Journal: PLOS ONE

Article Title: Discovery of anti-SARS-CoV-2 S2 protein antibody CV804 with broad-spectrum reactivity with various beta coronaviruses and analysis of its pharmacological properties in vitro and in vivo

doi: 10.1371/journal.pone.0300297

Figure Lengend Snippet: (A) Five humanized CV804 antibodies were analyzed by flow cytometric assay of titrated candidates against SARS-CoV-2 spike protein expressing cells. Four parameter logistic curve fitted with EC 50 in parentheses. Normalized by secondary alone and saturation signal. Representative data of two independent experiments are shown. (B) Evaluation of hCV804-40 antibody, REGN10987, and LY-CoV1404 antibody binding to cells infected with SARS-CoV-2 variants. "+++", "++", "+" indicate positive binding; "-" indicates negative binding. (C) Evaluation of antibody-dependent cell-mediated cytotoxicity ADCC activity against spike-expressing cells for mouse derived antibodies. (D) In vitro cell infection inhibition experiments were conducted. The viral strains used were hCoV-19/Japan/TY/WK-521/2020 (top) and SARS-CoV-2/Japan/TY-501/2020 (bottom), with VeroE6/TMPRSS2 cells being used as the target cells. The hCV804-35 antibody, REGN10987, and LY-CoV 1404 were measured in this experiment. The antibody concentration that inhibited cell death by 50% was defined as the IC 50 .

Article Snippet: ADCC activity was measured by mouse FcγRIV ADCC Reporter Bioassay kit (Promega) according to manufacturer’s instruction.

Techniques: Flow Cytometry, Expressing, Binding Assay, Infection, Activity Assay, Derivative Assay, In Vitro, Inhibition, Concentration Assay

Journal: NPJ Vaccines

Article Title: Preclinical evaluation of a universal inactivated influenza B vaccine based on the mosaic hemagglutinin-approach

doi: 10.1038/s41541-024-01014-8

Figure Lengend Snippet: Whole inactivated virus (WIV) with beta-propiolactone (BPL) and inactivated split vaccine combining BPL inactivation and Triton X-100 splitting were compared for two different vaccinations with WT viruses or mHA viruses. Vaccines were tested without adjuvant or with the addition of CpG 1018 (30 µg) as explained in Fig. . A Heatmap of IgG binding serum antibodies against a panel of recombinant influenza B HA proteins. B Heatmap of hemagglutination inhibition (HI) titer against a panel of influenza B viruses. C Heatmap of ADCC activity against three influenza B viruses. To perform the ADCC reporter assay, MDCK cells were infected with each virus at an MOI of 5 (single-cycle replication). One day after infection, mouse sera were added to the cells and incubated for 30 min and genetically modified Jurkat cells expressing the mouse FcγRIV with a luciferase reporter gene under transcriptional control of the nuclear factor-activated T (NFAT) cell promoter were then added and incubated for 6 h. Later luciferase activity was measured. The geometric mean AUC of fold induction of each group (pooled sera) was measured in technical duplicate.

Article Snippet: Genetically modified Jurkat cells expressing the mouse FcγRIV with a luciferase reporter gene under transcriptional control of the nuclear factor-activated T (NFAT) cell promoter were then added to the plate at 7.5× 10 4 cells in 25 μL/well (Promega) and incubated for 6 hours at 37 °C.

Techniques: Virus, Vaccines, Adjuvant, Binding Assay, Recombinant, HI Assay, Activity Assay, Reporter Assay, Infection, Incubation, Genetically Modified, Expressing, Luciferase, Control