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Vaccine v.36 no.16, 2018년, pp.2147 - 2154   SCI SCIE SCOPUS
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Morphological characterization of a plant-made virus-like particle vaccine bearing influenza virus hemagglutinins by electron microscopy

Lindsay, Brianne J. (Department of Anatomy & Cell Biology, Faculty of Medicine, McGill University, Groupe de Recherche Axe sur la Structure des Proteines (GRASP), Groupe d'Etude de Proteines Membranaires (GEPROM), 3640 University Street, Montréal, QC H3A 2B2, Canada ) ; Bonar, Michal M. (Department of Anatomy & Cell Biology, Faculty of Medicine, McGill University, Groupe de Recherche Axe sur la Structure des Proteines (GRASP), Groupe d'Etude de Proteines Membranaires (GEPROM), 3640 University Street, Montréal, QC H3A 2B2, Canada ) ; Costas-Cancelas, Ian N. (Department of Anatomy & Cell Biology, Faculty of Medicine, McGill University, Groupe de Recherche Axe sur la Structure des Proteines (GRASP), Groupe d'Etude de Proteines Membranaires (GEPROM), 3640 University Street, Montréal, QC H3A 2B2, Canada ) ; Hunt, Kristin (Department of Microbiology and Immunology, and Physiology, Goodman Cancer Research Center, McGill University, Montréal, QC, Canada ) ; Makarkov, Alexander I. (Research Institute of McGill University Health Centre-Suite EM3-3248, 1001 Decarie Street ) ; Chierzi, Sabrina ; Krawczyk, Connie M. ; Landry, Nathalie ; Ward, Brian J. ; Rouiller, Isabelle ;
  • 초록  

    Abstract Plant-made virus-like particle (VLP) vaccines that display wild-type influenza hemagglutinin (HA) are rapidly advancing through clinical trials. Produced by transient transfection of Nicotiana benthamiana , these novel vaccines are unusually immunogenic, eliciting both humoral and cellular responses. Here, we directly visualized VLPs bearing either HA trimers derived from strains A/California/7/2009 or A/Indonesia/5/05 using cryo-electron microscopy and determined the 3D organization of the VLPs using cryo-electron tomography. More than 99.9% of the HA trimers in the vaccine preparations were found on discoid and ovoid-shaped particles. The discoid-shaped VLPs presented HA trimers on their outer diameter. The ovoid-shaped VLPs contained HA trimers evenly distributed at their surface. The VLPs were stable for 12 months at 4 °C. Early interactions of the VLPs with mouse dendritic and human monocytoid (U-937) cells were visualized by electron microscopy after resin-embedding and sectioning. The VLP particles were observed bound to plasma membranes as well as inside vesicles. Mouse dendritic cells exposed to VLPs displayed classic morphological changes associated with activation including the extensive formation of dendrites. Our findings demonstrate that plant-made VLPs bearing influenza HA trimers are morphologically stable over time and raise the possibility that these VLPs may interact with and activate antigen-presenting cells in a manner similar to the intact virus. Highlights Analysis of a plant made VLP vaccine candidate using cryo-EM and tomography. 99.9% of the HA trimers are on discoid and ovoid-shaped particles. VLPs remain structurally stable for over 12 months while kept at 4 °C. VLPs bind to plasma membrane and are internalized similarly to intact virus. Mouse dendritic cells undergo morphological changes indicative activation.


  • 주제어

    Plant-made virus-like particle .   Cryo-electron microscopy .   Phase plate tomography .   Hemagglutinin .   Antigen presenting cells .   Cellular activation.  

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