mers cov nucleocapsid protein antibody rabbit pab  (Sino Biological)


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    Name:
    MERS CoV Nucleocapsid Protein Antibody Rabbit PAb
    Description:
    Produced in rabbits immunized with purified a synthetic peptide corresponding to the center region of MERS CoV NCoV Novel coronavirus Nucleocapsid Protein NP protein and purified by antigen affinity chromatography
    Catalog Number:
    100213-RP02
    Price:
    None
    Category:
    Primary Antibody
    Reactivity:
    MERS CoV
    Applications:
    WB
    Immunogen:
    A synthetic peptide corresponding to the center region of MERS-CoV (NCoV / Novel coronavirus) Nucleocapsid Protein (NP protein).
    Product Aliases:
    Anti-coronavirus NP Antibody, Anti-coronavirus Nucleocapsid Antibody, Anti-coronavirus Nucleoprotein Antibody, Anti-cov np Antibody, Anti-ncov NP Antibody, Anti-novel coronavirus Nucleoprotein Antibody, Anti-NP Antibody, Anti-Nucleocapsid Antibody, Anti-Nucleoprotein Antibody
    Antibody Type:
    PAb
    Host:
    Rabbit
    Isotype:
    Rabbit IgG
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    Structured Review

    Sino Biological mers cov nucleocapsid protein antibody rabbit pab
    IHC detection of virus antigen expression in mouse tissue after challenge with <t>MERS-CoV.</t> Lung (A–C) and trachea (D–F) sections were assessed using rabbit polyclonal antibody to MERS-CoV nucleoprotein (NP) 3 days after the MERS-CoV challenge. The dark purple spot marked the inflammatory cell infiltration, and the brown particle marked the antigen of MERS-CoV. The MERS-CoV was located mainly in the trachea. Additionally, the lung tissue showed MERS-CoV expression in all immunized groups.
    Produced in rabbits immunized with purified a synthetic peptide corresponding to the center region of MERS CoV NCoV Novel coronavirus Nucleocapsid Protein NP protein and purified by antigen affinity chromatography
    https://www.bioz.com/result/mers cov nucleocapsid protein antibody rabbit pab/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mers cov nucleocapsid protein antibody rabbit pab - by Bioz Stars, 2021-07
    94/100 stars

    Images

    1) Product Images from "The recombinant N-terminal domain of spike proteins is a potential vaccine against Middle East respiratory syndrome coronavirus (MERS-CoV) infection"

    Article Title: The recombinant N-terminal domain of spike proteins is a potential vaccine against Middle East respiratory syndrome coronavirus (MERS-CoV) infection

    Journal: Vaccine

    doi: 10.1016/j.vaccine.2016.11.064

    IHC detection of virus antigen expression in mouse tissue after challenge with MERS-CoV. Lung (A–C) and trachea (D–F) sections were assessed using rabbit polyclonal antibody to MERS-CoV nucleoprotein (NP) 3 days after the MERS-CoV challenge. The dark purple spot marked the inflammatory cell infiltration, and the brown particle marked the antigen of MERS-CoV. The MERS-CoV was located mainly in the trachea. Additionally, the lung tissue showed MERS-CoV expression in all immunized groups.
    Figure Legend Snippet: IHC detection of virus antigen expression in mouse tissue after challenge with MERS-CoV. Lung (A–C) and trachea (D–F) sections were assessed using rabbit polyclonal antibody to MERS-CoV nucleoprotein (NP) 3 days after the MERS-CoV challenge. The dark purple spot marked the inflammatory cell infiltration, and the brown particle marked the antigen of MERS-CoV. The MERS-CoV was located mainly in the trachea. Additionally, the lung tissue showed MERS-CoV expression in all immunized groups.

    Techniques Used: Immunohistochemistry, Expressing

    rNTD or rRBD vaccination reduced respiratory tract pathology in mice after MERS-CoV challenge. Representative results of hematoxylin-eosin (HE) staining in the lung (A – C) and trachea (D – F) of mock-treated or immunized mice. Severe lesions including the loss of pulmonary alveolus (represented by the white vacuole) and diffuse inflammatory cell infiltration (represented by the dark purple point) are shown (figure A). In contrast, milder lesions were observed among mice immunized with rRBD (figure B) or NTD (figure C), as the pulmonary alveolus was highly visible with less inflammatory cell infiltration. Inflammatory cell infiltration and impaired epithelium of the tunica mucosa bronchiorum were seen in the mock group (D). rRBD (E) or rNTD (F) alleviated the pathologic damage in the trachea of immunized mice. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)
    Figure Legend Snippet: rNTD or rRBD vaccination reduced respiratory tract pathology in mice after MERS-CoV challenge. Representative results of hematoxylin-eosin (HE) staining in the lung (A – C) and trachea (D – F) of mock-treated or immunized mice. Severe lesions including the loss of pulmonary alveolus (represented by the white vacuole) and diffuse inflammatory cell infiltration (represented by the dark purple point) are shown (figure A). In contrast, milder lesions were observed among mice immunized with rRBD (figure B) or NTD (figure C), as the pulmonary alveolus was highly visible with less inflammatory cell infiltration. Inflammatory cell infiltration and impaired epithelium of the tunica mucosa bronchiorum were seen in the mock group (D). rRBD (E) or rNTD (F) alleviated the pathologic damage in the trachea of immunized mice. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

    Techniques Used: Mouse Assay, Staining

    Description of the N-terminal domain (NTD) immunogen and vaccination schedule. (A) The location of the NTD protein on the Middle East respiratory syndrome coronavirus MERS-CoV spike (S) protein. The recombinant (r)NTD protein consists of 336 amino acid (aa) residues (18–353) of S protein. A gp67 signal peptide (SP) was added to the N terminus for expression of the rNTD protein. (B) Purified rNTD protein detected by SDS-PAGE (left) and Western blot (right). The purified rNTD protein was separated by a 10% SDS-PAGE and stained with 0.25% Coomassie brilliant blue. Anti-NTD polyclonal antibody and infrared ray-labeled secondary antibody were used for the Western blot assay. Lane 1: protein molecular weight marker; lane 2: purified rNTD protein. (C). Vaccination schedule and detection. Mice received three vaccinations consisting of 5 or 10 μg of rNTD protein combined with adjuvants at 4-week intervals. Sera were collected at the indicated times to analyze the humoral immune response. Six mice from each group were sacrificed 2 weeks after the last immunization. The spleens were harvested for enzyme-linked immunospot (ELISpot), intracellular cytokine staining (ICS), and cytometric bead array (CBA) assays. In parallel experiments, the remaining mice were challenged with MERS-CoV to detect the protective effect elicited by the rNTD protein. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)
    Figure Legend Snippet: Description of the N-terminal domain (NTD) immunogen and vaccination schedule. (A) The location of the NTD protein on the Middle East respiratory syndrome coronavirus MERS-CoV spike (S) protein. The recombinant (r)NTD protein consists of 336 amino acid (aa) residues (18–353) of S protein. A gp67 signal peptide (SP) was added to the N terminus for expression of the rNTD protein. (B) Purified rNTD protein detected by SDS-PAGE (left) and Western blot (right). The purified rNTD protein was separated by a 10% SDS-PAGE and stained with 0.25% Coomassie brilliant blue. Anti-NTD polyclonal antibody and infrared ray-labeled secondary antibody were used for the Western blot assay. Lane 1: protein molecular weight marker; lane 2: purified rNTD protein. (C). Vaccination schedule and detection. Mice received three vaccinations consisting of 5 or 10 μg of rNTD protein combined with adjuvants at 4-week intervals. Sera were collected at the indicated times to analyze the humoral immune response. Six mice from each group were sacrificed 2 weeks after the last immunization. The spleens were harvested for enzyme-linked immunospot (ELISpot), intracellular cytokine staining (ICS), and cytometric bead array (CBA) assays. In parallel experiments, the remaining mice were challenged with MERS-CoV to detect the protective effect elicited by the rNTD protein. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

    Techniques Used: Recombinant, Expressing, Purification, SDS Page, Western Blot, Staining, Labeling, Molecular Weight, Marker, Mouse Assay, Enzyme-linked Immunospot, Crocin Bleaching Assay

    2) Product Images from "Inhibiting coronavirus replication in cultured cells by chemical ER stress"

    Article Title: Inhibiting coronavirus replication in cultured cells by chemical ER stress

    Journal: bioRxiv

    doi: 10.1101/2020.08.26.266304

    Thapsigargin inhibits the replication of high- and low-pathogenic human coronaviruses in multiple cell types. (A-D) Human embryonic MRC-5 lung fibroblasts were infected with HCoV-229E according to the scheme shown in Fig. 1B . Viral titers (A, upper graph), and expression of viral S gene-containing RNAs (A, lower graph), viral and host cell proteins (B, C) and cell viability (D) were analyzed and quantified as described in the legend of Fig. 1 . (E-K) Similarly, HuH7 cells or Vero E6 African green monkey kidney epithelial cells were infected with MERS-CoV (MOI=0.5) or SARS-CoV-2 (MOI=0.5) for 12 h or 24 in the presence / absence of 0.4 μM or 1 μM thapsigargin. (E, F) show viral titers and (G, H) the corresponding expression of MERS-CoV / SARS-CoV-2 nucleocapsid (N) and host cell proteins, respectively. (I) Dose-dependent suppression of MERS-CoV-2 replication by thapsigargin (upper graph) and the estimated effective inhibitory concentration (EC 50 ) in HuH7 cells infected with an MOI of 0.5 (lower graph). (J) Dose-dependent suppression of SARS-CoV-2 replication by thapsigargin (left graph) and the calculated effective inhibitory concentration (EC 50 ) in Vero E6 cells infected with an MOI of 0.5 (right graph). (K) The CC 50 of thapsigargin in Vero E6 cells was calculated by MTS assays as described in the legend of Fig. 2G-H . Data points show values from independent biological replicates, error bars show s.d.. Asterisks indicate p values (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, **** p ≤ 0.0001) obtained by two-tailed unpaired t-tests. See Fig. S2 for quantifications from replicates for MERS-CoV / SARS-CoV-2 immunoblot experiments.
    Figure Legend Snippet: Thapsigargin inhibits the replication of high- and low-pathogenic human coronaviruses in multiple cell types. (A-D) Human embryonic MRC-5 lung fibroblasts were infected with HCoV-229E according to the scheme shown in Fig. 1B . Viral titers (A, upper graph), and expression of viral S gene-containing RNAs (A, lower graph), viral and host cell proteins (B, C) and cell viability (D) were analyzed and quantified as described in the legend of Fig. 1 . (E-K) Similarly, HuH7 cells or Vero E6 African green monkey kidney epithelial cells were infected with MERS-CoV (MOI=0.5) or SARS-CoV-2 (MOI=0.5) for 12 h or 24 in the presence / absence of 0.4 μM or 1 μM thapsigargin. (E, F) show viral titers and (G, H) the corresponding expression of MERS-CoV / SARS-CoV-2 nucleocapsid (N) and host cell proteins, respectively. (I) Dose-dependent suppression of MERS-CoV-2 replication by thapsigargin (upper graph) and the estimated effective inhibitory concentration (EC 50 ) in HuH7 cells infected with an MOI of 0.5 (lower graph). (J) Dose-dependent suppression of SARS-CoV-2 replication by thapsigargin (left graph) and the calculated effective inhibitory concentration (EC 50 ) in Vero E6 cells infected with an MOI of 0.5 (right graph). (K) The CC 50 of thapsigargin in Vero E6 cells was calculated by MTS assays as described in the legend of Fig. 2G-H . Data points show values from independent biological replicates, error bars show s.d.. Asterisks indicate p values (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, **** p ≤ 0.0001) obtained by two-tailed unpaired t-tests. See Fig. S2 for quantifications from replicates for MERS-CoV / SARS-CoV-2 immunoblot experiments.

    Techniques Used: Infection, Expressing, Concentration Assay, Two Tailed Test

    Proteome profiling of thapsigargin effects on MERS-CoV or SARS-CoV-2-infected cells reveals multiple virus- and thapsigargin-specific protein and pathway patterns. (A-D) Total cell extracts from uninfected cells (-), HuH7 cells infected with MERS-CoV (M, MOI=0.5) for 12 h (A) or 24 h (B), or Vero E6 cells infected with SARS-CoV-2 (S, MOI=0.5) for 12 h (C) or 24 h (D), in the presence or absence of thapsigargin (T, 1 μM) were subjected to LC-MS/MS analysis. In total, 5,372 (from HuH7) or 5,072 (from Vero E6 cells) majority protein IDs were identified. Quantile-normalized log 2 -tranformed protein intensities were used for all further calculations. Volcano plots show the distribution of pairwise ratio comparisons and the corresponding p values which were obtained by pooling data from two independent experiments and from three technical replicates per sample. Blue and red colors indicate differentially expressed proteins (DEPs, ratio > 0, p ≥ −log 10 1.3). Purple and light red colors indicate individual viral proteins. (E) Scheme of multiple gene ID lists (corresponding to the DEPs shown in (A)-(D)) that were used for overrepresentation analyses to identify the top 100 enriched pathway categories per virus and time point using Metascape software ( Zhou et al ., 2019 ). Complete lists of pathways are shown in Fig. S3 and S4 as clustered heatmaps. The top 5 enriched pathway categories for up- or downregulated DEPs are shown in Fig. S5A . (F, G) The 400 enriched pathway categories were pooled and filtered for common and distinct pathways considering only terms with enrichment p values ≤ log 10 −3. (F) Venn diagram showing pathway terms specific to MERS-CoV (M), SARS-CoV-2 (S) or thapsigargin (T). The top 20 enriched pathway categories are shown in Fig. S5B . (G) Venn diagram showing pathway terms specific for virus, thapsigargin, or infection plus thapsigargin (virus + T) conditions. (H) The heatmap shows the top differentially enriched pathways corresponding to the Venn diagram shown in (G). Green colors refer to the pathways highlighted in (I). (I) Ten examples of differential and joint gene ID compositions of pathways enriched in HuH7 or Vero E6 cells.
    Figure Legend Snippet: Proteome profiling of thapsigargin effects on MERS-CoV or SARS-CoV-2-infected cells reveals multiple virus- and thapsigargin-specific protein and pathway patterns. (A-D) Total cell extracts from uninfected cells (-), HuH7 cells infected with MERS-CoV (M, MOI=0.5) for 12 h (A) or 24 h (B), or Vero E6 cells infected with SARS-CoV-2 (S, MOI=0.5) for 12 h (C) or 24 h (D), in the presence or absence of thapsigargin (T, 1 μM) were subjected to LC-MS/MS analysis. In total, 5,372 (from HuH7) or 5,072 (from Vero E6 cells) majority protein IDs were identified. Quantile-normalized log 2 -tranformed protein intensities were used for all further calculations. Volcano plots show the distribution of pairwise ratio comparisons and the corresponding p values which were obtained by pooling data from two independent experiments and from three technical replicates per sample. Blue and red colors indicate differentially expressed proteins (DEPs, ratio > 0, p ≥ −log 10 1.3). Purple and light red colors indicate individual viral proteins. (E) Scheme of multiple gene ID lists (corresponding to the DEPs shown in (A)-(D)) that were used for overrepresentation analyses to identify the top 100 enriched pathway categories per virus and time point using Metascape software ( Zhou et al ., 2019 ). Complete lists of pathways are shown in Fig. S3 and S4 as clustered heatmaps. The top 5 enriched pathway categories for up- or downregulated DEPs are shown in Fig. S5A . (F, G) The 400 enriched pathway categories were pooled and filtered for common and distinct pathways considering only terms with enrichment p values ≤ log 10 −3. (F) Venn diagram showing pathway terms specific to MERS-CoV (M), SARS-CoV-2 (S) or thapsigargin (T). The top 20 enriched pathway categories are shown in Fig. S5B . (G) Venn diagram showing pathway terms specific for virus, thapsigargin, or infection plus thapsigargin (virus + T) conditions. (H) The heatmap shows the top differentially enriched pathways corresponding to the Venn diagram shown in (G). Green colors refer to the pathways highlighted in (I). (I) Ten examples of differential and joint gene ID compositions of pathways enriched in HuH7 or Vero E6 cells.

    Techniques Used: Infection, Liquid Chromatography with Mass Spectroscopy, Software

    Top pathways regulated by MERS-CoV, SARS-CoV-2 or thapsigargin. (A) Top ten enriched pathways containing up- or downregulated DEPs extracted from the 100 enriched deregulated pathways shown in Fig. S3 / Fig. S4 . Colors indicate highly common categories. (B) Top 20 pathways enriched with thapsigargin alone or jointly by MERS-CoV, SARS-CoV-2 and thapsigargin according to the Venn diagram shown in Fig. 4F .
    Figure Legend Snippet: Top pathways regulated by MERS-CoV, SARS-CoV-2 or thapsigargin. (A) Top ten enriched pathways containing up- or downregulated DEPs extracted from the 100 enriched deregulated pathways shown in Fig. S3 / Fig. S4 . Colors indicate highly common categories. (B) Top 20 pathways enriched with thapsigargin alone or jointly by MERS-CoV, SARS-CoV-2 and thapsigargin according to the Venn diagram shown in Fig. 4F .

    Techniques Used:

    Opposing effects of HCoV-229E or MERS-CoV on ER stress genes at the mRNA and protein level. Projection of normalized ratio values for transcriptomic (by RNA-seq) and proteomic (by LC-MS/MS) data derived in parallel from HuH7 cells infected for 24 h with HCoV-229E or MERS-CoV with a MOI=1 on the components of the KEGG pathway 04141 “protein processing in endoplasmic reticulum”. The left side of the boxes show mRNA values, right sides show protein values.
    Figure Legend Snippet: Opposing effects of HCoV-229E or MERS-CoV on ER stress genes at the mRNA and protein level. Projection of normalized ratio values for transcriptomic (by RNA-seq) and proteomic (by LC-MS/MS) data derived in parallel from HuH7 cells infected for 24 h with HCoV-229E or MERS-CoV with a MOI=1 on the components of the KEGG pathway 04141 “protein processing in endoplasmic reticulum”. The left side of the boxes show mRNA values, right sides show protein values.

    Techniques Used: RNA Sequencing Assay, Liquid Chromatography with Mass Spectroscopy, Derivative Assay, Infection

    Thapsigargin suppresses MERS-CoV and SARS-CoV-2 N protein and upregulates BiP in infected cells. (A, B) show the quantification of replicate immunoblot experiments performed as shown in Fig. 3G and 3H . Data points show values from independent biological replicates, error bars show s.d..
    Figure Legend Snippet: Thapsigargin suppresses MERS-CoV and SARS-CoV-2 N protein and upregulates BiP in infected cells. (A, B) show the quantification of replicate immunoblot experiments performed as shown in Fig. 3G and 3H . Data points show values from independent biological replicates, error bars show s.d..

    Techniques Used: Infection

    Thapsigargin effects on MERS-CoV or SARS-CoV-2-infected cells reveal the regulation of a specific network of proteins involved in transport, ERQC / ERAD and ER stress. (A) Venn diagram demonstrating the overlap of orthologues proteins expressed in HuH7 and Ve ro cells based on the NCBI gene IDs corresponding to majority protein IDs. (B) The overlap of virus- and thapsigargin-regulated proteins common to HuH7 and Vero E6 cells was calculated based on gene IDs. This analysis identifies 180 proteins with higher and 61 proteins with lower expression in thapsigargin-treated infected cells compared to virus infection alone (ratio > 0, p ≥ −log 10 1.3). (C) Heatmaps showing individual mean ratio values of log 2 -transfomed normalized protein intensities for the top 50 up- or downregulated proteins in virus-infected and thapsigargin-treated cells. Ratio values of infected or thapsigargin-treated conditions compared to untreated cells (-) cells are shown for comparison. Note that ratios are sorted and color coded according to the virus plus thapsigargin conditions with infection alone conditions set as denominator. Green colors highlight HERPUD1 and BiP (HSPA5) values. Orange colors highlight SQSMT1 which was also identified as SARS-CoV-2-regulated protein in an independent study ( Stukalov et al ., 2020 ). (D) Overrepresentation analysis showing the top 10 pathways mapping to gene IDs with increased (180 proteins, red) or decreased (61 proteins, blue) expression levels in thapsigargin-treated and infected cells compared to virus infection alone. Gene ID lists were analyzed using Metascape software ( Zhou et al ., 2019 ). (E) Experimental evidence, co-occurrence, co-expression and confidence scores from the STRING database ( Szklarczyk et al , 2019 ) were used to identify protein:protein interactions (PPI) amongst the 180 up- and 61 downregulated thapsigargin-sensitive proteins. As shown, based on experimental evidence and combined STRING score criteria, 59 and 26 coregulated proteins are engaged in defined PPI networks; the remaining proteins are not known to interact. (F) Validation of thapsigargin-induced HERPUD1 and CTH upregulation by immunoblotting of HuH7 or Vero E6 whole cell extracts from cells treated as described above. BiP and IRE1α levels are shown for comparison. (G) Quantification of thapsigargin-mediated re-expression of HERPUD1 and CTH in cells infected with HCoV-229E, MERS-CoV or SARS-CoV-2 from two independent immunoblot experiments. Error bars show s.d.. (H) Heatmap showing thapsigargin-reprogrammed proteins of KEGG 04141 (mean ratio ≥ 1.5 fold) along with p values. See also Fig. S6 for projection of thapsigargin-mediated protein changes on the KEGG pathway map. (I) Venn diagram showing the intersection of thapsigargin- / virus-regulated proteins with all novel ERAD components (FDR of 1 %) identified by ( Leto et al ., 2019 ). The regulation of 31 overlapping components is shown as a heatmap displaying mean ratio values in thapsigargin-treated or infected cells. Red colors highlight UBA5 and ZNF622 as discussed in the text. (J) Summary of the main findings of our study. Abbreviations: M, MERS-CoV; S, SARS-CoV-2; T, thapsigargin.
    Figure Legend Snippet: Thapsigargin effects on MERS-CoV or SARS-CoV-2-infected cells reveal the regulation of a specific network of proteins involved in transport, ERQC / ERAD and ER stress. (A) Venn diagram demonstrating the overlap of orthologues proteins expressed in HuH7 and Ve ro cells based on the NCBI gene IDs corresponding to majority protein IDs. (B) The overlap of virus- and thapsigargin-regulated proteins common to HuH7 and Vero E6 cells was calculated based on gene IDs. This analysis identifies 180 proteins with higher and 61 proteins with lower expression in thapsigargin-treated infected cells compared to virus infection alone (ratio > 0, p ≥ −log 10 1.3). (C) Heatmaps showing individual mean ratio values of log 2 -transfomed normalized protein intensities for the top 50 up- or downregulated proteins in virus-infected and thapsigargin-treated cells. Ratio values of infected or thapsigargin-treated conditions compared to untreated cells (-) cells are shown for comparison. Note that ratios are sorted and color coded according to the virus plus thapsigargin conditions with infection alone conditions set as denominator. Green colors highlight HERPUD1 and BiP (HSPA5) values. Orange colors highlight SQSMT1 which was also identified as SARS-CoV-2-regulated protein in an independent study ( Stukalov et al ., 2020 ). (D) Overrepresentation analysis showing the top 10 pathways mapping to gene IDs with increased (180 proteins, red) or decreased (61 proteins, blue) expression levels in thapsigargin-treated and infected cells compared to virus infection alone. Gene ID lists were analyzed using Metascape software ( Zhou et al ., 2019 ). (E) Experimental evidence, co-occurrence, co-expression and confidence scores from the STRING database ( Szklarczyk et al , 2019 ) were used to identify protein:protein interactions (PPI) amongst the 180 up- and 61 downregulated thapsigargin-sensitive proteins. As shown, based on experimental evidence and combined STRING score criteria, 59 and 26 coregulated proteins are engaged in defined PPI networks; the remaining proteins are not known to interact. (F) Validation of thapsigargin-induced HERPUD1 and CTH upregulation by immunoblotting of HuH7 or Vero E6 whole cell extracts from cells treated as described above. BiP and IRE1α levels are shown for comparison. (G) Quantification of thapsigargin-mediated re-expression of HERPUD1 and CTH in cells infected with HCoV-229E, MERS-CoV or SARS-CoV-2 from two independent immunoblot experiments. Error bars show s.d.. (H) Heatmap showing thapsigargin-reprogrammed proteins of KEGG 04141 (mean ratio ≥ 1.5 fold) along with p values. See also Fig. S6 for projection of thapsigargin-mediated protein changes on the KEGG pathway map. (I) Venn diagram showing the intersection of thapsigargin- / virus-regulated proteins with all novel ERAD components (FDR of 1 %) identified by ( Leto et al ., 2019 ). The regulation of 31 overlapping components is shown as a heatmap displaying mean ratio values in thapsigargin-treated or infected cells. Red colors highlight UBA5 and ZNF622 as discussed in the text. (J) Summary of the main findings of our study. Abbreviations: M, MERS-CoV; S, SARS-CoV-2; T, thapsigargin.

    Techniques Used: Infection, Expressing, Software

    CoVs uncouple mRNA and protein levels of ER stress components in infected cells. HuH7 cells were left untreated or were infected with HCoV-229E or MERS-CoV (MOI=1) for 3, 6, 12, or 24 h. Transcriptomic data (by RNA-seq, n=2) and proteomic data (by LC-MS/MS, n=2, three technical replicates per sample) were derived from samples obtained at the indicated time points p.i. and subsequently used to extract expression values for the KEGG pathway 04141 “protein processing in endoplasmic reticulum”. (A) Scatter plots show mean normalized protein / mRNA expression values for each component, fitted linear regression lines, confidence intervals and correlation coefficients for mock-infected HuH7 cells and HuH7 cells infected for 24 h. (B) Correlation matrix of Pearson’s r across all conditions. All p values are given in supplementary Table S1. (C) The heatmap shows mean ratio values of differentially expressed mRNAs or proteins based on significant differences (fold change ≥ 2, p ≤ 0.01) calculated from the two biological replicates. See also Fig. S1 and Table S1.
    Figure Legend Snippet: CoVs uncouple mRNA and protein levels of ER stress components in infected cells. HuH7 cells were left untreated or were infected with HCoV-229E or MERS-CoV (MOI=1) for 3, 6, 12, or 24 h. Transcriptomic data (by RNA-seq, n=2) and proteomic data (by LC-MS/MS, n=2, three technical replicates per sample) were derived from samples obtained at the indicated time points p.i. and subsequently used to extract expression values for the KEGG pathway 04141 “protein processing in endoplasmic reticulum”. (A) Scatter plots show mean normalized protein / mRNA expression values for each component, fitted linear regression lines, confidence intervals and correlation coefficients for mock-infected HuH7 cells and HuH7 cells infected for 24 h. (B) Correlation matrix of Pearson’s r across all conditions. All p values are given in supplementary Table S1. (C) The heatmap shows mean ratio values of differentially expressed mRNAs or proteins based on significant differences (fold change ≥ 2, p ≤ 0.01) calculated from the two biological replicates. See also Fig. S1 and Table S1.

    Techniques Used: Infection, RNA Sequencing Assay, Liquid Chromatography with Mass Spectroscopy, Derivative Assay, Expressing

    Identification of deregulated cellular pathways in MERS-CoV-infected HuH7 cells. Top 100 overrepresented pathways containing up- or downregulated DEPs (ratio > 0, p ≥ log 10 1.3) for the 12 h p.i. and 24 h p.i. time points of MERS-CoV-infected cells based on gene IDs derived from protein IDs. Blue and red colors indicate differentially expressed proteins as shown in Fig. 4A-B .
    Figure Legend Snippet: Identification of deregulated cellular pathways in MERS-CoV-infected HuH7 cells. Top 100 overrepresented pathways containing up- or downregulated DEPs (ratio > 0, p ≥ log 10 1.3) for the 12 h p.i. and 24 h p.i. time points of MERS-CoV-infected cells based on gene IDs derived from protein IDs. Blue and red colors indicate differentially expressed proteins as shown in Fig. 4A-B .

    Techniques Used: Infection, Derivative Assay

    3) Product Images from "Recombinant Receptor Binding Domain Protein Induces Partial Protective Immunity in Rhesus Macaques Against Middle East Respiratory Syndrome Coronavirus Challenge"

    Article Title: Recombinant Receptor Binding Domain Protein Induces Partial Protective Immunity in Rhesus Macaques Against Middle East Respiratory Syndrome Coronavirus Challenge

    Journal: EBioMedicine

    doi: 10.1016/j.ebiom.2015.08.031

    MERS-CoV loads in the swabs and tissues of immunised rhesus macaques following MERS-CoV infection detected by Real-time-PCR. a. Viral loads in swab samples at 1 dpi and 3 dpi. b. The viral loads in lung and trachea tissue 3 days after MERS-CoV infection. All of the detections were replicated for three times. M, H and L represent mock, high- and low-dose groups, respectively. * p
    Figure Legend Snippet: MERS-CoV loads in the swabs and tissues of immunised rhesus macaques following MERS-CoV infection detected by Real-time-PCR. a. Viral loads in swab samples at 1 dpi and 3 dpi. b. The viral loads in lung and trachea tissue 3 days after MERS-CoV infection. All of the detections were replicated for three times. M, H and L represent mock, high- and low-dose groups, respectively. * p

    Techniques Used: Infection, Real-time Polymerase Chain Reaction

    IHC staining of immunised rhesus macaque lung and trachea 3 days after MERS-CoV infection by a polyclonal antibody against nucleoprotein. a–c. IHC analysis of lung tissue. d–f. IHC staining of trachea tissue. Black arrows indicate the distribution of viral antigen. M, H and L represent mock, high- and low-dose groups, respectively.
    Figure Legend Snippet: IHC staining of immunised rhesus macaque lung and trachea 3 days after MERS-CoV infection by a polyclonal antibody against nucleoprotein. a–c. IHC analysis of lung tissue. d–f. IHC staining of trachea tissue. Black arrows indicate the distribution of viral antigen. M, H and L represent mock, high- and low-dose groups, respectively.

    Techniques Used: Immunohistochemistry, Staining, Infection

    Schematic diagram of the rRBD vaccination and MERS-CoV challenge schedule. Monkeys were immunised three times intramuscularly (i.m.), at intervals of 8 or 17 weeks, and inoculated intratracheally with hCoV-EMC at a dosage of 6.5 × 10 7 TCID 50 (red arrow). Monkeys were bled periodically at 2, 10, and 25 weeks (− 14 days), and at − 1 day, 1 day and 3 days. Additionally, nasal, oral and rectal swabs were collected at − 1 day, 1 day and 3 days. Chest X-rays were performed 1 day before inoculation, and at 1 and 3 dpi. Following euthanasia, lung, trachea, spleen and kidney specimens of challenged monkeys were acquired at 3 dpi for detection.
    Figure Legend Snippet: Schematic diagram of the rRBD vaccination and MERS-CoV challenge schedule. Monkeys were immunised three times intramuscularly (i.m.), at intervals of 8 or 17 weeks, and inoculated intratracheally with hCoV-EMC at a dosage of 6.5 × 10 7 TCID 50 (red arrow). Monkeys were bled periodically at 2, 10, and 25 weeks (− 14 days), and at − 1 day, 1 day and 3 days. Additionally, nasal, oral and rectal swabs were collected at − 1 day, 1 day and 3 days. Chest X-rays were performed 1 day before inoculation, and at 1 and 3 dpi. Following euthanasia, lung, trachea, spleen and kidney specimens of challenged monkeys were acquired at 3 dpi for detection.

    Techniques Used:

    Radiographic alterations and lung pathology. a. X-rays from rhesus macaques imaged prior to (− 1 day) and post-MERS-CoV inoculation (1 day and 3 days). Areas of interstitial infiltration, indicative of pneumonia, are highlighted (circle). b. Gross pathology of the lungs from necropsied animals at 3 dpi. Haemorrhage and necrosis indicated by the black circle in both high and low dose immunisation groups were small and local, which happened in the mock group were large and disperse. M, H and L represent the mock, high- and low-dose groups, respectively.
    Figure Legend Snippet: Radiographic alterations and lung pathology. a. X-rays from rhesus macaques imaged prior to (− 1 day) and post-MERS-CoV inoculation (1 day and 3 days). Areas of interstitial infiltration, indicative of pneumonia, are highlighted (circle). b. Gross pathology of the lungs from necropsied animals at 3 dpi. Haemorrhage and necrosis indicated by the black circle in both high and low dose immunisation groups were small and local, which happened in the mock group were large and disperse. M, H and L represent the mock, high- and low-dose groups, respectively.

    Techniques Used:

    Histopathological changes in the lungs and tracheas of immunised rhesus macaques following MERS-CoV challenge. Three days after MERS-CoV inoculation, all monkeys were euthanised. Tissues were collected and stained with haematoxylin and eosin (H E). a–f. Pathological changes in the lungs of immunised monkeys. The black triangle indicates acute interstitial pneumonia diffused in lung tissue. The unfilled arrow indicates the hyaline member resulting from effusion of fibrin. The black arrow indicates the influx of inflammatory cells. g–i. Pathological findings of tracheas in immunised rhesus macaques. The black arrow highlights the infiltration of inflammatory cells in the tunica mucosa bronchiorum. The unfilled arrow indicates epithelium impairment in tunica mucosa bronchiorum. M, H and L represent the mock, high- and low-dose groups, respectively.
    Figure Legend Snippet: Histopathological changes in the lungs and tracheas of immunised rhesus macaques following MERS-CoV challenge. Three days after MERS-CoV inoculation, all monkeys were euthanised. Tissues were collected and stained with haematoxylin and eosin (H E). a–f. Pathological changes in the lungs of immunised monkeys. The black triangle indicates acute interstitial pneumonia diffused in lung tissue. The unfilled arrow indicates the hyaline member resulting from effusion of fibrin. The black arrow indicates the influx of inflammatory cells. g–i. Pathological findings of tracheas in immunised rhesus macaques. The black arrow highlights the infiltration of inflammatory cells in the tunica mucosa bronchiorum. The unfilled arrow indicates epithelium impairment in tunica mucosa bronchiorum. M, H and L represent the mock, high- and low-dose groups, respectively.

    Techniques Used: Staining

    4) Product Images from "Enhanced protection in mice induced by immunization with inactivated whole viruses compare to spike protein of middle east respiratory syndrome coronavirus"

    Article Title: Enhanced protection in mice induced by immunization with inactivated whole viruses compare to spike protein of middle east respiratory syndrome coronavirus

    Journal: Emerging Microbes & Infections

    doi: 10.1038/s41426-018-0056-7

    Neutralizing antibodies induced by S protein and IV vaccine against MERS-CoV pseudovirus particles and MERS-CoV. Neutralizing antibody titers against MERS-CoV pseudovirus particles. ( a , b ) and MERS-CoV ( c , d ) were determined by plaque reduction neutralization assays at 6 and 10 weeks. Representative results of the plaque reduction neutralization (PRNT) assay for the detection of neutralization activity in the sera of mice ( e ). Approximately 30 pfu of the virus stock (hCoV-EMC) was used to infect Vero cells in 12-well plates with or without heat-inactivated sera from immunized mice 2 weeks after the third immunization. PRNT50 was calculated after the plaques were counted. Significant values are defined by ** p
    Figure Legend Snippet: Neutralizing antibodies induced by S protein and IV vaccine against MERS-CoV pseudovirus particles and MERS-CoV. Neutralizing antibody titers against MERS-CoV pseudovirus particles. ( a , b ) and MERS-CoV ( c , d ) were determined by plaque reduction neutralization assays at 6 and 10 weeks. Representative results of the plaque reduction neutralization (PRNT) assay for the detection of neutralization activity in the sera of mice ( e ). Approximately 30 pfu of the virus stock (hCoV-EMC) was used to infect Vero cells in 12-well plates with or without heat-inactivated sera from immunized mice 2 weeks after the third immunization. PRNT50 was calculated after the plaques were counted. Significant values are defined by ** p

    Techniques Used: Neutralization, Plaque Reduction Neutralization Test, Activity Assay, Mouse Assay

    The IV formulation provides enhanced protection in mice compared to the S formulation, indicating ameliorated lung pathology, reduced viral titers and expression of virus antigens in the lungs of mice with IV or S. Representative results of hematoxylin-eosin (HE) staining (×400) in the lungs of mock-treated or immunized mice ( a ). Immunohistochemistry staining (×400) with anti-S and anti-NP mAbs ( b ). Lung virus titers 3 days after MERS-CoV challenge ( c ) as detected by virus isolation and titration at day 3 post-challenge. Values are the means ± SEM. Significant values are defined by *** P
    Figure Legend Snippet: The IV formulation provides enhanced protection in mice compared to the S formulation, indicating ameliorated lung pathology, reduced viral titers and expression of virus antigens in the lungs of mice with IV or S. Representative results of hematoxylin-eosin (HE) staining (×400) in the lungs of mock-treated or immunized mice ( a ). Immunohistochemistry staining (×400) with anti-S and anti-NP mAbs ( b ). Lung virus titers 3 days after MERS-CoV challenge ( c ) as detected by virus isolation and titration at day 3 post-challenge. Values are the means ± SEM. Significant values are defined by *** P

    Techniques Used: Mouse Assay, Expressing, Staining, Immunohistochemistry, Virus Isolation Assay, Titration

    Related Articles

    Incubation:

    Article Title: The recombinant N-terminal domain of spike proteins is a potential vaccine against Middle East respiratory syndrome coronavirus (MERS-CoV) infection
    Article Snippet: For IHC analysis, staining methods included antigen retrieval with citrate buffer. .. Rabbit-serum-derived polyclonal antibody against nucleoprotein (cat: 100213-RP02; Sino Biological Inc., Beijing, CHN) was incubated with the sections at 1:1000 dilution; goat anti-rabbit (cat: pv-9001; ZSGB-Bio, Beijing, CHN) secondary antibody was used at 1:2000, and sections were evaluated using light microscopy. .. Various sites of the lungs and trachea tissue samples were collected for histopathology and IHC analyses.

    Article Title: Recombinant Receptor Binding Domain Protein Induces Partial Protective Immunity in Rhesus Macaques Against Middle East Respiratory Syndrome Coronavirus Challenge
    Article Snippet: For immunohistochemistry analysis, staining methods included antigen retrieval with citrate buffer. .. A rabbit-serum-derived polyclonal antibody against nucleoprotein (Sino Biological Inc., cat: 100213-RP02) at 1:1000 dilution, was then incubated with the sections. .. The secondary antibody used was goat anti-rabbit (ZSGB-Bio, cat: pv-9001) at 1:2000.

    Light Microscopy:

    Article Title: The recombinant N-terminal domain of spike proteins is a potential vaccine against Middle East respiratory syndrome coronavirus (MERS-CoV) infection
    Article Snippet: For IHC analysis, staining methods included antigen retrieval with citrate buffer. .. Rabbit-serum-derived polyclonal antibody against nucleoprotein (cat: 100213-RP02; Sino Biological Inc., Beijing, CHN) was incubated with the sections at 1:1000 dilution; goat anti-rabbit (cat: pv-9001; ZSGB-Bio, Beijing, CHN) secondary antibody was used at 1:2000, and sections were evaluated using light microscopy. .. Various sites of the lungs and trachea tissue samples were collected for histopathology and IHC analyses.

    Positive Control:

    Article Title: Detection of MERS-CoV antigen on formalin-fixed paraffin-embedded nasal tissue of alpacas by immunohistochemistry using human monoclonal antibodies directed against different epitopes of the spike protein
    Article Snippet: Sections were finally slightly counterstained with Mayer’s hematoxylin (Roth C. GmbH & Co KG). .. For positive control and elucidation of virus distribution, additional sections were stained with two different commercially available, previously published monoclonal mouse and polyclonal rabbit anti-MERS-CoV nucleocapsid antibodies (Sino Biological Inc.; ; ), respectively. .. For negative controls, the primary antibody was replaced by ascites fluid from Balb/c mice (1:1,000) and normal rabbit serum (1:3,000), respectively.

    Staining:

    Article Title: Detection of MERS-CoV antigen on formalin-fixed paraffin-embedded nasal tissue of alpacas by immunohistochemistry using human monoclonal antibodies directed against different epitopes of the spike protein
    Article Snippet: Sections were finally slightly counterstained with Mayer’s hematoxylin (Roth C. GmbH & Co KG). .. For positive control and elucidation of virus distribution, additional sections were stained with two different commercially available, previously published monoclonal mouse and polyclonal rabbit anti-MERS-CoV nucleocapsid antibodies (Sino Biological Inc.; ; ), respectively. .. For negative controls, the primary antibody was replaced by ascites fluid from Balb/c mice (1:1,000) and normal rabbit serum (1:3,000), respectively.

    other:

    Article Title: Saracatinib Inhibits Middle East Respiratory Syndrome-Coronavirus Replication In Vitro
    Article Snippet: Rabbit anti-MERS-CoV nucleocapsid (N) antibody was purchased from Sino Biological Inc. (Cat. 100211-RP02, Beijing, China).

    Article Title: Inhibiting coronavirus replication in cultured cells by chemical ER stress
    Article Snippet: Primary antibodies against the following proteins or peptides were used: anti β-actin (Santa Cruz, #sc-4778), anti PERK (Santa Cruz, #sc-377400), anti PERK (Abcam, #ab65142), anti BiP (Cell Signaling, #3177), anti eIF2α (Cell Signaling #9722), anti P(S51)-eIF2α (Cell Signaling #9721), anti P(S724)-IRE1α (Novus Biologicals, #NB100-2323), anti IRE1α (Santa Cruz, #sc-390960), anti ATF4 (Santa Cruz, #sc-390063), anti ATF3 (Santa Cruz, #sc-188), anti HERPUD1 antibody (Abnova, #H00009709-A01), anti CTH antibody (Cruz, #sc-374249), anti HCoV-229E N protein ((Ingenasa, Batch 250609), mouse anti HCoV-229E nsp12 (gift from Carsten Grötzinger), rabbit anti HCoV-229E nsp8 , anti MERS-CoV N protein (Sinobiological, #100213-RP02), rabbit anti SARS-CoV N protein cross-reacting with SARS-CoV-2 N protein (gift from Friedemann Weber), anti SARS-CoV-2 N protein (Rockland, #200-401-A50), anti puromycin (Kerafast Inc., 3RH11, #EQ 0001), anti J2 (SCICONS, English & Scientific Consulting Kft, #10010200).

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    Sino Biological mers cov nucleocapsid protein rabbit antibody
    Single-dose vaccination with ChAdOx1 <t>MERS</t> protects rhesus macaques against bronchointerstitial pneumonia caused by <t>MERS-CoV</t> challenge. Rhesus macaques were vaccinated with a prime-boost or prime-only regimen of ChAdOx1 MERS, or with ChAdOx1 GFP and challenged with MERS-CoV. (A) Ventrodorsal thoracic radiographs collected on 6 DPI. A marker (R) indicates right side of animal. No pathologic changes were observed in animals vaccinated with ChAdOx1 MERS via a prime-boost or prime-only regimen. Animal vaccinated with ChAdOx1 GFP shows focally extensive area of increased pulmonary opacity and deviation of the cardiac silhouette, highlighted in the circle located in the middle and caudal lung lobes. (B) Thoracic radiographs of each animal were scored per lung lobe resulting in a maximum score of 18. Values were averaged per group per day, shown is mean with standard deviation. See Table S2 for more details. (C) Gross pathology of lungs shows no pathologic changes in ChAdOx1 MERS vaccinated animals, and focally extensive areas of consolidation in left cranial, middle and caudal lung lobes in control animals (asterisks). (D) Gross lung lesions were scored for each lung lobe, ventral and dorsal. Values were averaged per group, shown is mean with standard deviation. (E) Lung tissue sections were stained with hematoxylin and eosin. Moderate numbers of lymphocyte accumulation around pulmonary arterioles (asterisks), and mild thickening of alveolar septae by lymphocytes and macrophages (arrow) in lung tissue of animals vaccinated with ChAdOx1 MERS. Marked bronchointerstitial pneumonia with abundant pulmonary edema and fibrin (asterisks), type II pneumocyte hyperplasia (arrow), and increased numbers of alveolar macrophages (arrowhead) in lung tissue of control animals. Magnification = 200x. (F) Lung to body weight ratio was determined for all animals at necropsy. Shown is mean with standard deviation. (G) Lung tissue sections were stained with antibody against MERS-CoV antigen, which is visible as a red-brown staining. No immunoreactivity was found in ChAdOx1 MERS vaccinated animals, whereas multifocal immunoreactivity of type I and II pneumocytes could be found in lung tissue of ChAdOx1 GFP vaccinated animals. (H) Lung tissue sections were scored on severity of lesions (0=no lesions, 1=1-10%, 2=11-25%, 3=26-50%, 4=51-75%, 5=76-100%) and averaged per group. Shown is mean with standard deviation. A = bronchointerstitial pneumonia; B = type II pneumocyte hyperplasia; C = Hemorrhages, edema, fibrin deposits. Statistical significance between groups was determined via two-tailed unpaired Student’s t-test. * = p-value
    Mers Cov Nucleocapsid Protein Rabbit Antibody, supplied by Sino Biological, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Sino Biological mers cov nucleoprotein np antibody rabbit pab
    Kinetics of serum and egg yolk <t>anti-MERS</t> <t>COV-S</t> IgY antibodies response of chickens after immunization with MERS COV-S recombinant protein compared with the adjuvant-immunized chicken (adjuvant control). Each week is represented by a pool of egg yolks of individual chicken in each group (S1-immunized and adjuvant-immunized).
    Mers Cov Nucleoprotein Np Antibody Rabbit Pab, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Single-dose vaccination with ChAdOx1 MERS protects rhesus macaques against bronchointerstitial pneumonia caused by MERS-CoV challenge. Rhesus macaques were vaccinated with a prime-boost or prime-only regimen of ChAdOx1 MERS, or with ChAdOx1 GFP and challenged with MERS-CoV. (A) Ventrodorsal thoracic radiographs collected on 6 DPI. A marker (R) indicates right side of animal. No pathologic changes were observed in animals vaccinated with ChAdOx1 MERS via a prime-boost or prime-only regimen. Animal vaccinated with ChAdOx1 GFP shows focally extensive area of increased pulmonary opacity and deviation of the cardiac silhouette, highlighted in the circle located in the middle and caudal lung lobes. (B) Thoracic radiographs of each animal were scored per lung lobe resulting in a maximum score of 18. Values were averaged per group per day, shown is mean with standard deviation. See Table S2 for more details. (C) Gross pathology of lungs shows no pathologic changes in ChAdOx1 MERS vaccinated animals, and focally extensive areas of consolidation in left cranial, middle and caudal lung lobes in control animals (asterisks). (D) Gross lung lesions were scored for each lung lobe, ventral and dorsal. Values were averaged per group, shown is mean with standard deviation. (E) Lung tissue sections were stained with hematoxylin and eosin. Moderate numbers of lymphocyte accumulation around pulmonary arterioles (asterisks), and mild thickening of alveolar septae by lymphocytes and macrophages (arrow) in lung tissue of animals vaccinated with ChAdOx1 MERS. Marked bronchointerstitial pneumonia with abundant pulmonary edema and fibrin (asterisks), type II pneumocyte hyperplasia (arrow), and increased numbers of alveolar macrophages (arrowhead) in lung tissue of control animals. Magnification = 200x. (F) Lung to body weight ratio was determined for all animals at necropsy. Shown is mean with standard deviation. (G) Lung tissue sections were stained with antibody against MERS-CoV antigen, which is visible as a red-brown staining. No immunoreactivity was found in ChAdOx1 MERS vaccinated animals, whereas multifocal immunoreactivity of type I and II pneumocytes could be found in lung tissue of ChAdOx1 GFP vaccinated animals. (H) Lung tissue sections were scored on severity of lesions (0=no lesions, 1=1-10%, 2=11-25%, 3=26-50%, 4=51-75%, 5=76-100%) and averaged per group. Shown is mean with standard deviation. A = bronchointerstitial pneumonia; B = type II pneumocyte hyperplasia; C = Hemorrhages, edema, fibrin deposits. Statistical significance between groups was determined via two-tailed unpaired Student’s t-test. * = p-value

    Journal: bioRxiv

    Article Title: A single dose of ChAdOx1 MERS provides broad protective immunity against a variety of MERS-CoV strains

    doi: 10.1101/2020.04.13.036293

    Figure Lengend Snippet: Single-dose vaccination with ChAdOx1 MERS protects rhesus macaques against bronchointerstitial pneumonia caused by MERS-CoV challenge. Rhesus macaques were vaccinated with a prime-boost or prime-only regimen of ChAdOx1 MERS, or with ChAdOx1 GFP and challenged with MERS-CoV. (A) Ventrodorsal thoracic radiographs collected on 6 DPI. A marker (R) indicates right side of animal. No pathologic changes were observed in animals vaccinated with ChAdOx1 MERS via a prime-boost or prime-only regimen. Animal vaccinated with ChAdOx1 GFP shows focally extensive area of increased pulmonary opacity and deviation of the cardiac silhouette, highlighted in the circle located in the middle and caudal lung lobes. (B) Thoracic radiographs of each animal were scored per lung lobe resulting in a maximum score of 18. Values were averaged per group per day, shown is mean with standard deviation. See Table S2 for more details. (C) Gross pathology of lungs shows no pathologic changes in ChAdOx1 MERS vaccinated animals, and focally extensive areas of consolidation in left cranial, middle and caudal lung lobes in control animals (asterisks). (D) Gross lung lesions were scored for each lung lobe, ventral and dorsal. Values were averaged per group, shown is mean with standard deviation. (E) Lung tissue sections were stained with hematoxylin and eosin. Moderate numbers of lymphocyte accumulation around pulmonary arterioles (asterisks), and mild thickening of alveolar septae by lymphocytes and macrophages (arrow) in lung tissue of animals vaccinated with ChAdOx1 MERS. Marked bronchointerstitial pneumonia with abundant pulmonary edema and fibrin (asterisks), type II pneumocyte hyperplasia (arrow), and increased numbers of alveolar macrophages (arrowhead) in lung tissue of control animals. Magnification = 200x. (F) Lung to body weight ratio was determined for all animals at necropsy. Shown is mean with standard deviation. (G) Lung tissue sections were stained with antibody against MERS-CoV antigen, which is visible as a red-brown staining. No immunoreactivity was found in ChAdOx1 MERS vaccinated animals, whereas multifocal immunoreactivity of type I and II pneumocytes could be found in lung tissue of ChAdOx1 GFP vaccinated animals. (H) Lung tissue sections were scored on severity of lesions (0=no lesions, 1=1-10%, 2=11-25%, 3=26-50%, 4=51-75%, 5=76-100%) and averaged per group. Shown is mean with standard deviation. A = bronchointerstitial pneumonia; B = type II pneumocyte hyperplasia; C = Hemorrhages, edema, fibrin deposits. Statistical significance between groups was determined via two-tailed unpaired Student’s t-test. * = p-value

    Article Snippet: Specific anti-CoV immunoreactivity was detected using MERS-CoV nucleocapsid protein rabbit antibody (Sino Biological Inc.) at a 1:4000.

    Techniques: Marker, Standard Deviation, Staining, Two Tailed Test

    ChAdOx1 MERS provides cross-protection against different MERS-CoV strains in the mouse model. (A) Survival curves of ChAdOx1 MERS vaccinated (solid line) and ChAdOx1 GFP vaccinated (dashed line) hDPP4 mice challenged with MERS-CoV. (B) Infectious virus titers in lung tissue collected on 3 DPI from hDPP4 mice challenged with MERS-CoV. Shown is mean titer with standard deviation.

    Journal: bioRxiv

    Article Title: A single dose of ChAdOx1 MERS provides broad protective immunity against a variety of MERS-CoV strains

    doi: 10.1101/2020.04.13.036293

    Figure Lengend Snippet: ChAdOx1 MERS provides cross-protection against different MERS-CoV strains in the mouse model. (A) Survival curves of ChAdOx1 MERS vaccinated (solid line) and ChAdOx1 GFP vaccinated (dashed line) hDPP4 mice challenged with MERS-CoV. (B) Infectious virus titers in lung tissue collected on 3 DPI from hDPP4 mice challenged with MERS-CoV. Shown is mean titer with standard deviation.

    Article Snippet: Specific anti-CoV immunoreactivity was detected using MERS-CoV nucleocapsid protein rabbit antibody (Sino Biological Inc.) at a 1:4000.

    Techniques: Mouse Assay, Standard Deviation

    Maximum likelihood phylogeny of all published full-length MERS-CoV nucleotide sequences. Strains highlighted in green: human isolates used in this study; strains highlighted in blue: camel isolates used in this study; strain highlighted in red: ChAdOx1 MERS strain.

    Journal: bioRxiv

    Article Title: A single dose of ChAdOx1 MERS provides broad protective immunity against a variety of MERS-CoV strains

    doi: 10.1101/2020.04.13.036293

    Figure Lengend Snippet: Maximum likelihood phylogeny of all published full-length MERS-CoV nucleotide sequences. Strains highlighted in green: human isolates used in this study; strains highlighted in blue: camel isolates used in this study; strain highlighted in red: ChAdOx1 MERS strain.

    Article Snippet: Specific anti-CoV immunoreactivity was detected using MERS-CoV nucleocapsid protein rabbit antibody (Sino Biological Inc.) at a 1:4000.

    Techniques:

    Vaccination of rhesus macaques with ChAdOx1 MERS elicits a humoral immune response. Serum samples were collected from non-human primates at times of vaccination (−56 DPI and -28 DPI), 14 days later and at challenge. (A) Overview of experimental timeline. V-M = vaccination with ChAdOx1 MERS; V-G = vaccination with ChAdOx1 GFP; E = exam; C = challenge and exam; N = exam and necropsy (B) Two-fold serial-diluted serum samples were tested for MERS-CoV S-specific antibodies using ELISA. (C) ELISA-positive two-fold serial-diluted serum samples were tested for neutralizing antibodies against MERS-CoV in VeroE6 cells. Line = geometric mean, dotted line = limit of detection. Statistical significance between -28 DPI and -14 DPI in the prime-boost group was determined via one-tailed paired Student’s t-test. ** = p-value

    Journal: bioRxiv

    Article Title: A single dose of ChAdOx1 MERS provides broad protective immunity against a variety of MERS-CoV strains

    doi: 10.1101/2020.04.13.036293

    Figure Lengend Snippet: Vaccination of rhesus macaques with ChAdOx1 MERS elicits a humoral immune response. Serum samples were collected from non-human primates at times of vaccination (−56 DPI and -28 DPI), 14 days later and at challenge. (A) Overview of experimental timeline. V-M = vaccination with ChAdOx1 MERS; V-G = vaccination with ChAdOx1 GFP; E = exam; C = challenge and exam; N = exam and necropsy (B) Two-fold serial-diluted serum samples were tested for MERS-CoV S-specific antibodies using ELISA. (C) ELISA-positive two-fold serial-diluted serum samples were tested for neutralizing antibodies against MERS-CoV in VeroE6 cells. Line = geometric mean, dotted line = limit of detection. Statistical significance between -28 DPI and -14 DPI in the prime-boost group was determined via one-tailed paired Student’s t-test. ** = p-value

    Article Snippet: Specific anti-CoV immunoreactivity was detected using MERS-CoV nucleocapsid protein rabbit antibody (Sino Biological Inc.) at a 1:4000.

    Techniques: Enzyme-linked Immunosorbent Assay, One-tailed Test

    Representative HE findings in respiratory epithelium of the nasal turbinates of MERS-CoV-infected alpacas and immunohistochemical reactions of the commercially available positive controls for comparison. (A) Multifocal infiltration of lamina propria and submucosa by moderate numbers of lymphocytes, macrophages, and single neutrophilic granulocytes (asterisk). (B) Exocytosis of single neutrophilic granulocytes (grey arrow). (C, D) Abundant viral antigen was detected multifocally in the cytoplasm and along the apical membranous region of epithelial cells using a monoclonal mouse (C, Sino Biological Inc.) and polyclonal rabbit anti-MERS-CoV nucleocapsid antibody (D, Sino Biological Inc.) with citrate pretreatment, in a dilution of 1:70 and 1:2,000, respectively. Both antibodies exhibited a similar staining intensity. (A, B) HE staining; 400x, (C, D) Avidin-biotin-peroxidase complex method with 3,3′-diaminobenzidine as chromogen; 400x.

    Journal: Veterinary Immunology and Immunopathology

    Article Title: Detection of MERS-CoV antigen on formalin-fixed paraffin-embedded nasal tissue of alpacas by immunohistochemistry using human monoclonal antibodies directed against different epitopes of the spike protein

    doi: 10.1016/j.vetimm.2019.109939

    Figure Lengend Snippet: Representative HE findings in respiratory epithelium of the nasal turbinates of MERS-CoV-infected alpacas and immunohistochemical reactions of the commercially available positive controls for comparison. (A) Multifocal infiltration of lamina propria and submucosa by moderate numbers of lymphocytes, macrophages, and single neutrophilic granulocytes (asterisk). (B) Exocytosis of single neutrophilic granulocytes (grey arrow). (C, D) Abundant viral antigen was detected multifocally in the cytoplasm and along the apical membranous region of epithelial cells using a monoclonal mouse (C, Sino Biological Inc.) and polyclonal rabbit anti-MERS-CoV nucleocapsid antibody (D, Sino Biological Inc.) with citrate pretreatment, in a dilution of 1:70 and 1:2,000, respectively. Both antibodies exhibited a similar staining intensity. (A, B) HE staining; 400x, (C, D) Avidin-biotin-peroxidase complex method with 3,3′-diaminobenzidine as chromogen; 400x.

    Article Snippet: For positive control and elucidation of virus distribution, additional sections were stained with two different commercially available, previously published monoclonal mouse and polyclonal rabbit anti-MERS-CoV nucleocapsid antibodies (Sino Biological Inc.; ; ), respectively.

    Techniques: Infection, Immunohistochemistry, Staining, Avidin-Biotin Assay

    Representative positive immunohistochemical reactions for human monoclonal antibodies. (A) Antibody 1.2g5 with citrate pretreatment exhibited a strong multifocal MERS-CoV antigen specific signal in the cytoplasm of ciliated respiratory epithelial cells (arrow) and segmentally along the apical membranous region (arrow heads). (B) The same protocol without pretreatment revealed a much fainter but similarly distributed staining of cytoplasm (arrow) and ciliary base (arrow heads). (C) Antibody 1.10f3 with citrate pretreatment displayed a strong diffuse background staining and only few positively stained cilia (arrowhead). Single intraepithelial plasma cells displayed a strong false positive intracytoplasmic staining (white arrow). (D) Antibody 1.6c7 with citrate pretreatment exhibited also a strong diffuse background staining and only few positive staining cilia (arrow heads). (A–D) Avidin-biotin-peroxidase complex method with 3,3′-diaminobenzidine as chromogen; 400x.

    Journal: Veterinary Immunology and Immunopathology

    Article Title: Detection of MERS-CoV antigen on formalin-fixed paraffin-embedded nasal tissue of alpacas by immunohistochemistry using human monoclonal antibodies directed against different epitopes of the spike protein

    doi: 10.1016/j.vetimm.2019.109939

    Figure Lengend Snippet: Representative positive immunohistochemical reactions for human monoclonal antibodies. (A) Antibody 1.2g5 with citrate pretreatment exhibited a strong multifocal MERS-CoV antigen specific signal in the cytoplasm of ciliated respiratory epithelial cells (arrow) and segmentally along the apical membranous region (arrow heads). (B) The same protocol without pretreatment revealed a much fainter but similarly distributed staining of cytoplasm (arrow) and ciliary base (arrow heads). (C) Antibody 1.10f3 with citrate pretreatment displayed a strong diffuse background staining and only few positively stained cilia (arrowhead). Single intraepithelial plasma cells displayed a strong false positive intracytoplasmic staining (white arrow). (D) Antibody 1.6c7 with citrate pretreatment exhibited also a strong diffuse background staining and only few positive staining cilia (arrow heads). (A–D) Avidin-biotin-peroxidase complex method with 3,3′-diaminobenzidine as chromogen; 400x.

    Article Snippet: For positive control and elucidation of virus distribution, additional sections were stained with two different commercially available, previously published monoclonal mouse and polyclonal rabbit anti-MERS-CoV nucleocapsid antibodies (Sino Biological Inc.; ; ), respectively.

    Techniques: Immunohistochemistry, Staining, Avidin-Biotin Assay

    Representative negative immunohistochemical reactions for human monoclonal antibodies. (A–E) Specific staining for MERS-CoV antigen was absent for the antibodies 1.6f9 (A), 4.6e10 (B), 7.7g6 (C), 1.8e5 (D), and 3.5g6 (E), respectively. Reactions were accompanied by mild to severe non-specific, intracytoplasmic background staining. (A–E) Avidin-biotin-peroxidase complex method with 3,3′-diaminobenzidine as chromogen; 400x.

    Journal: Veterinary Immunology and Immunopathology

    Article Title: Detection of MERS-CoV antigen on formalin-fixed paraffin-embedded nasal tissue of alpacas by immunohistochemistry using human monoclonal antibodies directed against different epitopes of the spike protein

    doi: 10.1016/j.vetimm.2019.109939

    Figure Lengend Snippet: Representative negative immunohistochemical reactions for human monoclonal antibodies. (A–E) Specific staining for MERS-CoV antigen was absent for the antibodies 1.6f9 (A), 4.6e10 (B), 7.7g6 (C), 1.8e5 (D), and 3.5g6 (E), respectively. Reactions were accompanied by mild to severe non-specific, intracytoplasmic background staining. (A–E) Avidin-biotin-peroxidase complex method with 3,3′-diaminobenzidine as chromogen; 400x.

    Article Snippet: For positive control and elucidation of virus distribution, additional sections were stained with two different commercially available, previously published monoclonal mouse and polyclonal rabbit anti-MERS-CoV nucleocapsid antibodies (Sino Biological Inc.; ; ), respectively.

    Techniques: Immunohistochemistry, Staining, Avidin-Biotin Assay

    IHC detection of virus antigen expression in mouse tissue after challenge with MERS-CoV. Lung (A–C) and trachea (D–F) sections were assessed using rabbit polyclonal antibody to MERS-CoV nucleoprotein (NP) 3 days after the MERS-CoV challenge. The dark purple spot marked the inflammatory cell infiltration, and the brown particle marked the antigen of MERS-CoV. The MERS-CoV was located mainly in the trachea. Additionally, the lung tissue showed MERS-CoV expression in all immunized groups.

    Journal: Vaccine

    Article Title: The recombinant N-terminal domain of spike proteins is a potential vaccine against Middle East respiratory syndrome coronavirus (MERS-CoV) infection

    doi: 10.1016/j.vaccine.2016.11.064

    Figure Lengend Snippet: IHC detection of virus antigen expression in mouse tissue after challenge with MERS-CoV. Lung (A–C) and trachea (D–F) sections were assessed using rabbit polyclonal antibody to MERS-CoV nucleoprotein (NP) 3 days after the MERS-CoV challenge. The dark purple spot marked the inflammatory cell infiltration, and the brown particle marked the antigen of MERS-CoV. The MERS-CoV was located mainly in the trachea. Additionally, the lung tissue showed MERS-CoV expression in all immunized groups.

    Article Snippet: Rabbit-serum-derived polyclonal antibody against nucleoprotein (cat: 100213-RP02; Sino Biological Inc., Beijing, CHN) was incubated with the sections at 1:1000 dilution; goat anti-rabbit (cat: pv-9001; ZSGB-Bio, Beijing, CHN) secondary antibody was used at 1:2000, and sections were evaluated using light microscopy.

    Techniques: Immunohistochemistry, Expressing

    rNTD or rRBD vaccination reduced respiratory tract pathology in mice after MERS-CoV challenge. Representative results of hematoxylin-eosin (HE) staining in the lung (A – C) and trachea (D – F) of mock-treated or immunized mice. Severe lesions including the loss of pulmonary alveolus (represented by the white vacuole) and diffuse inflammatory cell infiltration (represented by the dark purple point) are shown (figure A). In contrast, milder lesions were observed among mice immunized with rRBD (figure B) or NTD (figure C), as the pulmonary alveolus was highly visible with less inflammatory cell infiltration. Inflammatory cell infiltration and impaired epithelium of the tunica mucosa bronchiorum were seen in the mock group (D). rRBD (E) or rNTD (F) alleviated the pathologic damage in the trachea of immunized mice. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

    Journal: Vaccine

    Article Title: The recombinant N-terminal domain of spike proteins is a potential vaccine against Middle East respiratory syndrome coronavirus (MERS-CoV) infection

    doi: 10.1016/j.vaccine.2016.11.064

    Figure Lengend Snippet: rNTD or rRBD vaccination reduced respiratory tract pathology in mice after MERS-CoV challenge. Representative results of hematoxylin-eosin (HE) staining in the lung (A – C) and trachea (D – F) of mock-treated or immunized mice. Severe lesions including the loss of pulmonary alveolus (represented by the white vacuole) and diffuse inflammatory cell infiltration (represented by the dark purple point) are shown (figure A). In contrast, milder lesions were observed among mice immunized with rRBD (figure B) or NTD (figure C), as the pulmonary alveolus was highly visible with less inflammatory cell infiltration. Inflammatory cell infiltration and impaired epithelium of the tunica mucosa bronchiorum were seen in the mock group (D). rRBD (E) or rNTD (F) alleviated the pathologic damage in the trachea of immunized mice. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

    Article Snippet: Rabbit-serum-derived polyclonal antibody against nucleoprotein (cat: 100213-RP02; Sino Biological Inc., Beijing, CHN) was incubated with the sections at 1:1000 dilution; goat anti-rabbit (cat: pv-9001; ZSGB-Bio, Beijing, CHN) secondary antibody was used at 1:2000, and sections were evaluated using light microscopy.

    Techniques: Mouse Assay, Staining

    Description of the N-terminal domain (NTD) immunogen and vaccination schedule. (A) The location of the NTD protein on the Middle East respiratory syndrome coronavirus MERS-CoV spike (S) protein. The recombinant (r)NTD protein consists of 336 amino acid (aa) residues (18–353) of S protein. A gp67 signal peptide (SP) was added to the N terminus for expression of the rNTD protein. (B) Purified rNTD protein detected by SDS-PAGE (left) and Western blot (right). The purified rNTD protein was separated by a 10% SDS-PAGE and stained with 0.25% Coomassie brilliant blue. Anti-NTD polyclonal antibody and infrared ray-labeled secondary antibody were used for the Western blot assay. Lane 1: protein molecular weight marker; lane 2: purified rNTD protein. (C). Vaccination schedule and detection. Mice received three vaccinations consisting of 5 or 10 μg of rNTD protein combined with adjuvants at 4-week intervals. Sera were collected at the indicated times to analyze the humoral immune response. Six mice from each group were sacrificed 2 weeks after the last immunization. The spleens were harvested for enzyme-linked immunospot (ELISpot), intracellular cytokine staining (ICS), and cytometric bead array (CBA) assays. In parallel experiments, the remaining mice were challenged with MERS-CoV to detect the protective effect elicited by the rNTD protein. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

    Journal: Vaccine

    Article Title: The recombinant N-terminal domain of spike proteins is a potential vaccine against Middle East respiratory syndrome coronavirus (MERS-CoV) infection

    doi: 10.1016/j.vaccine.2016.11.064

    Figure Lengend Snippet: Description of the N-terminal domain (NTD) immunogen and vaccination schedule. (A) The location of the NTD protein on the Middle East respiratory syndrome coronavirus MERS-CoV spike (S) protein. The recombinant (r)NTD protein consists of 336 amino acid (aa) residues (18–353) of S protein. A gp67 signal peptide (SP) was added to the N terminus for expression of the rNTD protein. (B) Purified rNTD protein detected by SDS-PAGE (left) and Western blot (right). The purified rNTD protein was separated by a 10% SDS-PAGE and stained with 0.25% Coomassie brilliant blue. Anti-NTD polyclonal antibody and infrared ray-labeled secondary antibody were used for the Western blot assay. Lane 1: protein molecular weight marker; lane 2: purified rNTD protein. (C). Vaccination schedule and detection. Mice received three vaccinations consisting of 5 or 10 μg of rNTD protein combined with adjuvants at 4-week intervals. Sera were collected at the indicated times to analyze the humoral immune response. Six mice from each group were sacrificed 2 weeks after the last immunization. The spleens were harvested for enzyme-linked immunospot (ELISpot), intracellular cytokine staining (ICS), and cytometric bead array (CBA) assays. In parallel experiments, the remaining mice were challenged with MERS-CoV to detect the protective effect elicited by the rNTD protein. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

    Article Snippet: Rabbit-serum-derived polyclonal antibody against nucleoprotein (cat: 100213-RP02; Sino Biological Inc., Beijing, CHN) was incubated with the sections at 1:1000 dilution; goat anti-rabbit (cat: pv-9001; ZSGB-Bio, Beijing, CHN) secondary antibody was used at 1:2000, and sections were evaluated using light microscopy.

    Techniques: Recombinant, Expressing, Purification, SDS Page, Western Blot, Staining, Labeling, Molecular Weight, Marker, Mouse Assay, Enzyme-linked Immunospot, Crocin Bleaching Assay

    Kinetics of serum and egg yolk anti-MERS COV-S IgY antibodies response of chickens after immunization with MERS COV-S recombinant protein compared with the adjuvant-immunized chicken (adjuvant control). Each week is represented by a pool of egg yolks of individual chicken in each group (S1-immunized and adjuvant-immunized).

    Journal: Vaccines

    Article Title: Anti-S1 MERS-COV IgY Specific Antibodies Decreases Lung Inflammation and Viral Antigen Positive Cells in the Human Transgenic Mouse Model

    doi: 10.3390/vaccines8040634

    Figure Lengend Snippet: Kinetics of serum and egg yolk anti-MERS COV-S IgY antibodies response of chickens after immunization with MERS COV-S recombinant protein compared with the adjuvant-immunized chicken (adjuvant control). Each week is represented by a pool of egg yolks of individual chicken in each group (S1-immunized and adjuvant-immunized).

    Article Snippet: MERS-CoV antigens were detected utilizing a polymer-based detection system (Nichirei-Histofine Simple Stain Mouse MAX PO(R); Nichirei) with a rabbit anti-MERS-CoV nucleocapsid antibody (40068-RP01; Sino Biological Inc., Beijing, China).

    Techniques: Recombinant

    Recognition by anti-S1 IgY antibodies of viral antigen expressed in MERS-CoV-infected Vero E6 cells, using indirect immunofluorescence assay. ( A ) Vero E6 cells inoculated with MERS-CoV and stained with anti-S1 IgY antibodies and FITC-conjugated anti-chicken antibodies; and ( B ) control adjuvant IgY (Bright-field).

    Journal: Vaccines

    Article Title: Anti-S1 MERS-COV IgY Specific Antibodies Decreases Lung Inflammation and Viral Antigen Positive Cells in the Human Transgenic Mouse Model

    doi: 10.3390/vaccines8040634

    Figure Lengend Snippet: Recognition by anti-S1 IgY antibodies of viral antigen expressed in MERS-CoV-infected Vero E6 cells, using indirect immunofluorescence assay. ( A ) Vero E6 cells inoculated with MERS-CoV and stained with anti-S1 IgY antibodies and FITC-conjugated anti-chicken antibodies; and ( B ) control adjuvant IgY (Bright-field).

    Article Snippet: MERS-CoV antigens were detected utilizing a polymer-based detection system (Nichirei-Histofine Simple Stain Mouse MAX PO(R); Nichirei) with a rabbit anti-MERS-CoV nucleocapsid antibody (40068-RP01; Sino Biological Inc., Beijing, China).

    Techniques: Infection, Immunofluorescence, Staining

    Dot blotting analysis. Purified anti-S1 IgY antibodies showed reactivity with different concentrations of the spike protein (S), S1, and receptor binding domain (RBD), but had no reactivity with nucleocapsid (NP) protein of MERS CoV.

    Journal: Vaccines

    Article Title: Anti-S1 MERS-COV IgY Specific Antibodies Decreases Lung Inflammation and Viral Antigen Positive Cells in the Human Transgenic Mouse Model

    doi: 10.3390/vaccines8040634

    Figure Lengend Snippet: Dot blotting analysis. Purified anti-S1 IgY antibodies showed reactivity with different concentrations of the spike protein (S), S1, and receptor binding domain (RBD), but had no reactivity with nucleocapsid (NP) protein of MERS CoV.

    Article Snippet: MERS-CoV antigens were detected utilizing a polymer-based detection system (Nichirei-Histofine Simple Stain Mouse MAX PO(R); Nichirei) with a rabbit anti-MERS-CoV nucleocapsid antibody (40068-RP01; Sino Biological Inc., Beijing, China).

    Techniques: Purification, Binding Assay

    Examples of different concentrations of anti-S1 IgY antibodies tested against MERS-CoV on Vero-E6 cells examined by CPE. The IC100 neutralization of the antibody were determined as the reciprocal of the highest dilution at which no CPE was observed.

    Journal: Vaccines

    Article Title: Anti-S1 MERS-COV IgY Specific Antibodies Decreases Lung Inflammation and Viral Antigen Positive Cells in the Human Transgenic Mouse Model

    doi: 10.3390/vaccines8040634

    Figure Lengend Snippet: Examples of different concentrations of anti-S1 IgY antibodies tested against MERS-CoV on Vero-E6 cells examined by CPE. The IC100 neutralization of the antibody were determined as the reciprocal of the highest dilution at which no CPE was observed.

    Article Snippet: MERS-CoV antigens were detected utilizing a polymer-based detection system (Nichirei-Histofine Simple Stain Mouse MAX PO(R); Nichirei) with a rabbit anti-MERS-CoV nucleocapsid antibody (40068-RP01; Sino Biological Inc., Beijing, China).

    Techniques: Neutralization

    ( A ) Viral titer in the lungs of MERS-CoV mice treated with anti-SI IgY antibodies and control IgY (adjuvant). ( B ) Body weight changes after MERS-CoV infection between anti-SI IgY antibodies and IgY of adjuvant control group. ( C – F ) Histopathology of the lungs from human dipeptidyl peptidase 4 (hDPP4)-transgenic mice on day 8 after inoculation with MERS-CoV. Representative histopathological findings of mice with the highest cellular infiltration in alveoli by H E staining ( C ) Massive mononuclear cell infiltrations including macrophages and lymphocytes with regenerated type II pneumocytes were seen in adjuvant control group (right column), but less in the anti-S1 IgY treated group (left column). Scale bars: 200 μm (upper row) and 20 μm (lower row). Al, alveoli; Br, bronchi; V, vessel. Detection of viral antigen in lung tissues of mice by immunohistochemistry ( D ) A few antigen positive cells were seen in the lungs of anti-S1 IgY treated group compared to adjuvant control group. Quantification of inflammation areas ( E ) The area of pulmonary lesion was determined based on the mean percentage of affected areas in each section of the collected lobes form each animal ( n = 8 or 6). Circles indicate averages from three observation lobes in each mouse. p = 0.1709 by Mann-Whitney test. Numbers of viral antigen positive cells in the alveoli ( F ) Data were obtained from 8 or 6 mice. Circles indicate averages of 5 observation fields in each mouse. * p = 0.0196 by Mann-Whitney test.

    Journal: Vaccines

    Article Title: Anti-S1 MERS-COV IgY Specific Antibodies Decreases Lung Inflammation and Viral Antigen Positive Cells in the Human Transgenic Mouse Model

    doi: 10.3390/vaccines8040634

    Figure Lengend Snippet: ( A ) Viral titer in the lungs of MERS-CoV mice treated with anti-SI IgY antibodies and control IgY (adjuvant). ( B ) Body weight changes after MERS-CoV infection between anti-SI IgY antibodies and IgY of adjuvant control group. ( C – F ) Histopathology of the lungs from human dipeptidyl peptidase 4 (hDPP4)-transgenic mice on day 8 after inoculation with MERS-CoV. Representative histopathological findings of mice with the highest cellular infiltration in alveoli by H E staining ( C ) Massive mononuclear cell infiltrations including macrophages and lymphocytes with regenerated type II pneumocytes were seen in adjuvant control group (right column), but less in the anti-S1 IgY treated group (left column). Scale bars: 200 μm (upper row) and 20 μm (lower row). Al, alveoli; Br, bronchi; V, vessel. Detection of viral antigen in lung tissues of mice by immunohistochemistry ( D ) A few antigen positive cells were seen in the lungs of anti-S1 IgY treated group compared to adjuvant control group. Quantification of inflammation areas ( E ) The area of pulmonary lesion was determined based on the mean percentage of affected areas in each section of the collected lobes form each animal ( n = 8 or 6). Circles indicate averages from three observation lobes in each mouse. p = 0.1709 by Mann-Whitney test. Numbers of viral antigen positive cells in the alveoli ( F ) Data were obtained from 8 or 6 mice. Circles indicate averages of 5 observation fields in each mouse. * p = 0.0196 by Mann-Whitney test.

    Article Snippet: MERS-CoV antigens were detected utilizing a polymer-based detection system (Nichirei-Histofine Simple Stain Mouse MAX PO(R); Nichirei) with a rabbit anti-MERS-CoV nucleocapsid antibody (40068-RP01; Sino Biological Inc., Beijing, China).

    Techniques: Mouse Assay, Infection, Histopathology, Transgenic Assay, Staining, Immunohistochemistry, MANN-WHITNEY

    Western blot analysis of anti-MERS-COV rS1 IgY antibodies. (Left) The S1 protein of MERS-COV was subjected to SDS-PAGE under reducing conditions; (Right) Western blot analysis of the anti-S1 IgY antibody response. SDS gels were electrically transferred onto nitrocellulose membranes and probed with IgY from immunized and nonimmunized hens (marker: molecular maker; lane A: S1-immunized IgY; lane B: adjuvant-immunized IgY). The strips were processed separately and pasted beside each other for documentation.

    Journal: Vaccines

    Article Title: Anti-S1 MERS-COV IgY Specific Antibodies Decreases Lung Inflammation and Viral Antigen Positive Cells in the Human Transgenic Mouse Model

    doi: 10.3390/vaccines8040634

    Figure Lengend Snippet: Western blot analysis of anti-MERS-COV rS1 IgY antibodies. (Left) The S1 protein of MERS-COV was subjected to SDS-PAGE under reducing conditions; (Right) Western blot analysis of the anti-S1 IgY antibody response. SDS gels were electrically transferred onto nitrocellulose membranes and probed with IgY from immunized and nonimmunized hens (marker: molecular maker; lane A: S1-immunized IgY; lane B: adjuvant-immunized IgY). The strips were processed separately and pasted beside each other for documentation.

    Article Snippet: MERS-CoV antigens were detected utilizing a polymer-based detection system (Nichirei-Histofine Simple Stain Mouse MAX PO(R); Nichirei) with a rabbit anti-MERS-CoV nucleocapsid antibody (40068-RP01; Sino Biological Inc., Beijing, China).

    Techniques: Western Blot, SDS Page, Marker

    Evaluation of the neutralizing potential of anti-S1 IgY antibodies, using plaque reduction neutralization test. ( A ) MERS-CoV (MOI 0.01) was incubated with different concentrations of anti-S1 IgY antibodies and added to Vero E6 cells. After virus adsorption, agar medium was added to the Vero E6 cells, and the plaques that formed were stained with crystal violet, each IgY concentration was tested in triplicate. ( B ) Percent inhibition of anti-S1 IgY antibodies with different concentrations. The best fit equation is:

    Journal: Vaccines

    Article Title: Anti-S1 MERS-COV IgY Specific Antibodies Decreases Lung Inflammation and Viral Antigen Positive Cells in the Human Transgenic Mouse Model

    doi: 10.3390/vaccines8040634

    Figure Lengend Snippet: Evaluation of the neutralizing potential of anti-S1 IgY antibodies, using plaque reduction neutralization test. ( A ) MERS-CoV (MOI 0.01) was incubated with different concentrations of anti-S1 IgY antibodies and added to Vero E6 cells. After virus adsorption, agar medium was added to the Vero E6 cells, and the plaques that formed were stained with crystal violet, each IgY concentration was tested in triplicate. ( B ) Percent inhibition of anti-S1 IgY antibodies with different concentrations. The best fit equation is:

    Article Snippet: MERS-CoV antigens were detected utilizing a polymer-based detection system (Nichirei-Histofine Simple Stain Mouse MAX PO(R); Nichirei) with a rabbit anti-MERS-CoV nucleocapsid antibody (40068-RP01; Sino Biological Inc., Beijing, China).

    Techniques: Plaque Reduction Neutralization Test, Incubation, Adsorption, Staining, Concentration Assay, Inhibition