title: Characterization of an immunomodulatory Der p 2-FIP-fve fusion protein produced in transformed rice suspension cell culture.
abstract: 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.
authors.affiliation: Institute of Biotechnology, National Cheng Kung University, No.1, University Road, Tainan 701, Taiwan, ROC.
authors.forename: Chin-Fen
authors.initials: CF
authors.lastname: Su
authors.forename: I-Chun
authors.initials: IC
authors.lastname: Kuo
authors.forename: Peng-Wen
authors.initials: PW
authors.lastname: Chen
authors.forename: Chiung-Hui
authors.initials: CH
authors.lastname: Huang
authors.forename: See Voon
authors.initials: SV
authors.lastname: Seow
authors.forename: Kaw Yan
authors.initials: KY
authors.lastname: Chua
authors.forename: Su-May
authors.initials: SM
authors.lastname: Yu
mesh_terms: D000818:Animals; D039741:Antigens, Dermatophagoides / Q000235:genetics* / Q000276:immunology; D060829:Arthropod Proteins / Q000235:genetics* / Q000276:immunology; D001769:Blood / Q000187:drug effects / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D003470:Culture Media / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005656:Fungal Proteins / Q000235:genetics* / Q000276:immunology; D006386:Hemagglutination Tests; D006632:Histamine / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G; D007155:Immunologic Factors / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D012275:Oryza / Q000166:cytology / Q000235:genetics*; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology; D018418:Th2 Cells / Q000276:immunology / Q000378:metabolism; D000516:alpha-Amylases / Q000235:genetics
chemical_list: D039741:Antigens, Dermatophagoides; D060829:Arthropod Proteins; D003470:Culture Media; D016207:Cytokines; C451163:Dermatophagoides pteronyssinus antigen p 2; C547132:FIP-fve protein, Flammulina velutipes; D005656:Fungal Proteins; D007074:Immunoglobulin G; D007155:Immunologic Factors; D011993:Recombinant Fusion Proteins; D006632:Histamine; D000516:alpha-Amylases