High-Level Systemic Expression of Conserved Influenza Epitope in Plants on the Surface of Rod-Shaped Chimeric Particles
Recombinant viruses based on the cDNA copy of the tobacco mosaic virus (TMV) genome carrying different versions of the conserved M2e epitope from influenza virus A cloned into the coat protein (CP) gene were obtained and partially characterized by our group previously; cysteines in the human consens...
Main Authors: | , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
MDPI AG
2014-04-01
|
Series: | Viruses |
Subjects: | |
Online Access: | http://www.mdpi.com/1999-4915/6/4/1789 |
id |
doaj-13efb4e768954ca38e86e20c5635da77 |
---|---|
record_format |
Article |
spelling |
doaj-13efb4e768954ca38e86e20c5635da772020-11-25T00:00:24ZengMDPI AGViruses1999-49152014-04-01641789180010.3390/v6041789v6041789High-Level Systemic Expression of Conserved Influenza Epitope in Plants on the Surface of Rod-Shaped Chimeric ParticlesNatalia V. Petukhova0Tatiana V. Gasanova1Peter A. Ivanov2Joseph G. Atabekov3Department of Virology, Lomonosov Moscow State University, Moscow 119991, RussiaDepartment of Virology, Lomonosov Moscow State University, Moscow 119991, RussiaDepartment of Virology, Lomonosov Moscow State University, Moscow 119991, RussiaDepartment of Virology, Lomonosov Moscow State University, Moscow 119991, RussiaRecombinant viruses based on the cDNA copy of the tobacco mosaic virus (TMV) genome carrying different versions of the conserved M2e epitope from influenza virus A cloned into the coat protein (CP) gene were obtained and partially characterized by our group previously; cysteines in the human consensus M2e sequence were changed to serine residues. This work intends to show some biological properties of these viruses following plant infections. Agroinfiltration experiments on Nicotiana benthamiana confirmed the efficient systemic expression of M2e peptides, and two point amino acid substitutions in recombinant CPs significantly influenced the symptoms and development of viral infections. Joint expression of RNA interference suppressor protein p19 from tomato bushy stunt virus (TBSV) did not affect the accumulation of CP-M2e-ser recombinant protein in non-inoculated leaves. RT-PCR analysis of RNA isolated from either infected leaves or purified TMV-M2e particles proved the genetic stability of TMV‑based viral vectors. Immunoelectron microscopy of crude plant extracts demonstrated that foreign epitopes are located on the surface of chimeric virions. The rod‑shaped geometry of plant-produced M2e epitopes is different from the icosahedral or helical filamentous arrangement of M2e antigens on the carrier virus-like particles (VLP) described earlier. Thereby, we created a simple and efficient system that employs agrobacteria and plant viral vectors in order to produce a candidate broad-spectrum flu vaccine.http://www.mdpi.com/1999-4915/6/4/1789influenza virusM2eplanttobacco mosaic virussystemic expressioncoat protein fusion |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Natalia V. Petukhova Tatiana V. Gasanova Peter A. Ivanov Joseph G. Atabekov |
spellingShingle |
Natalia V. Petukhova Tatiana V. Gasanova Peter A. Ivanov Joseph G. Atabekov High-Level Systemic Expression of Conserved Influenza Epitope in Plants on the Surface of Rod-Shaped Chimeric Particles Viruses influenza virus M2e plant tobacco mosaic virus systemic expression coat protein fusion |
author_facet |
Natalia V. Petukhova Tatiana V. Gasanova Peter A. Ivanov Joseph G. Atabekov |
author_sort |
Natalia V. Petukhova |
title |
High-Level Systemic Expression of Conserved Influenza Epitope in Plants on the Surface of Rod-Shaped Chimeric Particles |
title_short |
High-Level Systemic Expression of Conserved Influenza Epitope in Plants on the Surface of Rod-Shaped Chimeric Particles |
title_full |
High-Level Systemic Expression of Conserved Influenza Epitope in Plants on the Surface of Rod-Shaped Chimeric Particles |
title_fullStr |
High-Level Systemic Expression of Conserved Influenza Epitope in Plants on the Surface of Rod-Shaped Chimeric Particles |
title_full_unstemmed |
High-Level Systemic Expression of Conserved Influenza Epitope in Plants on the Surface of Rod-Shaped Chimeric Particles |
title_sort |
high-level systemic expression of conserved influenza epitope in plants on the surface of rod-shaped chimeric particles |
publisher |
MDPI AG |
series |
Viruses |
issn |
1999-4915 |
publishDate |
2014-04-01 |
description |
Recombinant viruses based on the cDNA copy of the tobacco mosaic virus (TMV) genome carrying different versions of the conserved M2e epitope from influenza virus A cloned into the coat protein (CP) gene were obtained and partially characterized by our group previously; cysteines in the human consensus M2e sequence were changed to serine residues. This work intends to show some biological properties of these viruses following plant infections. Agroinfiltration experiments on Nicotiana benthamiana confirmed the efficient systemic expression of M2e peptides, and two point amino acid substitutions in recombinant CPs significantly influenced the symptoms and development of viral infections. Joint expression of RNA interference suppressor protein p19 from tomato bushy stunt virus (TBSV) did not affect the accumulation of CP-M2e-ser recombinant protein in non-inoculated leaves. RT-PCR analysis of RNA isolated from either infected leaves or purified TMV-M2e particles proved the genetic stability of TMV‑based viral vectors. Immunoelectron microscopy of crude plant extracts demonstrated that foreign epitopes are located on the surface of chimeric virions. The rod‑shaped geometry of plant-produced M2e epitopes is different from the icosahedral or helical filamentous arrangement of M2e antigens on the carrier virus-like particles (VLP) described earlier. Thereby, we created a simple and efficient system that employs agrobacteria and plant viral vectors in order to produce a candidate broad-spectrum flu vaccine. |
topic |
influenza virus M2e plant tobacco mosaic virus systemic expression coat protein fusion |
url |
http://www.mdpi.com/1999-4915/6/4/1789 |
work_keys_str_mv |
AT nataliavpetukhova highlevelsystemicexpressionofconservedinfluenzaepitopeinplantsonthesurfaceofrodshapedchimericparticles AT tatianavgasanova highlevelsystemicexpressionofconservedinfluenzaepitopeinplantsonthesurfaceofrodshapedchimericparticles AT peteraivanov highlevelsystemicexpressionofconservedinfluenzaepitopeinplantsonthesurfaceofrodshapedchimericparticles AT josephgatabekov highlevelsystemicexpressionofconservedinfluenzaepitopeinplantsonthesurfaceofrodshapedchimericparticles |
_version_ |
1725445276235202560 |