Journal: Cell Reports Medicine

Article Title: Persistence of MERS-CoV-spike-specific B cells and antibodies after late third immunization with the MVA-MERS-S vaccine

doi: 10.1016/j.xcrm.2022.100685

Figure Lengend Snippet: Study design Study participants (n = 10) received two vaccinations wit MVA-MERS-S (V1 and V2) 28 days apart and a late third vaccination (V3) at 12 ± 4 months after prime. Blood was frequently sampled for up to 2 years after V3 at indicated time points (see also Table S1 ). Figure was created with BioRender.com .

Article Snippet: MERS-CoV Spike S1+S2 Protein (ECD, aa 1-1297, His Tag), Biotinylated , Sino Biological , Cat#40069-V08B-B.

Techniques:

Journal: Cell Reports Medicine

Article Title: Persistence of MERS-CoV-spike-specific B cells and antibodies after late third immunization with the MVA-MERS-S vaccine

doi: 10.1016/j.xcrm.2022.100685

Figure Lengend Snippet: MERS-CoV-S-specific humoral and T cell responses induced by three vaccinations with MVA-MERS-S (A) Longitudinal dynamics of S-specific IgG antibodies. Shown are the optical density (OD) values measured at 450–620 nm by ELISA. Data are represented as individual data points (mean of technical duplicates) and median ± interquartile range (IQR). (B) Neutralization activity of serum antibodies as measured by 50% plaque-reduction neutralization test (PRNT50). Data are represented as individual data points and median ± IQR. (C) Spearman correlation between S-specific IgG antibodies and serum neutralization activity. (D) T cell responses as measured by IFNγ ELISpot after stimulation with five overlapping peptide (OLP) pools (M1–M5), spanning the entire MERS-CoV-S amino acid sequence. Shown are the median values of spot forming units (SFUs; mean of technical triplicates) across all vaccinees (n = 10) for each OLP pool. Number of samples, median, and p values for each time point and all three assays are shown in Table S2 .

Article Snippet: MERS-CoV Spike S1+S2 Protein (ECD, aa 1-1297, His Tag), Biotinylated , Sino Biological , Cat#40069-V08B-B.

Techniques: Enzyme-linked Immunosorbent Assay, Neutralization, Activity Assay, Plaque Reduction Neutralization Test, Enzyme-linked Immunospot, Sequencing

Journal: Cell Reports Medicine

Article Title: Persistence of MERS-CoV-spike-specific B cells and antibodies after late third immunization with the MVA-MERS-S vaccine

doi: 10.1016/j.xcrm.2022.100685

Figure Lengend Snippet: Isotype and subclass distribution within vaccine-induced MERS-CoV-S-specific antibodies S1- (A) and S2- (B) specific responses of IgM, IgG1–4, and IgA1–2 at different time points after vaccination, displayed as fold changes of median fluorescence intensities (MFIs; measured by bead-based ELISA) compared with baseline values at V1:day 0. Data are represented as individual data points (mean of technical duplicates) and median ± IQR. Number of samples and median fold changes are shown in Table S3 .

Article Snippet: MERS-CoV Spike S1+S2 Protein (ECD, aa 1-1297, His Tag), Biotinylated , Sino Biological , Cat#40069-V08B-B.

Techniques: Fluorescence, Enzyme-linked Immunosorbent Assay

Journal: Cell Reports Medicine

Article Title: Persistence of MERS-CoV-spike-specific B cells and antibodies after late third immunization with the MVA-MERS-S vaccine

doi: 10.1016/j.xcrm.2022.100685

Figure Lengend Snippet: Longitudinal dynamics of MERS-CoV-S-specific IgG1 and IgG3 antibodies Vaccine-induced S1/S2-specific IgG1 (A) and IgG3 (B) are displayed as MFI, measured by bead-based multiplex ELISA. Data are represented as individual data points (mean of technical duplicates) and median ± IQR. LLD, lower limit of detection. Number of samples, median, and p values for each time point are shown in Table S4 .

Article Snippet: MERS-CoV Spike S1+S2 Protein (ECD, aa 1-1297, His Tag), Biotinylated , Sino Biological , Cat#40069-V08B-B.

Techniques: Multiplex Assay, Enzyme-linked Immunosorbent Assay

Journal: Cell Reports Medicine

Article Title: Persistence of MERS-CoV-spike-specific B cells and antibodies after late third immunization with the MVA-MERS-S vaccine

doi: 10.1016/j.xcrm.2022.100685

Figure Lengend Snippet: Antigen-specific B cell responses induced by MVA-MERS-S vaccination (A) Representative IgG ELISpot images of antigen-specific and control wells for PBMCs taken before the first (V1:day 0) and after the third vaccination (V3:day 14). (B) Frequencies of vaccine-induced S1/S2-specific B cells displayed as SFUs/10 6 PBMCs as determined by IgG ELISpot. Data are represented as individual data points (mean of technical duplicates) and median ± IQR. The dotted line indicates the cutoff value (6.6 SFUs/10 6 PBMCs). (C) p values as determined by Wilcoxon signed rank test (between time points) and Mann Whitney U test (between S1 and S2 responses). Number of samples, median, and p values for each time point are shown in Table S5 .

Article Snippet: MERS-CoV Spike S1+S2 Protein (ECD, aa 1-1297, His Tag), Biotinylated , Sino Biological , Cat#40069-V08B-B.

Techniques: Enzyme-linked Immunospot, MANN-WHITNEY

Journal: Cell Reports Medicine

Article Title: Persistence of MERS-CoV-spike-specific B cells and antibodies after late third immunization with the MVA-MERS-S vaccine

doi: 10.1016/j.xcrm.2022.100685

Figure Lengend Snippet: Characterization of vaccine-induced memory B cells (A) Gating strategy for analysis of isotypes and MERS-CoV-S-specific cells within the memory B cell (MBC) population (representative contour plots belong to time point V3:day 14 from one study participant; gating strategy for identification of MBCs within whole PBMCs shown in Figure S2 ). (B) Longitudinal dynamics of antigen-specific MBCs induced by three vaccinations with MVA-MERS-S (V1, V2, and V3). Data are displayed as frequencies of S-specific cells within IgM + /IgG + /IgA + MBCs. Boxplots indicate median ± IQR. Number of samples, median, and p values compared with V1:day 0 are shown in Table S6 . (C) Resting, intermediate, atypical, and activated MBC phenotypes as identified by expression of CD21 and CD27 (top left panel). Representative plots are shown for one study participant at V3:day 0 and V3:day 14 and depict overlaid contour plots of total IgG + MBCs and S-specific IgG + MBCs (bottom left panel). Longitudinal distributions of phenotypes within the S-specific IgG + MBC compartment are shown as mean values of all study participants.

Article Snippet: MERS-CoV Spike S1+S2 Protein (ECD, aa 1-1297, His Tag), Biotinylated , Sino Biological , Cat#40069-V08B-B.

Techniques: Expressing

Journal: Cell Reports Medicine

Article Title: Persistence of MERS-CoV-spike-specific B cells and antibodies after late third immunization with the MVA-MERS-S vaccine

doi: 10.1016/j.xcrm.2022.100685

Figure Lengend Snippet: Correlation between vaccine-induced antibody and B cell responses Correlation analysis of MERS-CoV-S1- (A) and -S2- (B) specific IgG1, IgG3, and ASBC responses at different time points after vaccinations 1, 2, and 3 (V1, V2, and V3). S-specific MBC responses were included into correlograms of both the S1 and S2 antigen. Positive correlations are shown in blue and negative correlations in red, as indicated by the color bar. Sample numbers included into correlation analysis are provided in Table S7 .

Article Snippet: MERS-CoV Spike S1+S2 Protein (ECD, aa 1-1297, His Tag), Biotinylated , Sino Biological , Cat#40069-V08B-B.

Techniques:

Journal: Cell Reports Medicine

Article Title: Persistence of MERS-CoV-spike-specific B cells and antibodies after late third immunization with the MVA-MERS-S vaccine

doi: 10.1016/j.xcrm.2022.100685

Figure Lengend Snippet:

Article Snippet: MERS-CoV Spike S1+S2 Protein (ECD, aa 1-1297, His Tag), Biotinylated , Sino Biological , Cat#40069-V08B-B.

Techniques: Recombinant, Staining, Enzyme-linked Immunospot, Software

Journal: Cell Host & Microbe

Article Title: Rare, convergent antibodies targeting the stem helix broadly neutralize diverse betacoronaviruses

doi: 10.1016/j.chom.2022.10.010

Figure Lengend Snippet: Identification of broadly neutralizing antibodies against human betacoronaviruses The area under the curve (AUC) from titration of mAb binding to spike proteins from human betacoronaviruses SARS-CoV-2 Wuhan-Hu-1 (CoV-2), SARS-CoV (CoV-1), MERS-CoV, HCoV-HKU1, and HCoV-OC43, as well as alphacoronaviruses HCoV-NL63 and HCoV-229E, is shown on the panel of the left. Influenza H1 hemagglutinin (HA) was included as a control antigen and L9 IgG1 (malaria specific; Wang et al., 2020 ) was included as a negative control mAb for binding experiments. AUC values for each antigen are shown after subtraction with values for the negative control antigen CD4. The antibody titers at 50% neutralization (NT 50 ) against SARS-CoV-2 Wuhan-Hu-1, SARS-CoV, MERS-CoV, HCoV-NL63 envelope-pseudotyped virus, as well as authentic HCoV-OC43, are shown on the right. Neutralizing mAbs are ranked by their breadth of neutralization and the geometric mean of their NT 50 values. Cells highlighted in blue denote mAbs that did not show neutralizing potency at the highest concentration tested (100 μg/mL). Negative control mAbs for neutralization are DEN3 (dengue-specific; Rogers et al., 2020 ) for SARS-CoV-2, SARS-CoV, MERS-CoV and HCoV-NL63, and CV503 (SARS-CoV-2 RBD-specific; Cho et al., 2021 ) for HCoV-OC43. NT 50 values were calculated using the dose-response-inhibition model with 5-parameter Hill slope equation in GraphPad Prism version 9.3.1.

Article Snippet: 229E-CoV spike , Sino Biological , Cat. # 40605-V08B-B.

Techniques: Titration, Binding Assay, Negative Control, Neutralization, Concentration Assay, Inhibition

Journal: Cell Host & Microbe

Article Title: Rare, convergent antibodies targeting the stem helix broadly neutralize diverse betacoronaviruses

doi: 10.1016/j.chom.2022.10.010

Figure Lengend Snippet:

Article Snippet: 229E-CoV spike , Sino Biological , Cat. # 40605-V08B-B.

Techniques: Recombinant, Transfection, Lysis, Cell Culture, Luciferase, Expressing, Irradiation, Amplification, Plasmid Preparation, Software, Fluorescence, Microscopy, Imaging