Development of Tobacco necrosis virus A as a vector for efficient and stable expression of FMDV VP1 peptides
2010
Zhang, Yongliang | Li, Jiang | Pu, Heng | Jin, Jin | Zhang, Xiaofeng | Chen, Mingkuan | Wang, Bin | Han, Chenggui | Yu, Jialin | Li, Dawei
Plant virus-based expression systems provide attractive alternatives for production of animal virus-originated antigenic peptides. In the present study, an infectious cDNA clone of Tobacco necrosis virus A Chinese isolate (TNV-AC) was used for expression of different peptides derived from Foot and mouth disease virus (FMDV) serotype O VP1 fused downstream of the coat protein (CP) open reading frame (ORF). Chenopodium amaranticolor inoculated with in vitro transcripts of the chimaeras developed symptoms similar to those caused by wild-type TNV-AC. Western blot and RT-PCR detection of the infected leaves demonstrated that the chimaeras were infective, and a large number of self-assembled virions could be purified and observed under electron microscopy. Immunogold labelling revealed that highly expressed FMDV VP1 peptides could be displayed on the surfaces of virus particles. Additional immunoblotting and DNA sequence analyses showed that most of the chimaeras contained unmodified foreign peptides even after six successive passages in C. amaranticolor and three passages in Nicotiana benthamiana. Our results also suggest that the amino acid sequence and peptide length have a substantial influence on viral morphogenesis and systemic infections. Finally, animal experiments showed that purified chimaeric virus particles (CVPs) could induce a strong immune response against FMDV structural protein VP1 via an intramuscular route. And when inoculated nasally, CVPs could induce systemic and mucosal immune responses in mice.
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