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

Abstract

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.Electronic supplementary materialThe online version of this article (doi:10.1186/s12896-015-0164-6) contains supplementary material, which is available to authorized users.

Supplemented vaccination with tandem repeat M2e virus-like particles enhances protection against homologous and heterologous HPAI H5 viruses in chickens

“…Previous studies of M2e-based vaccines demonstrated substantially different results depending on the experimental animal model used. The use of M2e-based vaccines alone in mouse models confers protection against HPAI and low pathogenic avian influenza (LPAI) virus challenge, making them candidate universal influenza A vaccines [6, 16, 28, 29]. However, challenge studies in chickens have demonstrated protection against LPAI but not HPAI, although antibodies elicited by M2e-based vaccines were induced in immunized chickens [7, 30, 31].…”

Synthetic Botany

“…This allows relatively fast characterization of libraries of genetic components in a common genetic background (Sparkes et al 2006;Engler et al 2014;Brückner et al 2015). Transient leaf agroinfiltration in Nicotiana has been adopted as an efficient method for the optimization of metabolic pathways, such as artemisinin (van Herpen et al 2010) and triterpene biosynthesis pathways (reviewed in Thimmappa et al 2014), and the bioproduction of vaccines (D'Aoust et al 2008, Mardanova et al 2015. Large investments have been made in both the private and the public sectors to scale up production for vaccines against viruses like Ebola (ZMapp, Mapp Biopharmaceuticals) and influenza (Medicago).…”

Current Approaches to Universal Vaccine Against Influenza Virus

“…с соавт. продемонстрировала иммуногенность и протективность химерного белка, состоящего из 4 аминокислотных по-следовательностей М2е (2 последовательнос-ти М2е консенсусные между человеческими штамма вируса гриппа А и 2 последовательнос-ти М2е от штамма A/Chicken/Kurgan/05/2005), соединенных с аминокислотной последова-тельностью белка флаггелина от Salmonella typhimurium [60]. Данная химерная конструк-ция была получена при помощи экспрессии плазмидного вектора в растении Nicotiana benthamiana.…”

“…Данная химерная конструк-ция была получена при помощи экспрессии плазмидного вектора в растении Nicotiana benthamiana. Трехкратная иммунизация мы-шей 10 мкг полученного препарата индуцирует выработку М2е-специфичных антител и обес-печивает 75%-ную защиту против 5ЛД 50 [60]. Имеются данные о том, что кандидатная уни-версальная вакцина на основе химерных но-ровирусных частиц, модифицированных кон-сенсусным птичьим М2е (М2е-РР), способна индуцировать выработку значимых титров IgG у кур при подкожном введении в сочетании с масляным адъювантом [16].…”