Characterization of an immunomodulatory Der p 2-FIP-fve fusion protein produced in transformed rice suspension cell culture |
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Authors: | Chin-Fen Su I-Chun Kuo Peng-Wen Chen Chiung-Hui Huang See Voon Seow Kaw Yan Chua Su-May Yu |
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Institution: | (1) Institute of Biotechnology, National Cheng Kung University, No.1, University Road, Tainan 701, Taiwan, ROC;(2) Department of Paediatrics, Yong Loo Lin School of Medicine, National University of Singapore, 5 Lower Kent Ridge Road, Singapore, Singapore, 119074;(3) Department and Institute of Bioagricultural Sciences, National Chiayi University, Chiayi 600, Taiwan, ROC;(4) Institute of Molecular Biology, Academia Sinica, Nankang, Taipei 115, Taiwan, ROC; |
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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. |
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