Characterization of an immunomodulatory Der p 2-FIP-fve fusion protein produced in transformed rice suspension cell culture

• Published in: Transgenic research 2012-02, Vol.21 (1), p.177-192
• Main Authors: Su, Chin-Fen, Kuo, I-Chun, Chen, Peng-Wen, Huang, Chiung-Hui, Seow, See Voon, Chua, Kaw Yan, Yu, Su-May
• Format: Article
• Language: English
• Subjects: Adjuvants; Allergenicity; Allergens; Allergic diseases; alpha -Amylase; alpha-Amylases - genetics; Animal Genetics and Genomics; Animals; antibodies; Antigens, Dermatophagoides - genetics; Antigens, Dermatophagoides - immunology; Arthropod Proteins - genetics; Arthropod Proteins - immunology; Basidiocarps; Biological and medical sciences; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Biotechnology; Blood - drug effects; Blood - metabolism; Cell culture; Cell Culture Techniques - methods; cell suspension culture; culture media; Culture Media - metabolism; Cytokines; Cytokines - metabolism; Dermatophagoides pteronyssinus; Enzyme-Linked Immunosorbent Assay; Flammulina velutipes; Fundamental and applied biological sciences. Psychology; Fungal Proteins - genetics; Fungal Proteins - immunology; Fusion protein; genes; Genetic Engineering; Genetic technics; Helper cells; Hemagglutination; Hemagglutination Tests; Histamine; Histamine - metabolism; Humans; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Immunologic Factors - genetics; Immunologic Factors - metabolism; Immunomodulation; Immunotherapy; Life Sciences; Lymphocytes; Lymphocytes T; Methods. Procedures. Technologies; Mice; Mice, Inbred C57BL; mites; Molecular Medicine; mushrooms; Original Paper; Oryza - cytology; Oryza - genetics; Oryza sativa; patients; Plant Genetics and Genomics; Plants, Genetically Modified; Promoter Regions, Genetic; Promoters; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - immunology; Recombinant Fusion Proteins - metabolism; Recombinant Fusion Proteins - pharmacology; rice; Starvation; Sucrose; Th2 Cells - immunology; Th2 Cells - metabolism; Transgenic animals and transgenic plants; Transgenics
• ISSN: 0962-8819; 1573-9368
• DOI: 10.1007/s11248-011-9518-6

Der p 2, a major allergen of Dermatophagoides pteronyssinus mites, is one of the most clinically relevant allergens to allergic patients worldwide. FIP-fve protein (Fve) from the golden needle mushroom (Flammulina velutipes) is an immunomodulatory protein with potential Th1-skewed adjuvant properties. Here, we produced and immunologically evaluated a Der p 2-Fve fusion protein as a potential immunotherapeutic for allergic diseases. Using an inducible expression system in cultured rice suspension cells, the recombinant Der p 2-Fve fusion protein (designated as OsDp2Fve) was expressed in rice cells under the control of an α-amylase gene (αAmy8) promoter and secreted under sucrose starvation. OsDp2Fve was partially purified from the cultured medium. The conformation of Der p 2 in OsDp2Fve remains intact as reflected by its unaltered allergenicity, as assessed by human IgE ELISA and histamine release assays, compared to non-fusion Der p 2 protein. Furthermore, the Fve protein expressed in OsDp2Fve retains its in vitro lymphoproliferative activity but loses its hemagglutination and lymphoagglutination effects compared to the native protein. Notably, in vivo evaluation showed that mice administered with OsDp2Fve possessed an enhanced production of Der p 2-specific IgG antibodies without potentiating the production of Der p 2-specific IgE and Th2 effector cytokines in comparison with mice co-administered with native Fve and Der p 2 proteins. These results suggest that the recombinant Der p 2-Fve fusion protein produced in rice suspension cell cultures has a great potential for allergy immunotherapy.