Rapid high-yield expression of a candidate influenza vaccine based on the ectodomain of M2 protein linked to flagellin in plants using viral vectors

BMC Biotechnology

Research Article

Nikolai V. Ravin¹  Roman Y. Kotlyarov¹  Victor V. Kuprianov¹  Eugenia S. Mardanova¹  George P. Lomonosoff²  Liudmila A. Stepanova³  Liudmila M. Tsybalova³ 

[1] Centre ‘Bioengineering’, Russian Academy of Sciences, Prosp. 60-letya Oktyabrya, bld 7-1, 117312, Moscow, Russia;Department of Biological Chemistry, John Innes Centre, Norwich Research Park, NR4 7UH, Norwich, UK;Research Institute of Influenza, Russian Federation Ministry of Health, 15/17 Prof. Popova str., 197376, St. Petersburg, Russia;

关键词: Viral vector;    Potato virus X;    M2 protein;    Flagellin;    Influenza;    Vaccine;   

DOI  :  10.1186/s12896-015-0164-6

 received in 2015-02-20, accepted in 2015-05-15,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundThe extracellular domain of matrix protein 2 (M2e) of influenza A virus is a promising target for the development of a universal vaccine against influenza because M2e sequences are highly conserved among human influenza A strains. However, native M2e is poorly immunogenic, but its immunogenicity can be increased by delivery in combination with adjuvants or carrier particles. It was previously shown that fusion of M2e to bacterial flagellin, the ligand for Toll-like receptor (TLR) 5 and powerful mucosal adjuvant, significantly increases the immunogenicity and protective capacity of M2e.ResultsIn this study, we report for the first time the transient expression in plants of a recombinant protein Flg-4M comprising flagellin of Salmonella typhimurium fused to four tandem copies of the M2e peptide. The chimeric construct was expressed in Nicotiana benthamiana plants using either the self-replicating potato virus X (PVX) based vector, pA7248AMV-GFP, or the cowpea mosaic virus (CPMV)-derived expression vector, pEAQ-HT. The highest expression level up to 30 % of total soluble protein (about 1 mg/g of fresh leaf tissue) was achieved with the PVX-based expression system. Intranasal immunization of mice with purified Flg-4M protein induced high levels of M2e-specific serum antibodies and provided protection against lethal challenge with influenza virus.ConclusionsThis study confirms the usefulness of flagellin as a carrier of M2e and its relevance for the production of M2e-based candidate influenza vaccines in plants.

【 授权许可】

Unknown   
© Mardanova et al.; licensee BioMed Central. 2015. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.