical College Wuhu 241002, Anhui, China.
authors.forename: Yu-Xin
authors.initials: YX
authors.lastname: Jiang
mesh_terms: D000818:Animals; D039741:Antigens, Dermatophagoides / Q000235:genetics* / Q000276:immunology; D001249:Asthma / Q000517:prevention & control; D015153:Blotting, Western; D032621:DNA Shuffling / Q000379:methods*; D003888:Desensitization, Immunologic / Q000379:methods*; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology* / Q000517:prevention & control; D008807:Mice, Inbred BALB C; D039981:Pyroglyphidae; D018183:Transplantation Chimera
chemical_list: D039741:Antigens, Dermatophagoides
keywords: DNA shuffling; Dermatophagoides allergen 1 group; allergenicity; immunogenicity; specific immunotherapy