Journal: mBio

Article Title: A decavalent composite mRNA vaccine against both influenza and COVID-19

doi: 10.1128/mbio.00668-24

Figure Lengend Snippet: Design and characterization of 10-valent mRNA vaccine (FLUCOV-10). ( A ). Schematic illustration of the FLUCOV-10 formulation, a 10-valent combination mRNA vaccine targeting both influenza and COVID-19. It includes mRNAs encoding the full-length HA proteins from influenza A virus subtypes A/H1, A/H3, A/H5, and A/H7, and from influenza B virus lineages B/Yamagata and B/Victoria. Additionally, it encodes full-length spike proteins from SARS-CoV-2 variants Wuhan-Hu-1, BQ.1.1, BA.2.75.2, and XBB.1.5. Each mRNA component is individually encapsulated in lipid nanoparticles (LNPs) prior to being combined into the final FLUCOV-10 formulation. (B and C) Phylogenic trees were created for influenza HAs ( B ) and SARS-CoV-2 spikes ( C ) by using Nextstrain. The vaccine HAs or spike are indicated with red triangles, and the challenge viruses are indicated with an “X.” (D) The expression of FLUCOV-10-mRNA-encoded HA proteins in 293T cells was determined by western blotting. Lane 1, 293T cells with mock transfection; lane 2, 293T cells with indicated mRNA transfection. (E) The expression of FLUCOV-10-mRNA-encoded spike proteins. Lane 1, 293T cells with mock transfection; lane 2–5, 293T cells with Wuhan-Hu-1, BQ.1.1, BA.2.75.2, and XBB.1.5 mRNA transfection, respectively. β-Actin was used as western blotting loading control. (F) Particle size distribution of individual LNP formulations was measured in triplicate using dynamic light scattering (DLS) on the NS-90Z Nanoparticle Size and Zeta Potential Analyzer.

Article Snippet: The HA and spike proteins in cell lysates were then detected by western blotting using a mouse monoclonal antibody against SARS-CoV-2 spike proteins (GTX632604, GeneTex), a rabbit polyclonal antibody against influenza A/H1 HA (11055-T62, Sino Biological), a mouse monoclonal antibody against influenza A/H3 HA (11056-MM03, Sino Biological), a rabbit polyclonal antibody against influenza A/H5 HA (11062-T62, Sino Biological), a rabbit polyclonal antibody against influenza A/H7 HA (40103-T62, Sino Biological), a rabbit polyclonal antibody against influenza B/Yamgata lineage HA (11053-T62, Sino Biological), and a mouse monoclonal antibody against Influenza B/Victoria lineage HA (11053-MM06, Sino Biological). β-Actin was detected using anti-β-actin antibody.

Techniques: Formulation, Virus, Expressing, Western Blot, Transfection, Control, Zeta Potential Analyzer

Journal: Virology

Article Title: Identification of key hemagglutinin residues responsible for cleavage, acid stability, and virulence of fifth-wave highly pathogenic avian influenza A(H7N9) viruses

doi: 10.1016/j.virol.2019.07.012

Figure Lengend Snippet: The TW/17 HA trimer was modeled with PyMOL version 2.2.3 from A/Guangdong/17SF003/2016 (PDB ID: 6D7U) with the highlighted residues HA1-(38, 112, 164, 217; H7 numbering), and HA2-64 in one monomer (left) and a close-up image of the residue HA2-K64 forming potential hydrogen bonds with HA2-N79 and the glycans of HA2-N82 from a neighboring monomer (right).

Article Snippet: At 3h post fusion induction, syncytia were visualized by immunofluorescence microscopy after staining the cells with rabbit anti -H7 HA monoclonal antibody (Sino Biological, Cat#11082-R002, PA) and Alexa Fluor™ 488-labeled anti-rabbit IgG secondary antibody (Thermo Fisher Scientific), and diamidino-2-phenylindole (DAPI) for nuclei.

Techniques: