Journal: Frontiers in Immunology

Article Title: Distinct Immune Response at 1 Year Post-COVID-19 According to Disease Severity

doi: 10.3389/fimmu.2022.830433

Figure Lengend Snippet: Longitudinal analysis of anti-SARS-CoV-2 antibody in COVID-19 patients over time according to the severity of illness. (A) Schematic illustration (created with BioRender.com ) of experimental design for analyzing humoral and cellular immune response to SARS-CoV-2 according to the disease severity, including Figures 1 – 4 . (B) Binding activities of each Ig isotype to SARS-CoV-2 proteins. Diluted plasma samples (1:100) in PBS were incubated with SARS-CoV-2 spike protein, receptor-binding domain (RBD), nucleocapsid (NC), and M protein. Corresponding isotyping Abs determined binding activities of IgM, IgA, and IgG to SARS-CoV-2 antigens. ASx, asymptomatic (8-month: n = 7, 12-month: n = 6), Mild (8-month: n = 2, 12-month: n = 9), Severe (8-month: n = 7, 12-month: n = 8). Statistical analyses were performed using the Kruskal–Wallis rank-sum test with Dunn’s post hoc test in GraphPad Prism ( n.s .: P > 0.05, * P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: Other SARS-CoV-2 antigens, namely, spike (Sino Biological, Wayne, PA, USA), nucleocapsid (NC; Prosci, Fort Collins, CO, USA), and membrane (M) protein (MRC PPU, Dundee, UK), were purchased.

Techniques: Binding Assay, Clinical Proteomics, Incubation

Journal: Viruses

Article Title: NSs Filament Formation Is Important but Not Sufficient for RVFV Virulence In Vivo

doi: 10.3390/v11090834

Figure Lengend Snippet: Rescue of RVFV BJ01 and a mutant lacking the NSs gene. ( A ) Schematic of the T7 RNA polymerase-dependent reverse genetics system for construction of recombinant viruses Adapted from the Box 2 figure of Orthobunyaviruses: recent genetic and structural insights . ( B ) Cytopathic effect (CPE) of rescued virus in BHK-21 cells. BHK-21 cells were mock treated or infected with rWT at an MOI of 5, images were captured at 48 h p.i. Bars, 20 μm. ( C ) Construction of the plasmid encoding the S segment in which the NSs gene was replaced by eGFP. ( D ) Fluorescence microscopy analysis to verify the successful rescue of r△NSs-eGFP. BHK-21 cells were infected with r△NSs-eGFP (MOI = 5) and the fluorescence images were acquired at 72 h p.i. Bars, 20 μm. ( E ) Western blot analysis of infected cells with anti-N, anti-NSs and anti-Tubulin antibodies. Vero cells were infected with WT, rWT and r△NSs-eGFP at an MOI of 1 and cell lysates were harvested at 24 h p.i. ( F ) Plaque assay of RVFV WT and rescued viruses (rWT and r△NSs-eGFP) on Vero cells. After 7 days of incubation, plaques were stained with crystal violet.

Article Snippet: Proteins were incubated with primary antibodies against nucleocapsid protein (NP), NSs and Tubulin (Abclonal, Wuhan, China), then secondary horseradish peroxidase-conjugated goat anti-rabbit IgG.

Techniques: Mutagenesis, Recombinant, Virus, Infection, Plasmid Preparation, Fluorescence, Microscopy, Western Blot, Plaque Assay, Incubation, Staining