mers cov nucleocapsid protein rabbit antibody  (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 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
    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 rabbit antibody/product/Sino Biological
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mers cov nucleocapsid protein rabbit antibody - by Bioz Stars, 2021-06
    95/100 stars

    Images

    1) Product Images from "A single dose of ChAdOx1 MERS provides broad protective immunity against a variety of MERS-CoV strains"

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

    Journal: bioRxiv

    doi: 10.1101/2020.04.13.036293

    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
    Figure Legend 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

    Techniques Used: 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.
    Figure Legend 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.

    Techniques Used: 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.
    Figure Legend 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.

    Techniques Used:

    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
    Figure Legend 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

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

    2) Product Images from "Surface-aerosol stability and pathogenicity of diverse MERS-CoV strains from 2012 - 2018"

    Article Title: Surface-aerosol stability and pathogenicity of diverse MERS-CoV strains from 2012 - 2018

    Journal: bioRxiv

    doi: 10.1101/2021.02.11.429193

    In vivo replication of different MERS-CoV strains. hDPP4 mice were inoculated I.N. with 10 3 TCID 50 MERS-CoV. Four mice were euthanized on D3, and the remaining 6 mice were monitored for survival. A.) Relative weight loss of hDPP4 mice. B.) Survival of hDPP4 mice. C.) Oropharyngeal shedding of MERS-CoV as measured via UpE qRT-PCR. D.) Area under the curve of oropharyngeal MERS-CoV shedding per virus strain. E.) Viral load in lung tissue obtained from mice euthanized at D3. F.) Viral mRNA load in lung tissue obtained from mice euthanized at D3. G.) Lung tissue were stained for MERS-CoV antigen and % of positive pixels was quantified. Statistical significance was compared using an unpaired two-tailed Student’s t-test. P-value = *
    Figure Legend Snippet: In vivo replication of different MERS-CoV strains. hDPP4 mice were inoculated I.N. with 10 3 TCID 50 MERS-CoV. Four mice were euthanized on D3, and the remaining 6 mice were monitored for survival. A.) Relative weight loss of hDPP4 mice. B.) Survival of hDPP4 mice. C.) Oropharyngeal shedding of MERS-CoV as measured via UpE qRT-PCR. D.) Area under the curve of oropharyngeal MERS-CoV shedding per virus strain. E.) Viral load in lung tissue obtained from mice euthanized at D3. F.) Viral mRNA load in lung tissue obtained from mice euthanized at D3. G.) Lung tissue were stained for MERS-CoV antigen and % of positive pixels was quantified. Statistical significance was compared using an unpaired two-tailed Student’s t-test. P-value = *

    Techniques Used: In Vivo, Mouse Assay, Quantitative RT-PCR, Staining, Two Tailed Test

    Phylogenetic tree of MERS-CoV strains. Maximum likelihood tree of 446 full MERS CoV genomes showing distribution of isolates used in this study. Human-derived MERS-CoV isolates used in this study are highlighted in red, camel-derived MERS-CoV isolates are highlighted in blue. Phylogenetic tree reconstructed with PhyML and rooted at the midpoint.
    Figure Legend Snippet: Phylogenetic tree of MERS-CoV strains. Maximum likelihood tree of 446 full MERS CoV genomes showing distribution of isolates used in this study. Human-derived MERS-CoV isolates used in this study are highlighted in red, camel-derived MERS-CoV isolates are highlighted in blue. Phylogenetic tree reconstructed with PhyML and rooted at the midpoint.

    Techniques Used: Derivative Assay

    Stability of MERS-CoV strains on surfaces and in aerosols compared to SARS-CoV-2. A.) 50 µl of MERS-CoV or SARS-CoV-2 was spread on surface, either polypropylene, stainless steel, copper or silver. 1 mL of DMEM was added at T=0, 1, 24, 48 or 72 hours and titrated. B.) MERS-CoV or SARS-CoV-2 containing aerosols were sprayed into the Goldberg drum, and samples were taken at T=0, 30, 60, 120 and 180 minutes and titrated. Linear regression was calculated per virus and displayed in the graph as a line. A-B.) Statistically significant differences between EMC/12 and other strains were calculated using an unpaired Student’s two-tailed t-test corrected for multiple comparisons via Bonferroni. Dotted line = limit of detection; p-values = *
    Figure Legend Snippet: Stability of MERS-CoV strains on surfaces and in aerosols compared to SARS-CoV-2. A.) 50 µl of MERS-CoV or SARS-CoV-2 was spread on surface, either polypropylene, stainless steel, copper or silver. 1 mL of DMEM was added at T=0, 1, 24, 48 or 72 hours and titrated. B.) MERS-CoV or SARS-CoV-2 containing aerosols were sprayed into the Goldberg drum, and samples were taken at T=0, 30, 60, 120 and 180 minutes and titrated. Linear regression was calculated per virus and displayed in the graph as a line. A-B.) Statistically significant differences between EMC/12 and other strains were calculated using an unpaired Student’s two-tailed t-test corrected for multiple comparisons via Bonferroni. Dotted line = limit of detection; p-values = *

    Techniques Used: Two Tailed Test

    3) Product Images from "A single dose of ChAdOx1 MERS provides protective immunity in rhesus macaques"

    Article Title: A single dose of ChAdOx1 MERS provides protective immunity in rhesus macaques

    Journal: Science Advances

    doi: 10.1126/sciadv.aba8399

    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 the 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 (D), and mean with SD is shown (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, and mean with SD is shown. ( 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 (arrows) in lung tissue of animals vaccinated with ChAdOx1 MERS. Marked bronchointerstitial pneumonia with abundant pulmonary edema and fibrin (asterisks), type II pneumocyte hyperplasia (arrows), and increased numbers of alveolar macrophages (arrowheads) in lung tissue of control animals. Magnification, ×200. ( F ) Lung–to–body weight (BW) ratio was determined for all animals at necropsy. Mean with SD is shown. ( 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 to 10%; 2, 11 to 25%; 3, 26 to 50%; 4, 51 to 75%; and 5, 76 to 100%) and averaged per group. Mean with SD is shown. A, bronchointerstitial pneumonia; B, type II pneumocyte hyperplasia; C, hemorrhages, edema, and fibrin deposits. Statistical significance between groups was determined via two-tailed unpaired Student’s t test. * P
    Figure Legend 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 the 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 (D), and mean with SD is shown (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, and mean with SD is shown. ( 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 (arrows) in lung tissue of animals vaccinated with ChAdOx1 MERS. Marked bronchointerstitial pneumonia with abundant pulmonary edema and fibrin (asterisks), type II pneumocyte hyperplasia (arrows), and increased numbers of alveolar macrophages (arrowheads) in lung tissue of control animals. Magnification, ×200. ( F ) Lung–to–body weight (BW) ratio was determined for all animals at necropsy. Mean with SD is shown. ( 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 to 10%; 2, 11 to 25%; 3, 26 to 50%; 4, 51 to 75%; and 5, 76 to 100%) and averaged per group. Mean with SD is shown. A, bronchointerstitial pneumonia; B, type II pneumocyte hyperplasia; C, hemorrhages, edema, and fibrin deposits. Statistical significance between groups was determined via two-tailed unpaired Student’s t test. * P

    Techniques Used: Marker, Staining, Two Tailed Test

    Vaccination of rhesus macaques with ChAdOx1 MERS elicits a humoral immune response. Serum samples were collected from NHPs at times of vaccination (−56 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 ) Twofold serial diluted serum samples were tested for MERS-CoV S–specific antibodies using enzyme-linked immunosorbent assay (ELISA). ( C ) Twofold serial diluted serum samples were tested for neutralizing antibodies against MERS-CoV in VeroE6 cells. Line, geometric mean; dotted line, limit of detection (LoD). Statistical significance between −28 and −14 DPI in the prime-boost group was determined via one-tailed paired Student’s t test. Statistical significance between prime-boost and prime-only groups on 0 DPI was determined via two-tailed unpaired Student’s t test. ** P
    Figure Legend Snippet: Vaccination of rhesus macaques with ChAdOx1 MERS elicits a humoral immune response. Serum samples were collected from NHPs at times of vaccination (−56 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 ) Twofold serial diluted serum samples were tested for MERS-CoV S–specific antibodies using enzyme-linked immunosorbent assay (ELISA). ( C ) Twofold serial diluted serum samples were tested for neutralizing antibodies against MERS-CoV in VeroE6 cells. Line, geometric mean; dotted line, limit of detection (LoD). Statistical significance between −28 and −14 DPI in the prime-boost group was determined via one-tailed paired Student’s t test. Statistical significance between prime-boost and prime-only groups on 0 DPI was determined via two-tailed unpaired Student’s t test. ** P

    Techniques Used: Enzyme-linked Immunosorbent Assay, One-tailed Test, 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 at 3 DPI from hDPP4 mice challenged with MERS-CoV. Mean titer with SD is shown.
    Figure Legend 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 at 3 DPI from hDPP4 mice challenged with MERS-CoV. Mean titer with SD is shown.

    Techniques Used: Mouse Assay

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    Article Snippet: The assays were validated using pulmonary tissues obtained at autopsies from patients who died of COVID-19; the validation indicated equally high specificity of SARS-CoV-2 ISH and ICH with the highest sensitivity of the anti-nucleocapsid IHC. .. Sixty-four placentas from infected women were studied with at least one modality, including 4 cases with ISH, 21 with anti-spike and 39 with anti-nucleocapsid IHC, and 11 placentas were studied with combinations of the above techniques, including 2 with anti-nucleocapsid antibody and ISH, 5 with anti-spike antibody and ISH, 3 with anti-spike and anti-nucleocapsid antibodies, and 1 case was studied with all 3 techniques. .. The assays were carried out using Bond-III Fully Automated IHC and ISH Stainer (Leica Biosystems, Vista, CA) according to the manufacturers’ recommendations.

    In Situ Hybridization:

    Article Title: Trophoblast Damage with Acute and Chronic Intervillositis: Disruption of Placental Barrier by SARS-CoV-2
    Article Snippet: The assays were validated using pulmonary tissues obtained at autopsies from patients who died of COVID-19; the validation indicated equally high specificity of SARS-CoV-2 ISH and ICH with the highest sensitivity of the anti-nucleocapsid IHC. .. Sixty-four placentas from infected women were studied with at least one modality, including 4 cases with ISH, 21 with anti-spike and 39 with anti-nucleocapsid IHC, and 11 placentas were studied with combinations of the above techniques, including 2 with anti-nucleocapsid antibody and ISH, 5 with anti-spike antibody and ISH, 3 with anti-spike and anti-nucleocapsid antibodies, and 1 case was studied with all 3 techniques. .. The assays were carried out using Bond-III Fully Automated IHC and ISH Stainer (Leica Biosystems, Vista, CA) according to the manufacturers’ recommendations.

    Immunohistochemistry:

    Article Title: Trophoblast Damage with Acute and Chronic Intervillositis: Disruption of Placental Barrier by SARS-CoV-2
    Article Snippet: The assays were validated using pulmonary tissues obtained at autopsies from patients who died of COVID-19; the validation indicated equally high specificity of SARS-CoV-2 ISH and ICH with the highest sensitivity of the anti-nucleocapsid IHC. .. Sixty-four placentas from infected women were studied with at least one modality, including 4 cases with ISH, 21 with anti-spike and 39 with anti-nucleocapsid IHC, and 11 placentas were studied with combinations of the above techniques, including 2 with anti-nucleocapsid antibody and ISH, 5 with anti-spike antibody and ISH, 3 with anti-spike and anti-nucleocapsid antibodies, and 1 case was studied with all 3 techniques. .. The assays were carried out using Bond-III Fully Automated IHC and ISH Stainer (Leica Biosystems, Vista, CA) according to the manufacturers’ recommendations.

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    Incubation:

    Article Title: The recombinant N-terminal domain of spike proteins is a potential vaccine against Middle East respiratory syndrome coronavirus (MERS-CoV) infection
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    Article Title: Enhanced protection in mice induced by immunization with inactivated whole viruses compare to spike protein of middle east respiratory syndrome coronavirus
    Article Snippet: Tissues were fixed using 10% neutral buffered formalin, embedded in paraffin, sectioned sequentially to a 4 μm thickness, and stained with HE prior to examination by light microscopy. .. For immunohistochemistry (IHC), the sections were then incubated with a rabbit-serum-derived polyclonal antibody against nucleoprotein (Sino Biological, Inc., Cat. No. 100213-RP02) or mouse-serum-derived monoclonal antibody against S protein (produced in our laboratory) at a 1:1000 dilution. ..

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    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.

    Produced:

    Article Title: Enhanced protection in mice induced by immunization with inactivated whole viruses compare to spike protein of middle east respiratory syndrome coronavirus
    Article Snippet: Tissues were fixed using 10% neutral buffered formalin, embedded in paraffin, sectioned sequentially to a 4 μm thickness, and stained with HE prior to examination by light microscopy. .. For immunohistochemistry (IHC), the sections were then incubated with a rabbit-serum-derived polyclonal antibody against nucleoprotein (Sino Biological, Inc., Cat. No. 100213-RP02) or mouse-serum-derived monoclonal antibody against S protein (produced in our laboratory) at a 1:1000 dilution. ..

    other:

    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 polyclonal rabbit anti mers cov nucleocapsid antibodies
    Representative HE findings in respiratory epithelium of the nasal turbinates of <t>MERS-CoV-infected</t> 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 <t>polyclonal</t> 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.
    Polyclonal Rabbit Anti Mers Cov Nucleocapsid Antibodies, 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
    https://www.bioz.com/result/polyclonal rabbit anti mers cov nucleocapsid antibodies/product/Sino Biological
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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

    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

    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

    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