Abstract
A strain-specific vaccine represents the best possible response to the threat of an influenza pandemic. Rapid delivery of such a vaccine to the world's population before the peak of the first infection wave seems to be an unattainable goal with the current influenza vaccine manufacturing capacity. Plant-based transient expression is one of the few production systems that can meet the anticipated surge requirement. To assess the capability of plant agroinfiltration to produce an influenza vaccine, we expressed haemagglutinin (HA) from strains A/Indonesia/5/05 (H5N1) and A/New Caledonia/20/99 (H1N1) by agroinfiltration of Nicotiana benthamiana plants. Size distribution analysis of protein content in infiltrated leaves revealed that HA was predominantly assembled into high-molecular-weight structures. H5-containing structures were purified and examination by transmission electron microscopy confirmed virus-like particle (VLP) assembly. High-performance thin layer chromatography analysis of VLP lipid composition highlighted polar and neutral lipid contents comparable with those of purified plasma membranes from tobacco plants. Electron microscopy of VLP-producing cells in N. benthamiana leaves confirmed that VLPs accumulated in apoplastic indentations of the plasma membrane. Finally, immunization of mice with two doses of as little as 0.1 microg of purified influenza H5-VLPs triggered a strong immune response against the homologous virus, whereas two doses of 0.5 microg of H5-VLPs conferred complete protection against a lethal challenge with the heterologous A/Vietnam/1194/04 (H5N1) strain. These results show, for the first time, that plants are capable of producing enveloped influenza VLPs budding from the plasma membrane; such VLPs represent very promising candidates for vaccination against influenza pandemic strains.
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Claims 8
Claim · model-written summary
Haemagglutinin in agroinfiltrated Nicotiana benthamiana leaves was predominantly assembled into high-molecular-weight structures.
- Taxon
- Nicotiana benthamiana
Nicotiana benthamianae0 - Organ / stage
- infiltrated leavese1
- Gene
- haemagglutinin (HA)e0
- Outcome
- predominantly assembled into high-molecular-weight structurese1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“we expressed haemagglutinin (HA) from strains A/Indonesia/5/05 (H5N1) and A/New Caledonia/20/99 (H1N1) by agroinfiltration of Nicotiana benthamiana plants”
e1“Size distribution analysis of protein content in infiltrated leaves revealed that HA was predominantly assembled into high-molecular-weight structures”
Claim · model-written summary
Transmission electron microscopy confirmed virus-like particle assembly in purified H5-containing structures.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Outcome
- virus-like particle (VLP) assemblye2
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“we expressed haemagglutinin (HA) from strains A/Indonesia/5/05 (H5N1) and A/New Caledonia/20/99 (H1N1) by agroinfiltration of Nicotiana benthamiana plants”
e2“H5-containing structures were purified and examination by transmission electron microscopy confirmed virus-like particle (VLP) assembly”
Claim · model-written summary
The lipid composition of the VLPs had polar and neutral lipid contents comparable to purified tobacco plasma membranes.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Outcome
- polar and neutral lipid contentse3
- Comparator
- purified plasma membranes from tobacco plantse3
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“we expressed haemagglutinin (HA) from strains A/Indonesia/5/05 (H5N1) and A/New Caledonia/20/99 (H1N1) by agroinfiltration of Nicotiana benthamiana plants”
e3“High-performance thin layer chromatography analysis of VLP lipid composition highlighted polar and neutral lipid contents comparable with those of purified plasma membranes from tobacco plants”
Claim · model-written summary
VLPs accumulated in apoplastic indentations of the plasma membrane of Nicotiana benthamiana leaf cells.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Organ / stage
- VLP-producing cellse4
- Outcome
- VLPs accumulated in apoplastic indentations of the plasma membranee4
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“we expressed haemagglutinin (HA) from strains A/Indonesia/5/05 (H5N1) and A/New Caledonia/20/99 (H1N1) by agroinfiltration of Nicotiana benthamiana plants”
e4“Electron microscopy of VLP-producing cells in N. benthamiana leaves confirmed that VLPs accumulated in apoplastic indentations of the plasma membrane”
Claim · model-written summary
Two doses of as little as 0.1 microg of purified H5-VLPs triggered a strong immune response in mice against the homologous virus.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Treatment
- two doses of as little as 0.1 microg of purified influenza H5-VLPse5
- Outcome
- strong immune response against the homologous viruse5
- Direction
- triggerede5
- Qualifier
- micee5
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“we expressed haemagglutinin (HA) from strains A/Indonesia/5/05 (H5N1) and A/New Caledonia/20/99 (H1N1) by agroinfiltration of Nicotiana benthamiana plants”
e5“immunization of mice with two doses of as little as 0.1 microg of purified influenza H5-VLPs triggered a strong immune response against the homologous virus”
Claim · model-written summary
Two doses of 0.5 microg of H5-VLPs conferred complete protection in mice against a lethal challenge with the heterologous A/Vietnam/1194/04 (H5N1) strain.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Treatment
- two doses of 0.5 microg of H5-VLPse6
- Interactor
- A/Vietnam/1194/04 (H5N1)
_virion%2C_a_type_of_bird_flu_virus_which_is_a_subtype_of_avian_influenza_A.jpg?width=60)
H5N1 subtypee6pathogen - Outcome
- complete protection against a lethal challengee6
- Direction
- conferrede6
- Qualifier
- heterologouse6
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
_virion%2C_a_type_of_bird_flu_virus_which_is_a_subtype_of_avian_influenza_A.jpg?width=500)
Verbatim
e0“we expressed haemagglutinin (HA) from strains A/Indonesia/5/05 (H5N1) and A/New Caledonia/20/99 (H1N1) by agroinfiltration of Nicotiana benthamiana plants”
e6“two doses of 0.5 microg of H5-VLPs conferred complete protection against a lethal challenge with the heterologous A/Vietnam/1194/04 (H5N1) strain”
Claim · model-written summary
Plants can produce enveloped influenza VLPs that bud from the plasma membrane.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Outcome
- producing enveloped influenza VLPs budding from the plasma membranee7
- Qualifier
- for the first timee7
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“we expressed haemagglutinin (HA) from strains A/Indonesia/5/05 (H5N1) and A/New Caledonia/20/99 (H1N1) by agroinfiltration of Nicotiana benthamiana plants”
e7“These results show, for the first time, that plants are capable of producing enveloped influenza VLPs budding from the plasma membrane”
Claim · model-written summary
The authors consider plant-produced H5 VLPs very promising candidates for vaccination against influenza pandemic strains.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Outcome
- candidates for vaccination against influenza pandemic strainse1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“we expressed haemagglutinin (HA) from strains A/Indonesia/5/05 (H5N1) and A/New Caledonia/20/99 (H1N1) by agroinfiltration of Nicotiana benthamiana plants”
e1“such VLPs represent very promising candidates for vaccination against influenza pandemic strains”