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ARS Home Northeast Area Beltsville, Maryland (BARC) Beltsville Agricultural Research Center Molecular Plant Pathology Laboratory Docs







Cucumber Mosaic Virus-VLPs epitopeheadline bar ***Cucumber mosaic virus*****-VLPs epitope presentation systems for*Newcastle Disease Virus*(NDV) and*Avian Influenza virus*(AIV)** **** In order to utilize viruses in nanotechnology, they must be modified from their natural forms to impart new functions. This challenging process can be performed through several mechanisms including genetic modification and construction of virus particle chimeras which we developed as presentation systems for epitopes derived from other pathogens, such as animal viruses. Chimeric VLPs are constructed by genetic modification, inserting nucleotide sequences, resulting in a peptide insertion on the surface-exposed loop of the plant virus capsid protein.A potentially very important application is the development of novel or improved vaccines against animal viral diseases, especially in those cases where the use of the pathogens themselves for vaccination is unfeasible or unadvisable for technical or safety reasons.  /ARSUserFiles/80420560/cucumber1.jpg  Transmission electron microscopy of CMV VLPs, purified from*Nicotiana benthamiana*plants by standard method [6], fractionated by sucrose density gradient and concentrated by high-speed centrifugation. (A) CMV CP-WT; (B) CMV CP-F; (C) CMV CP-HN; (D) CMV CP-F/HN. Bars, 100 nm.   Reference Natilla, A., Hammond, RW., Nemchinov, L.G. 2006. Epitope presentation system based on cucumber mosaic virus coat protein expressed from a potato virus x-based vector. Archives of Virology. 15:1373-1386. Natilla A. and Nemchinov LG. Improvement of PVX/CMV CP expression tool for display of short foreign antigens. Protein Expr Purif. 59:117-21. 2008.    Among our accomplishments, VLPs derived from*Cucumber mosaic virus*(CMV) were engineered to carry antigenicdeterminant of *Newcastle disease virus*(NDV) proteins. The construct (vector and CMV coat protein engineered) was introduced into*Nicotiana benthamiana*plants. The recombinant CMV coat proteins (CPs) in plants formed VLPs carrying the selected antigenic determinant, which then had immune-reactivity with NDV. Immunization of chickens with these plant-derived molecules elicited the expected production of NDV antibodies. These findings show great potential for the development of vaccination against Newcastle disease, an economically important pathogen of poultry.   Additionally, we have been able to use the above CMV-VLPs platform, for the development of a potential vaccine candidate for*Avian Influenza virus.*Another economically important pathogen of poultry,*Avian Influenza**virus*is a highly contagious disease.  docs/23469_23695/image_4_smaller.jpg/ARSUserFiles/80420560/cucumber_pt2.jpg  Predicted secondary structures of the wild type CMV CP subunit and hybrid CMV CP with M2e epitope, respectively. Arrowsindicate ?H-?I loop (motif 5). Reference Nemchinov, L.G., Natilla, A. 2007. Transient expression of the ectodomain of matrix protein 2 (M2e) of Avian Influenza A Virus in plants. Protein Expression and Purification. 56:153-9. 2007. Available: doi:10.1016/j.pep.2007.05.015. ****


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