sars cov 2 1 2 spike protein 2b3e5  (Cell Signaling Technology Inc)


Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc sars cov 2 1 2 spike protein 2b3e5
    Specificities and sources of primary and secondary antibodies.
    Sars Cov 2 1 2 Spike Protein 2b3e5, supplied by Cell Signaling Technology Inc, 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/sars cov 2 1 2 spike protein 2b3e5/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 1 2 spike protein 2b3e5 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Development of a high-sensitivity and short-duration fluorescence in situ hybridization method for viral mRNA detection in HEK 293T cells"

    Article Title: Development of a high-sensitivity and short-duration fluorescence in situ hybridization method for viral mRNA detection in HEK 293T cells

    Journal: Frontiers in Cellular and Infection Microbiology

    doi: 10.3389/fcimb.2022.960938

    Specificities and sources of primary and secondary antibodies.
    Figure Legend Snippet: Specificities and sources of primary and secondary antibodies.

    Techniques Used:

    Detection of SARS-CoV-2 E mRNA and E/S protein in HEK 293T cells at 24 h after transfection with SARS-CoV-2 E and S plasmids. (A) SARS-CoV-2 E mRNA (with mixed probes containing BME001 and BME002), SARS-CoV-2 E, and SARS-CoV-2 S are shown in red, green, and blue, respectively. (B) Total mRNAs (detected by Oligo dT probes), SARS-CoV-2 E, and SARS-CoV-2 S are shown in red, green, and blue, respectively. (C) Negative controls by using secondary antibodies. Objective lens: 60×; Scale bar: 10 µm; Nu, nucleus.
    Figure Legend Snippet: Detection of SARS-CoV-2 E mRNA and E/S protein in HEK 293T cells at 24 h after transfection with SARS-CoV-2 E and S plasmids. (A) SARS-CoV-2 E mRNA (with mixed probes containing BME001 and BME002), SARS-CoV-2 E, and SARS-CoV-2 S are shown in red, green, and blue, respectively. (B) Total mRNAs (detected by Oligo dT probes), SARS-CoV-2 E, and SARS-CoV-2 S are shown in red, green, and blue, respectively. (C) Negative controls by using secondary antibodies. Objective lens: 60×; Scale bar: 10 µm; Nu, nucleus.

    Techniques Used: Transfection

    sars cov 2 1 2 spike protein 2b3e5  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc sars cov 2 1 2 spike protein 2b3e5
    Specificities and sources of primary and secondary antibodies.
    Sars Cov 2 1 2 Spike Protein 2b3e5, supplied by Cell Signaling Technology Inc, 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/sars cov 2 1 2 spike protein 2b3e5/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 1 2 spike protein 2b3e5 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Development of a high-sensitivity and short-duration fluorescence in situ hybridization method for viral mRNA detection in HEK 293T cells"

    Article Title: Development of a high-sensitivity and short-duration fluorescence in situ hybridization method for viral mRNA detection in HEK 293T cells

    Journal: Frontiers in Cellular and Infection Microbiology

    doi: 10.3389/fcimb.2022.960938

    Specificities and sources of primary and secondary antibodies.
    Figure Legend Snippet: Specificities and sources of primary and secondary antibodies.

    Techniques Used:

    Detection of SARS-CoV-2 E mRNA and E/S protein in HEK 293T cells at 24 h after transfection with SARS-CoV-2 E and S plasmids. (A) SARS-CoV-2 E mRNA (with mixed probes containing BME001 and BME002), SARS-CoV-2 E, and SARS-CoV-2 S are shown in red, green, and blue, respectively. (B) Total mRNAs (detected by Oligo dT probes), SARS-CoV-2 E, and SARS-CoV-2 S are shown in red, green, and blue, respectively. (C) Negative controls by using secondary antibodies. Objective lens: 60×; Scale bar: 10 µm; Nu, nucleus.
    Figure Legend Snippet: Detection of SARS-CoV-2 E mRNA and E/S protein in HEK 293T cells at 24 h after transfection with SARS-CoV-2 E and S plasmids. (A) SARS-CoV-2 E mRNA (with mixed probes containing BME001 and BME002), SARS-CoV-2 E, and SARS-CoV-2 S are shown in red, green, and blue, respectively. (B) Total mRNAs (detected by Oligo dT probes), SARS-CoV-2 E, and SARS-CoV-2 S are shown in red, green, and blue, respectively. (C) Negative controls by using secondary antibodies. Objective lens: 60×; Scale bar: 10 µm; Nu, nucleus.

    Techniques Used: Transfection

    sars cov 2 spike rbd  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc sars cov 2 spike rbd
    Prevalence of IgG specific for betacoronavirus RBDs in study cohort. Plasma was assayed by ELISA using recombinant, multimeric RBD as antigen. (a) <t>SARS-COV-2</t> spike 319-591 with p values from ANOVA with Tukey HSD for all pairwise comparisons shown beneath. (b) SARS-CoV spike 384-655 with p values shown as above, (c) MERS spike 306-577. Each point indicates an individual participant. Blue bars show median values. N = 7–30. (d) Comparison of absorbance values for MERS and SARS-CoV RBD binding IgG for all participants. Each point indicates an individual participant, n = 124. The orange circle indicates participants with detectable amounts of MERS RBD-binding IgG, showing correlation with SARS-CoV-binding IgG in this subset. IgG specific to recombinant, multimeric RBD from seasonal betacoronaviruses, (e) OC43 spike 315-675, (f) HKU1 spike 307-675 assayed by ELISA as above.
    Sars Cov 2 Spike Rbd, supplied by Cell Signaling Technology Inc, 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/sars cov 2 spike rbd/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 spike rbd - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Repertoires of SARS-CoV-2 epitopes targeted by antibodies vary according to severity of COVID-19"

    Article Title: Repertoires of SARS-CoV-2 epitopes targeted by antibodies vary according to severity of COVID-19

    Journal: Virulence

    doi: 10.1080/21505594.2022.2073025

    Prevalence of IgG specific for betacoronavirus RBDs in study cohort. Plasma was assayed by ELISA using recombinant, multimeric RBD as antigen. (a) SARS-COV-2 spike 319-591 with p values from ANOVA with Tukey HSD for all pairwise comparisons shown beneath. (b) SARS-CoV spike 384-655 with p values shown as above, (c) MERS spike 306-577. Each point indicates an individual participant. Blue bars show median values. N = 7–30. (d) Comparison of absorbance values for MERS and SARS-CoV RBD binding IgG for all participants. Each point indicates an individual participant, n = 124. The orange circle indicates participants with detectable amounts of MERS RBD-binding IgG, showing correlation with SARS-CoV-binding IgG in this subset. IgG specific to recombinant, multimeric RBD from seasonal betacoronaviruses, (e) OC43 spike 315-675, (f) HKU1 spike 307-675 assayed by ELISA as above.
    Figure Legend Snippet: Prevalence of IgG specific for betacoronavirus RBDs in study cohort. Plasma was assayed by ELISA using recombinant, multimeric RBD as antigen. (a) SARS-COV-2 spike 319-591 with p values from ANOVA with Tukey HSD for all pairwise comparisons shown beneath. (b) SARS-CoV spike 384-655 with p values shown as above, (c) MERS spike 306-577. Each point indicates an individual participant. Blue bars show median values. N = 7–30. (d) Comparison of absorbance values for MERS and SARS-CoV RBD binding IgG for all participants. Each point indicates an individual participant, n = 124. The orange circle indicates participants with detectable amounts of MERS RBD-binding IgG, showing correlation with SARS-CoV-binding IgG in this subset. IgG specific to recombinant, multimeric RBD from seasonal betacoronaviruses, (e) OC43 spike 315-675, (f) HKU1 spike 307-675 assayed by ELISA as above.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Recombinant, Binding Assay

    Prevalence of IgGs specific for SARS-CoV-2 peptides. (b) Individual peptides with significant variation between groups ( p < 0.05, ANOVA, n = 7–30). Bars denote groups with significant differences by pairwise comparison (Tukey HSD, p < 0.05).
    Figure Legend Snippet: Prevalence of IgGs specific for SARS-CoV-2 peptides. (b) Individual peptides with significant variation between groups ( p < 0.05, ANOVA, n = 7–30). Bars denote groups with significant differences by pairwise comparison (Tukey HSD, p < 0.05).

    Techniques Used:

    Comparison between peptide seropositivity frequencies in the primary dataset with an independent cohort. Peptides that showed >20 percentage point difference in frequency of seropositivity between participants in the Chelsea survey who had been hospitalized and those who reported exposure to SARS-CoV-2 are shown in green. The differences in frequency of seropositivity for IgG specific to those peptides between acutely hospitalized patients and people with no history of infection are shown in blue.
    Figure Legend Snippet: Comparison between peptide seropositivity frequencies in the primary dataset with an independent cohort. Peptides that showed >20 percentage point difference in frequency of seropositivity between participants in the Chelsea survey who had been hospitalized and those who reported exposure to SARS-CoV-2 are shown in green. The differences in frequency of seropositivity for IgG specific to those peptides between acutely hospitalized patients and people with no history of infection are shown in blue.

    Techniques Used: Infection

    sars cov 2 spike  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc sars cov 2 spike
    <t>SARS-CoV-2</t> caused lethal disease in K18-hACE2 mice irrespective of dose (10 4 vs. 10 6 PFU). K18-hACE2 mice (n = 50) were inoculated intranasally with either 1 × 10 4 or 1 × 10 6 plaque forming units (PFU), with n = 6 additional Sham/PBS negative controls. Body weight ( A ), clinical signs ( B ), body temperature ( C ), and survival ( D ) were monitored daily in Sham/PBS animals (black) and in infected animals (male, red; female, blue; up to 14 dpi). Mice meeting euthanasia criteria were counted dead the following day. Viral loads (viral RNA genome copy numbers/mg of tissue) or viral titers (infectious virus particles; PFU/mg of tissue) were quantified in the lung and brain ( E – G ). RNA copies were also examined in the serum (genome copies/mL) either directly on serum ( H ) or via a re-infectivity assay ( I ) using Vero E6 cells. The limit of detection is shown with a dashed line. Clinical data ( A – D ): 10 4 PFU; male (n = 9), female (n = 6); 10 6 PFU male (19), female (n = 16); Sham/PBS male (n = 3), female (n = 3). Molecular and virologic data ( E – I ). 10 6 RT-PCR: lung 2 dpi (n = 3), 4 dpi (n = 6), 7 dpi (n = 8); brain 2 dpi (n = 3), 4 dpi (n = 5), 7 dpi (n = 7). 10 6 PFU analysis: lung 2 dpi (n = 8), 4 dpi (n = 8), 7 dpi (n = 5); brain 2 dpi (n = 8), 4 dpi (n = 8), 7 dpi (n = 8). 10 4 PFU analysis: lung 7 dpi (n = 4); brain 7 dpi (n = 4).10 6 Serum RT-PCR assay: Sham (n = 6), 2dpi (n = 6), 4 dpi (n = 6), 7 dpi (n = 15). 10 6 Serum infectivity assay: Sham (n = 5), 2dpi (n = 3), 4 dpi (n = 5), 7 dpi (n = 8). One-way or two-way ANOVA. * p ≤ 0.05, ** p ≤ 0.01, **** p ≤ 0.0001. For A–D, shades of blue and red asterisks compare sham group vs. male group, and sham group vs. female group, respectively, with darker shades designated for 10 4 PFU, and brighter shades for 10 6 PFU.
    Sars Cov 2 Spike, supplied by Cell Signaling Technology Inc, 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/sars cov 2 spike/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 spike - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Fatal Neurodissemination and SARS-CoV-2 Tropism in K18-hACE2 Mice Is Only Partially Dependent on hACE2 Expression"

    Article Title: Fatal Neurodissemination and SARS-CoV-2 Tropism in K18-hACE2 Mice Is Only Partially Dependent on hACE2 Expression

    Journal: Viruses

    doi: 10.3390/v14030535

    SARS-CoV-2 caused lethal disease in K18-hACE2 mice irrespective of dose (10 4 vs. 10 6 PFU). K18-hACE2 mice (n = 50) were inoculated intranasally with either 1 × 10 4 or 1 × 10 6 plaque forming units (PFU), with n = 6 additional Sham/PBS negative controls. Body weight ( A ), clinical signs ( B ), body temperature ( C ), and survival ( D ) were monitored daily in Sham/PBS animals (black) and in infected animals (male, red; female, blue; up to 14 dpi). Mice meeting euthanasia criteria were counted dead the following day. Viral loads (viral RNA genome copy numbers/mg of tissue) or viral titers (infectious virus particles; PFU/mg of tissue) were quantified in the lung and brain ( E – G ). RNA copies were also examined in the serum (genome copies/mL) either directly on serum ( H ) or via a re-infectivity assay ( I ) using Vero E6 cells. The limit of detection is shown with a dashed line. Clinical data ( A – D ): 10 4 PFU; male (n = 9), female (n = 6); 10 6 PFU male (19), female (n = 16); Sham/PBS male (n = 3), female (n = 3). Molecular and virologic data ( E – I ). 10 6 RT-PCR: lung 2 dpi (n = 3), 4 dpi (n = 6), 7 dpi (n = 8); brain 2 dpi (n = 3), 4 dpi (n = 5), 7 dpi (n = 7). 10 6 PFU analysis: lung 2 dpi (n = 8), 4 dpi (n = 8), 7 dpi (n = 5); brain 2 dpi (n = 8), 4 dpi (n = 8), 7 dpi (n = 8). 10 4 PFU analysis: lung 7 dpi (n = 4); brain 7 dpi (n = 4).10 6 Serum RT-PCR assay: Sham (n = 6), 2dpi (n = 6), 4 dpi (n = 6), 7 dpi (n = 15). 10 6 Serum infectivity assay: Sham (n = 5), 2dpi (n = 3), 4 dpi (n = 5), 7 dpi (n = 8). One-way or two-way ANOVA. * p ≤ 0.05, ** p ≤ 0.01, **** p ≤ 0.0001. For A–D, shades of blue and red asterisks compare sham group vs. male group, and sham group vs. female group, respectively, with darker shades designated for 10 4 PFU, and brighter shades for 10 6 PFU.
    Figure Legend Snippet: SARS-CoV-2 caused lethal disease in K18-hACE2 mice irrespective of dose (10 4 vs. 10 6 PFU). K18-hACE2 mice (n = 50) were inoculated intranasally with either 1 × 10 4 or 1 × 10 6 plaque forming units (PFU), with n = 6 additional Sham/PBS negative controls. Body weight ( A ), clinical signs ( B ), body temperature ( C ), and survival ( D ) were monitored daily in Sham/PBS animals (black) and in infected animals (male, red; female, blue; up to 14 dpi). Mice meeting euthanasia criteria were counted dead the following day. Viral loads (viral RNA genome copy numbers/mg of tissue) or viral titers (infectious virus particles; PFU/mg of tissue) were quantified in the lung and brain ( E – G ). RNA copies were also examined in the serum (genome copies/mL) either directly on serum ( H ) or via a re-infectivity assay ( I ) using Vero E6 cells. The limit of detection is shown with a dashed line. Clinical data ( A – D ): 10 4 PFU; male (n = 9), female (n = 6); 10 6 PFU male (19), female (n = 16); Sham/PBS male (n = 3), female (n = 3). Molecular and virologic data ( E – I ). 10 6 RT-PCR: lung 2 dpi (n = 3), 4 dpi (n = 6), 7 dpi (n = 8); brain 2 dpi (n = 3), 4 dpi (n = 5), 7 dpi (n = 7). 10 6 PFU analysis: lung 2 dpi (n = 8), 4 dpi (n = 8), 7 dpi (n = 5); brain 2 dpi (n = 8), 4 dpi (n = 8), 7 dpi (n = 8). 10 4 PFU analysis: lung 7 dpi (n = 4); brain 7 dpi (n = 4).10 6 Serum RT-PCR assay: Sham (n = 6), 2dpi (n = 6), 4 dpi (n = 6), 7 dpi (n = 15). 10 6 Serum infectivity assay: Sham (n = 5), 2dpi (n = 3), 4 dpi (n = 5), 7 dpi (n = 8). One-way or two-way ANOVA. * p ≤ 0.05, ** p ≤ 0.01, **** p ≤ 0.0001. For A–D, shades of blue and red asterisks compare sham group vs. male group, and sham group vs. female group, respectively, with darker shades designated for 10 4 PFU, and brighter shades for 10 6 PFU.

    Techniques Used: Infection, Reverse Transcription Polymerase Chain Reaction

    Temporal analysis of SARS-CoV-2 infection in the nasal cavity of K18-hACE2 mice. Histological changes, and viral protein (brown) and RNA (red) distribution and abundance were assessed in non-infected (Sham/PBS: A , D , G , J ; n = 3) and infected mice at 2 ( B , E , H , K , n = 3) and 4 ( C , F , I , L , n = 5) days following intranasal inoculation. At 2 dpi, neutrophilic rhinitis in the rostral and intermediate turbinates ( B , arrow) correlated with intraepithelial SARS-CoV-2 protein and RNA ( E , inset). Viral protein and RNA were detected in the olfactory neuroepithelium (ONE, K and inset) in the absence of histologic lesions ( H ). At 4 dpi, only sporadically infected cells were noted in the epithelium lining the nasal turbinates and ONE ( F , L , arrow, and insets) in the absence of histologic lesions ( C , I ). Sham/PBS-infected are depicted in A , D , G , J . H&E, DAB IHC (viral protein), and Fast Red ISH (viral RNA), 200× total magnification. Bar = 100 μm.
    Figure Legend Snippet: Temporal analysis of SARS-CoV-2 infection in the nasal cavity of K18-hACE2 mice. Histological changes, and viral protein (brown) and RNA (red) distribution and abundance were assessed in non-infected (Sham/PBS: A , D , G , J ; n = 3) and infected mice at 2 ( B , E , H , K , n = 3) and 4 ( C , F , I , L , n = 5) days following intranasal inoculation. At 2 dpi, neutrophilic rhinitis in the rostral and intermediate turbinates ( B , arrow) correlated with intraepithelial SARS-CoV-2 protein and RNA ( E , inset). Viral protein and RNA were detected in the olfactory neuroepithelium (ONE, K and inset) in the absence of histologic lesions ( H ). At 4 dpi, only sporadically infected cells were noted in the epithelium lining the nasal turbinates and ONE ( F , L , arrow, and insets) in the absence of histologic lesions ( C , I ). Sham/PBS-infected are depicted in A , D , G , J . H&E, DAB IHC (viral protein), and Fast Red ISH (viral RNA), 200× total magnification. Bar = 100 μm.

    Techniques Used: Infection

    Temporal qualitative and quantitative analysis of SARS-CoV-2 pneumonia in K18-hACE2 mice. Lung tissues from non-infected mice (PBS/Sham inoculated: ( A , B ) and from infected mice at 2 dpi ( C , D ), 4 dpi ( E , F ), 7 dpi ( G , H ) and 14 dpi (n = 2; I , J ) following intranasal inoculation were analyzed. Subgross histological images of the lungs and corresponding pneumonia classifiers for each timepoint are depicted in panel ( K ) (green = normal; yellow = pneumonia). Mild-to-moderate interstitial pneumonia was evident starting at 2 dpi with frequently reactive blood vessels ( D , arrow). At 7 dpi, alveolar type 2 (AT2) cell hyperplasia was observed ( H , arrows). Residual mild-to-moderate pneumonia was observed in the two male survivors at 14 dpi from the survival curve, with rare sporadic lymphoid aggregates ( J -arrows). H&E, 50× ( A , C , E , G , I ; bar = 500 μm), 200× ( B , D , F , H , J ; bar = 100 μm) and 1× ( K ) total magnification. Pneumonia classifier: PBS/Sham (n = 2), 4 dpi (n = 5), 7 dpi (n = 8), 14 dpi (n = 2). One-way ANOVA; ns, non-significant.
    Figure Legend Snippet: Temporal qualitative and quantitative analysis of SARS-CoV-2 pneumonia in K18-hACE2 mice. Lung tissues from non-infected mice (PBS/Sham inoculated: ( A , B ) and from infected mice at 2 dpi ( C , D ), 4 dpi ( E , F ), 7 dpi ( G , H ) and 14 dpi (n = 2; I , J ) following intranasal inoculation were analyzed. Subgross histological images of the lungs and corresponding pneumonia classifiers for each timepoint are depicted in panel ( K ) (green = normal; yellow = pneumonia). Mild-to-moderate interstitial pneumonia was evident starting at 2 dpi with frequently reactive blood vessels ( D , arrow). At 7 dpi, alveolar type 2 (AT2) cell hyperplasia was observed ( H , arrows). Residual mild-to-moderate pneumonia was observed in the two male survivors at 14 dpi from the survival curve, with rare sporadic lymphoid aggregates ( J -arrows). H&E, 50× ( A , C , E , G , I ; bar = 500 μm), 200× ( B , D , F , H , J ; bar = 100 μm) and 1× ( K ) total magnification. Pneumonia classifier: PBS/Sham (n = 2), 4 dpi (n = 5), 7 dpi (n = 8), 14 dpi (n = 2). One-way ANOVA; ns, non-significant.

    Techniques Used: Infection

    SARS-CoV-2 tropism following intranasal inoculation in K18-hACE2. ( A – C ) At 4 dpi, SARS-CoV-2 (yellow) tropism for RAGE+ alveolar type 1 (AT1, magenta) and scattered SPC+ alveolar type 2 (AT2, red) pneumocytes ( B , C , arrowheads, and arrows, respectively) but not for CD31+ endothelial cells. ( B , C ) represent higher magnification images of the white hashed boxes from ( A ), respectively. At 6 dpi-terminal disease ( D – F ), interpretation of transmission electron microscopy images illustrated virus particles (VPs) bound by vesicles in AT1 ( E ) and AT2 ( F ) cells. AT1 contained abundant caveolae. Another unique feature observed in AT1 cells was the presence of cubic membranes (CuM). AT2 pneumocytes were characterized by presence of lamellar bodies (LB). ( E , F ) represent higher magnification images of the yellow hashed boxes from ( D ). ( G , H ) Viral particles or viral induced membrane alterations were not identified in ciliated or non-ciliated club (Cl) bronchiolar epithelial cells. ( H ) represents a higher magnification of the yellow hashed box in ( G ). Multiplex fluorescent IHC, 100× ( A ; bar = 100 μm) and 200× ( B , C ; bar = 50 μm) total magnification. TEM, bar = 2 μm ( D ), 100 nm ( E – H ), and 3 μm ( G ). A, alveolar lumen; BM, basement membrane; C, capillary; Cav, caveolae; Ci, ciliated epithelium; Cl, club epithelium; CuM, cubic membranes; DMVs, double-membrane vesicles; END, endothelium; VPs, viral particles.
    Figure Legend Snippet: SARS-CoV-2 tropism following intranasal inoculation in K18-hACE2. ( A – C ) At 4 dpi, SARS-CoV-2 (yellow) tropism for RAGE+ alveolar type 1 (AT1, magenta) and scattered SPC+ alveolar type 2 (AT2, red) pneumocytes ( B , C , arrowheads, and arrows, respectively) but not for CD31+ endothelial cells. ( B , C ) represent higher magnification images of the white hashed boxes from ( A ), respectively. At 6 dpi-terminal disease ( D – F ), interpretation of transmission electron microscopy images illustrated virus particles (VPs) bound by vesicles in AT1 ( E ) and AT2 ( F ) cells. AT1 contained abundant caveolae. Another unique feature observed in AT1 cells was the presence of cubic membranes (CuM). AT2 pneumocytes were characterized by presence of lamellar bodies (LB). ( E , F ) represent higher magnification images of the yellow hashed boxes from ( D ). ( G , H ) Viral particles or viral induced membrane alterations were not identified in ciliated or non-ciliated club (Cl) bronchiolar epithelial cells. ( H ) represents a higher magnification of the yellow hashed box in ( G ). Multiplex fluorescent IHC, 100× ( A ; bar = 100 μm) and 200× ( B , C ; bar = 50 μm) total magnification. TEM, bar = 2 μm ( D ), 100 nm ( E – H ), and 3 μm ( G ). A, alveolar lumen; BM, basement membrane; C, capillary; Cav, caveolae; Ci, ciliated epithelium; Cl, club epithelium; CuM, cubic membranes; DMVs, double-membrane vesicles; END, endothelium; VPs, viral particles.

    Techniques Used: Transmission Assay, Electron Microscopy, Multiplex Assay

    Temporal immunoprofiling of the pulmonary host inflammatory response to SARS-CoV-2. ( A – H ) Quantification and 4-plex fluorescent IHC targeting SARS-CoV-2 Spike ( A , E – H ), and macrophage Iba-1+ ( B , E – H ), CD8+ ( C , E – H ) and CD19+ cell ( D , E – H ) infiltration in the lung of PBS/Sham inoculated mice and in SARS-CoV-2 inoculated mice (2 dpi (n = 3), 4 dpi (n = 4), 7 dpi (n = 5) and 14 dpi (n = 2). In inoculated mice, SARS-CoV-2 Spike peaked between 4–7 dpi ( A , F , G ). Iba-1+ macrophages (red) increased significantly peaking at 7 dpi ( B , G ), along with a lower infiltration of CD8+ T lymphocytes-magenta that peaked between 4–7 dpi ( C , F , G ) while Sham/PBS mice had low residual inflammatory cells ( E ). CD19+ B cells arranged in aggregates were only evident in the two male survivors euthanized at 14 dpi ( D , H ). Insets depict immune cell phenotyping outputs that were applied across the whole slide image. Sham/PBS-infected mice (n = 3) were used as baseline controls for quantitative analysis. Multiplex fluorescent IHC ( E – H ): 100× and 400× (insets) total magnification, Bar = 100μm. ( I ) Neutralizing activity of serum isolated from a naïve/non-infected K18-hACE2 (purple) and from the two male14 dpi survivors (survivor 1 and 2, red and black, respectively). An anti-SARS-CoV-2 Spike RBD antibody (anti-RBD, blue) was used as a positive control. Serum was serially diluted by 2-fold. One-way ANOVA. ** p ≤ 0.01; **** p ≤ 0.0001.
    Figure Legend Snippet: Temporal immunoprofiling of the pulmonary host inflammatory response to SARS-CoV-2. ( A – H ) Quantification and 4-plex fluorescent IHC targeting SARS-CoV-2 Spike ( A , E – H ), and macrophage Iba-1+ ( B , E – H ), CD8+ ( C , E – H ) and CD19+ cell ( D , E – H ) infiltration in the lung of PBS/Sham inoculated mice and in SARS-CoV-2 inoculated mice (2 dpi (n = 3), 4 dpi (n = 4), 7 dpi (n = 5) and 14 dpi (n = 2). In inoculated mice, SARS-CoV-2 Spike peaked between 4–7 dpi ( A , F , G ). Iba-1+ macrophages (red) increased significantly peaking at 7 dpi ( B , G ), along with a lower infiltration of CD8+ T lymphocytes-magenta that peaked between 4–7 dpi ( C , F , G ) while Sham/PBS mice had low residual inflammatory cells ( E ). CD19+ B cells arranged in aggregates were only evident in the two male survivors euthanized at 14 dpi ( D , H ). Insets depict immune cell phenotyping outputs that were applied across the whole slide image. Sham/PBS-infected mice (n = 3) were used as baseline controls for quantitative analysis. Multiplex fluorescent IHC ( E – H ): 100× and 400× (insets) total magnification, Bar = 100μm. ( I ) Neutralizing activity of serum isolated from a naïve/non-infected K18-hACE2 (purple) and from the two male14 dpi survivors (survivor 1 and 2, red and black, respectively). An anti-SARS-CoV-2 Spike RBD antibody (anti-RBD, blue) was used as a positive control. Serum was serially diluted by 2-fold. One-way ANOVA. ** p ≤ 0.01; **** p ≤ 0.0001.

    Techniques Used: Infection, Multiplex Assay, Activity Assay, Isolation, Positive Control

    Invasion of SARS-CoV-2 into the central nervous system. ( A ) Sagittal sections of the head of non-infected (Sham/PBS, top panel) and infected (4 and 7 dpi, middle and bottom panel, respectively) were analyzed for viral protein and RNA distribution. At 4 dpi (middle panel), SARS-CoV-2 infected neurons within the mitral layer of the olfactory bulb (1, arrow) as well as small clusters of neuronal bodies within the cerebral cortex (2, SARS-CoV-2 RNA in inset). At 7 dpi (bottom panel), SARS-CoV-2 protein was widespread along the mitral layer of the olfactory bulb (1) and throughout the central nervous system (2, SARS-CoV-2 RNA in inset) with exception of the cerebellum. EPL, external plexiform layer; GCL, granular cell layer; GL, glomerular layer; ML, mitral layer. DAB (viral protein) and Fast Red (viral RNA). 7.5× (bar = 2.5 mm) and 200× (bar = 100 μm) total magnification. On the right of each panel, pictures labelled 1 and 2 are 266× total magnification insets represented by the hashed squares labeled in the lower (7.5×) magnification images. ( B ) Representative three-dimensional profile view (right side) of a K18-hACE2 mouse following inoculation with a SARS-CoV-2 NL virus (10 6 PFU). NanoLuc bioluminescent signal was detected and quantified at 6 dpi following fluorofurimazine injection (Sub-cutaneous) using the InVivoPLOT (InVivoAx) system and an IVIS Spectrum (PerkinElmer) optical imaging instrument. Location of the lungs and brain are indicated.
    Figure Legend Snippet: Invasion of SARS-CoV-2 into the central nervous system. ( A ) Sagittal sections of the head of non-infected (Sham/PBS, top panel) and infected (4 and 7 dpi, middle and bottom panel, respectively) were analyzed for viral protein and RNA distribution. At 4 dpi (middle panel), SARS-CoV-2 infected neurons within the mitral layer of the olfactory bulb (1, arrow) as well as small clusters of neuronal bodies within the cerebral cortex (2, SARS-CoV-2 RNA in inset). At 7 dpi (bottom panel), SARS-CoV-2 protein was widespread along the mitral layer of the olfactory bulb (1) and throughout the central nervous system (2, SARS-CoV-2 RNA in inset) with exception of the cerebellum. EPL, external plexiform layer; GCL, granular cell layer; GL, glomerular layer; ML, mitral layer. DAB (viral protein) and Fast Red (viral RNA). 7.5× (bar = 2.5 mm) and 200× (bar = 100 μm) total magnification. On the right of each panel, pictures labelled 1 and 2 are 266× total magnification insets represented by the hashed squares labeled in the lower (7.5×) magnification images. ( B ) Representative three-dimensional profile view (right side) of a K18-hACE2 mouse following inoculation with a SARS-CoV-2 NL virus (10 6 PFU). NanoLuc bioluminescent signal was detected and quantified at 6 dpi following fluorofurimazine injection (Sub-cutaneous) using the InVivoPLOT (InVivoAx) system and an IVIS Spectrum (PerkinElmer) optical imaging instrument. Location of the lungs and brain are indicated.

    Techniques Used: Infection, Labeling, Injection, Optical Imaging

    SARS-CoV-2-associated neuronal morphological changes, neuronal antigen abundance and glial response. Morphological changes in the brain were noted as early as 6 dpi ( A , B ) and were characterized by variable spongiosis ( B , arrowheads, and top inset) with neuronal degeneration and necrosis ( B , bottom inset and arrows) involving multiple areas within the cerebral cortex and elsewhere. ( C – E ) Quantification of 3-plex fluorescent IHC targeting SARS-CoV-2 Spike protein, astrocytes (GFAP) and microglia (Iba-1) in the brain of Sham/PBS mice and in inoculated mice (2, 4, 7 and 14 dpi). The amount of viral protein rapidly and markedly increased by 7 dpi, along with an intense astrocytic and microglial response ( C – E ). H&E, 100×, bar = 200 μm. Multiplex IHC, 200× total magnification, bar = 100 μm. One-way ANOVA; * p ≤ 0.05.
    Figure Legend Snippet: SARS-CoV-2-associated neuronal morphological changes, neuronal antigen abundance and glial response. Morphological changes in the brain were noted as early as 6 dpi ( A , B ) and were characterized by variable spongiosis ( B , arrowheads, and top inset) with neuronal degeneration and necrosis ( B , bottom inset and arrows) involving multiple areas within the cerebral cortex and elsewhere. ( C – E ) Quantification of 3-plex fluorescent IHC targeting SARS-CoV-2 Spike protein, astrocytes (GFAP) and microglia (Iba-1) in the brain of Sham/PBS mice and in inoculated mice (2, 4, 7 and 14 dpi). The amount of viral protein rapidly and markedly increased by 7 dpi, along with an intense astrocytic and microglial response ( C – E ). H&E, 100×, bar = 200 μm. Multiplex IHC, 200× total magnification, bar = 100 μm. One-way ANOVA; * p ≤ 0.05.

    Techniques Used: Multiplex Assay

    Ultrastructural features of SARS-CoV-2-infected neurons. ( A ) Neurons (Neu) contain abundant intracytoplasmic viral particles and various replication-associated intracytoplasmic membranous structures. Necrotic neurons are diffusely electron dense (NNeu). ( B ) Higher magnification of the squared area on ( A ) depicting double-membrane vesicles (DMVs) and spherules (DMSs). ( C ) Cubic membranes (CMs) are also noted among DMVs and DMSs. ( D ) Higher magnification of the squared area on ( A ). Mature viral particles are indicated as VPs. ( E , F ) VPs are associated with membranous structures related to viral replication (DMVs, DMSs and CMs) and with the rough endoplasmic reticulum (RER). ( G ) Necrotic neurons (NNeu) are diffusely electron dense and contain numerous cytoplasmic vacuoles. ( H ) Higher magnification of squared area on ( G ). Note the high cytoplasmic electron-density. Scale bars = 100 nm.
    Figure Legend Snippet: Ultrastructural features of SARS-CoV-2-infected neurons. ( A ) Neurons (Neu) contain abundant intracytoplasmic viral particles and various replication-associated intracytoplasmic membranous structures. Necrotic neurons are diffusely electron dense (NNeu). ( B ) Higher magnification of the squared area on ( A ) depicting double-membrane vesicles (DMVs) and spherules (DMSs). ( C ) Cubic membranes (CMs) are also noted among DMVs and DMSs. ( D ) Higher magnification of the squared area on ( A ). Mature viral particles are indicated as VPs. ( E , F ) VPs are associated with membranous structures related to viral replication (DMVs, DMSs and CMs) and with the rough endoplasmic reticulum (RER). ( G ) Necrotic neurons (NNeu) are diffusely electron dense and contain numerous cytoplasmic vacuoles. ( H ) Higher magnification of squared area on ( G ). Note the high cytoplasmic electron-density. Scale bars = 100 nm.

    Techniques Used: Infection

    Distribution of ACE2 in lungs, nasal cavity, brain, and olfactory bulb of wild-type C57BL/6J and uninfected and SARS-CoV-2 infected K18-hACE2 mice. Lung ( A – C ), nasal (rostral/intermediate turbinates [R/I]) and olfactory epithelium (ONE) ( D – F ), olfactory bulb ( G – I ) and brain ( J – L ) from non-infected C57BL/6J, and from non-infected and infected K18-hACE2 mice (7 dpi). K18-hACE2 mice were analyzed via immunohistochemistry using a cross-reactive anti-ACE2 antibody. In the lungs ( A – C ), ACE2 expression (brown) was mostly restricted to the apical membrane of bronchiolar epithelial cells with scattered positive AT2 cells (inset arrows). Nasal (rostral/intermediate turbinates [R/I]) and olfactory epithelium (ONE) were devoid of ACE2 in C57BL/6J mice ( D ), but expression was enhanced in K18-hACE2 mice with intense apical expression ( E , F ). ACE2 expression within the olfactory bulb ( G – I ) and the brain ( J – L ) was restricted to capillary endothelium with no neuronal expression. DAB, 200× total magnification. Bar = 100 μm.
    Figure Legend Snippet: Distribution of ACE2 in lungs, nasal cavity, brain, and olfactory bulb of wild-type C57BL/6J and uninfected and SARS-CoV-2 infected K18-hACE2 mice. Lung ( A – C ), nasal (rostral/intermediate turbinates [R/I]) and olfactory epithelium (ONE) ( D – F ), olfactory bulb ( G – I ) and brain ( J – L ) from non-infected C57BL/6J, and from non-infected and infected K18-hACE2 mice (7 dpi). K18-hACE2 mice were analyzed via immunohistochemistry using a cross-reactive anti-ACE2 antibody. In the lungs ( A – C ), ACE2 expression (brown) was mostly restricted to the apical membrane of bronchiolar epithelial cells with scattered positive AT2 cells (inset arrows). Nasal (rostral/intermediate turbinates [R/I]) and olfactory epithelium (ONE) were devoid of ACE2 in C57BL/6J mice ( D ), but expression was enhanced in K18-hACE2 mice with intense apical expression ( E , F ). ACE2 expression within the olfactory bulb ( G – I ) and the brain ( J – L ) was restricted to capillary endothelium with no neuronal expression. DAB, 200× total magnification. Bar = 100 μm.

    Techniques Used: Infection, Immunohistochemistry, Expressing

    mean sd 18 50  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc mean sd 18 50
    Mean Sd 18 50, supplied by Cell Signaling Technology Inc, 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/mean sd 18 50/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mean sd 18 50 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    Cell Signaling Technology Inc sars cov 2 1 2 spike protein 2b3e5
    Specificities and sources of primary and secondary antibodies.
    Sars Cov 2 1 2 Spike Protein 2b3e5, supplied by Cell Signaling Technology Inc, 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/sars cov 2 1 2 spike protein 2b3e5/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 1 2 spike protein 2b3e5 - by Bioz Stars, 2023-01
    94/100 stars
      Buy from Supplier

    94
    Cell Signaling Technology Inc sars cov 2 spike rbd
    Prevalence of IgG specific for betacoronavirus RBDs in study cohort. Plasma was assayed by ELISA using recombinant, multimeric RBD as antigen. (a) <t>SARS-COV-2</t> spike 319-591 with p values from ANOVA with Tukey HSD for all pairwise comparisons shown beneath. (b) SARS-CoV spike 384-655 with p values shown as above, (c) MERS spike 306-577. Each point indicates an individual participant. Blue bars show median values. N = 7–30. (d) Comparison of absorbance values for MERS and SARS-CoV RBD binding IgG for all participants. Each point indicates an individual participant, n = 124. The orange circle indicates participants with detectable amounts of MERS RBD-binding IgG, showing correlation with SARS-CoV-binding IgG in this subset. IgG specific to recombinant, multimeric RBD from seasonal betacoronaviruses, (e) OC43 spike 315-675, (f) HKU1 spike 307-675 assayed by ELISA as above.
    Sars Cov 2 Spike Rbd, supplied by Cell Signaling Technology Inc, 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/sars cov 2 spike rbd/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 spike rbd - by Bioz Stars, 2023-01
    94/100 stars
      Buy from Supplier

    94
    Cell Signaling Technology Inc sars cov 2 spike
    <t>SARS-CoV-2</t> caused lethal disease in K18-hACE2 mice irrespective of dose (10 4 vs. 10 6 PFU). K18-hACE2 mice (n = 50) were inoculated intranasally with either 1 × 10 4 or 1 × 10 6 plaque forming units (PFU), with n = 6 additional Sham/PBS negative controls. Body weight ( A ), clinical signs ( B ), body temperature ( C ), and survival ( D ) were monitored daily in Sham/PBS animals (black) and in infected animals (male, red; female, blue; up to 14 dpi). Mice meeting euthanasia criteria were counted dead the following day. Viral loads (viral RNA genome copy numbers/mg of tissue) or viral titers (infectious virus particles; PFU/mg of tissue) were quantified in the lung and brain ( E – G ). RNA copies were also examined in the serum (genome copies/mL) either directly on serum ( H ) or via a re-infectivity assay ( I ) using Vero E6 cells. The limit of detection is shown with a dashed line. Clinical data ( A – D ): 10 4 PFU; male (n = 9), female (n = 6); 10 6 PFU male (19), female (n = 16); Sham/PBS male (n = 3), female (n = 3). Molecular and virologic data ( E – I ). 10 6 RT-PCR: lung 2 dpi (n = 3), 4 dpi (n = 6), 7 dpi (n = 8); brain 2 dpi (n = 3), 4 dpi (n = 5), 7 dpi (n = 7). 10 6 PFU analysis: lung 2 dpi (n = 8), 4 dpi (n = 8), 7 dpi (n = 5); brain 2 dpi (n = 8), 4 dpi (n = 8), 7 dpi (n = 8). 10 4 PFU analysis: lung 7 dpi (n = 4); brain 7 dpi (n = 4).10 6 Serum RT-PCR assay: Sham (n = 6), 2dpi (n = 6), 4 dpi (n = 6), 7 dpi (n = 15). 10 6 Serum infectivity assay: Sham (n = 5), 2dpi (n = 3), 4 dpi (n = 5), 7 dpi (n = 8). One-way or two-way ANOVA. * p ≤ 0.05, ** p ≤ 0.01, **** p ≤ 0.0001. For A–D, shades of blue and red asterisks compare sham group vs. male group, and sham group vs. female group, respectively, with darker shades designated for 10 4 PFU, and brighter shades for 10 6 PFU.
    Sars Cov 2 Spike, supplied by Cell Signaling Technology Inc, 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/sars cov 2 spike/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 spike - by Bioz Stars, 2023-01
    94/100 stars
      Buy from Supplier

    94
    Cell Signaling Technology Inc mean sd 18 50
    <t>SARS-CoV-2</t> caused lethal disease in K18-hACE2 mice irrespective of dose (10 4 vs. 10 6 PFU). K18-hACE2 mice (n = 50) were inoculated intranasally with either 1 × 10 4 or 1 × 10 6 plaque forming units (PFU), with n = 6 additional Sham/PBS negative controls. Body weight ( A ), clinical signs ( B ), body temperature ( C ), and survival ( D ) were monitored daily in Sham/PBS animals (black) and in infected animals (male, red; female, blue; up to 14 dpi). Mice meeting euthanasia criteria were counted dead the following day. Viral loads (viral RNA genome copy numbers/mg of tissue) or viral titers (infectious virus particles; PFU/mg of tissue) were quantified in the lung and brain ( E – G ). RNA copies were also examined in the serum (genome copies/mL) either directly on serum ( H ) or via a re-infectivity assay ( I ) using Vero E6 cells. The limit of detection is shown with a dashed line. Clinical data ( A – D ): 10 4 PFU; male (n = 9), female (n = 6); 10 6 PFU male (19), female (n = 16); Sham/PBS male (n = 3), female (n = 3). Molecular and virologic data ( E – I ). 10 6 RT-PCR: lung 2 dpi (n = 3), 4 dpi (n = 6), 7 dpi (n = 8); brain 2 dpi (n = 3), 4 dpi (n = 5), 7 dpi (n = 7). 10 6 PFU analysis: lung 2 dpi (n = 8), 4 dpi (n = 8), 7 dpi (n = 5); brain 2 dpi (n = 8), 4 dpi (n = 8), 7 dpi (n = 8). 10 4 PFU analysis: lung 7 dpi (n = 4); brain 7 dpi (n = 4).10 6 Serum RT-PCR assay: Sham (n = 6), 2dpi (n = 6), 4 dpi (n = 6), 7 dpi (n = 15). 10 6 Serum infectivity assay: Sham (n = 5), 2dpi (n = 3), 4 dpi (n = 5), 7 dpi (n = 8). One-way or two-way ANOVA. * p ≤ 0.05, ** p ≤ 0.01, **** p ≤ 0.0001. For A–D, shades of blue and red asterisks compare sham group vs. male group, and sham group vs. female group, respectively, with darker shades designated for 10 4 PFU, and brighter shades for 10 6 PFU.
    Mean Sd 18 50, supplied by Cell Signaling Technology Inc, 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/mean sd 18 50/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mean sd 18 50 - by Bioz Stars, 2023-01
    94/100 stars
      Buy from Supplier

    Image Search Results


    Specificities and sources of primary and secondary antibodies.

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Development of a high-sensitivity and short-duration fluorescence in situ hybridization method for viral mRNA detection in HEK 293T cells

    doi: 10.3389/fcimb.2022.960938

    Figure Lengend Snippet: Specificities and sources of primary and secondary antibodies.

    Article Snippet: SARS-CoV-2 1/2 Spike protein(2B3E5) , , Mouse IgG, monoclonal , Cell Signaling Technology (Danvers, MA, United States).

    Techniques:

    Detection of SARS-CoV-2 E mRNA and E/S protein in HEK 293T cells at 24 h after transfection with SARS-CoV-2 E and S plasmids. (A) SARS-CoV-2 E mRNA (with mixed probes containing BME001 and BME002), SARS-CoV-2 E, and SARS-CoV-2 S are shown in red, green, and blue, respectively. (B) Total mRNAs (detected by Oligo dT probes), SARS-CoV-2 E, and SARS-CoV-2 S are shown in red, green, and blue, respectively. (C) Negative controls by using secondary antibodies. Objective lens: 60×; Scale bar: 10 µm; Nu, nucleus.

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Development of a high-sensitivity and short-duration fluorescence in situ hybridization method for viral mRNA detection in HEK 293T cells

    doi: 10.3389/fcimb.2022.960938

    Figure Lengend Snippet: Detection of SARS-CoV-2 E mRNA and E/S protein in HEK 293T cells at 24 h after transfection with SARS-CoV-2 E and S plasmids. (A) SARS-CoV-2 E mRNA (with mixed probes containing BME001 and BME002), SARS-CoV-2 E, and SARS-CoV-2 S are shown in red, green, and blue, respectively. (B) Total mRNAs (detected by Oligo dT probes), SARS-CoV-2 E, and SARS-CoV-2 S are shown in red, green, and blue, respectively. (C) Negative controls by using secondary antibodies. Objective lens: 60×; Scale bar: 10 µm; Nu, nucleus.

    Article Snippet: SARS-CoV-2 1/2 Spike protein(2B3E5) , , Mouse IgG, monoclonal , Cell Signaling Technology (Danvers, MA, United States).

    Techniques: Transfection

    Prevalence of IgG specific for betacoronavirus RBDs in study cohort. Plasma was assayed by ELISA using recombinant, multimeric RBD as antigen. (a) SARS-COV-2 spike 319-591 with p values from ANOVA with Tukey HSD for all pairwise comparisons shown beneath. (b) SARS-CoV spike 384-655 with p values shown as above, (c) MERS spike 306-577. Each point indicates an individual participant. Blue bars show median values. N = 7–30. (d) Comparison of absorbance values for MERS and SARS-CoV RBD binding IgG for all participants. Each point indicates an individual participant, n = 124. The orange circle indicates participants with detectable amounts of MERS RBD-binding IgG, showing correlation with SARS-CoV-binding IgG in this subset. IgG specific to recombinant, multimeric RBD from seasonal betacoronaviruses, (e) OC43 spike 315-675, (f) HKU1 spike 307-675 assayed by ELISA as above.

    Journal: Virulence

    Article Title: Repertoires of SARS-CoV-2 epitopes targeted by antibodies vary according to severity of COVID-19

    doi: 10.1080/21505594.2022.2073025

    Figure Lengend Snippet: Prevalence of IgG specific for betacoronavirus RBDs in study cohort. Plasma was assayed by ELISA using recombinant, multimeric RBD as antigen. (a) SARS-COV-2 spike 319-591 with p values from ANOVA with Tukey HSD for all pairwise comparisons shown beneath. (b) SARS-CoV spike 384-655 with p values shown as above, (c) MERS spike 306-577. Each point indicates an individual participant. Blue bars show median values. N = 7–30. (d) Comparison of absorbance values for MERS and SARS-CoV RBD binding IgG for all participants. Each point indicates an individual participant, n = 124. The orange circle indicates participants with detectable amounts of MERS RBD-binding IgG, showing correlation with SARS-CoV-binding IgG in this subset. IgG specific to recombinant, multimeric RBD from seasonal betacoronaviruses, (e) OC43 spike 315-675, (f) HKU1 spike 307-675 assayed by ELISA as above.

    Article Snippet: ELISAs were prepared using the following recombinant antigens (expressed in HEK 293 cells with 8 × His tags) coated onto 96-well plates: SARS-CoV-2 Spike RBD (multimeric, 319–591, Cell Signaling Technology #17862), SARS-CoV RBD (multimeric, 306–577), MERS-CoV RBD (multimeric, 364–655), OC43 Spike RBD (multimeric, 315–675), HKU1 Spike RBD (multimeric, 307–675).

    Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Binding Assay

    Prevalence of IgGs specific for SARS-CoV-2 peptides. (b) Individual peptides with significant variation between groups ( p < 0.05, ANOVA, n = 7–30). Bars denote groups with significant differences by pairwise comparison (Tukey HSD, p < 0.05).

    Journal: Virulence

    Article Title: Repertoires of SARS-CoV-2 epitopes targeted by antibodies vary according to severity of COVID-19

    doi: 10.1080/21505594.2022.2073025

    Figure Lengend Snippet: Prevalence of IgGs specific for SARS-CoV-2 peptides. (b) Individual peptides with significant variation between groups ( p < 0.05, ANOVA, n = 7–30). Bars denote groups with significant differences by pairwise comparison (Tukey HSD, p < 0.05).

    Article Snippet: ELISAs were prepared using the following recombinant antigens (expressed in HEK 293 cells with 8 × His tags) coated onto 96-well plates: SARS-CoV-2 Spike RBD (multimeric, 319–591, Cell Signaling Technology #17862), SARS-CoV RBD (multimeric, 306–577), MERS-CoV RBD (multimeric, 364–655), OC43 Spike RBD (multimeric, 315–675), HKU1 Spike RBD (multimeric, 307–675).

    Techniques:

    Comparison between peptide seropositivity frequencies in the primary dataset with an independent cohort. Peptides that showed >20 percentage point difference in frequency of seropositivity between participants in the Chelsea survey who had been hospitalized and those who reported exposure to SARS-CoV-2 are shown in green. The differences in frequency of seropositivity for IgG specific to those peptides between acutely hospitalized patients and people with no history of infection are shown in blue.

    Journal: Virulence

    Article Title: Repertoires of SARS-CoV-2 epitopes targeted by antibodies vary according to severity of COVID-19

    doi: 10.1080/21505594.2022.2073025

    Figure Lengend Snippet: Comparison between peptide seropositivity frequencies in the primary dataset with an independent cohort. Peptides that showed >20 percentage point difference in frequency of seropositivity between participants in the Chelsea survey who had been hospitalized and those who reported exposure to SARS-CoV-2 are shown in green. The differences in frequency of seropositivity for IgG specific to those peptides between acutely hospitalized patients and people with no history of infection are shown in blue.

    Article Snippet: ELISAs were prepared using the following recombinant antigens (expressed in HEK 293 cells with 8 × His tags) coated onto 96-well plates: SARS-CoV-2 Spike RBD (multimeric, 319–591, Cell Signaling Technology #17862), SARS-CoV RBD (multimeric, 306–577), MERS-CoV RBD (multimeric, 364–655), OC43 Spike RBD (multimeric, 315–675), HKU1 Spike RBD (multimeric, 307–675).

    Techniques: Infection

    SARS-CoV-2 caused lethal disease in K18-hACE2 mice irrespective of dose (10 4 vs. 10 6 PFU). K18-hACE2 mice (n = 50) were inoculated intranasally with either 1 × 10 4 or 1 × 10 6 plaque forming units (PFU), with n = 6 additional Sham/PBS negative controls. Body weight ( A ), clinical signs ( B ), body temperature ( C ), and survival ( D ) were monitored daily in Sham/PBS animals (black) and in infected animals (male, red; female, blue; up to 14 dpi). Mice meeting euthanasia criteria were counted dead the following day. Viral loads (viral RNA genome copy numbers/mg of tissue) or viral titers (infectious virus particles; PFU/mg of tissue) were quantified in the lung and brain ( E – G ). RNA copies were also examined in the serum (genome copies/mL) either directly on serum ( H ) or via a re-infectivity assay ( I ) using Vero E6 cells. The limit of detection is shown with a dashed line. Clinical data ( A – D ): 10 4 PFU; male (n = 9), female (n = 6); 10 6 PFU male (19), female (n = 16); Sham/PBS male (n = 3), female (n = 3). Molecular and virologic data ( E – I ). 10 6 RT-PCR: lung 2 dpi (n = 3), 4 dpi (n = 6), 7 dpi (n = 8); brain 2 dpi (n = 3), 4 dpi (n = 5), 7 dpi (n = 7). 10 6 PFU analysis: lung 2 dpi (n = 8), 4 dpi (n = 8), 7 dpi (n = 5); brain 2 dpi (n = 8), 4 dpi (n = 8), 7 dpi (n = 8). 10 4 PFU analysis: lung 7 dpi (n = 4); brain 7 dpi (n = 4).10 6 Serum RT-PCR assay: Sham (n = 6), 2dpi (n = 6), 4 dpi (n = 6), 7 dpi (n = 15). 10 6 Serum infectivity assay: Sham (n = 5), 2dpi (n = 3), 4 dpi (n = 5), 7 dpi (n = 8). One-way or two-way ANOVA. * p ≤ 0.05, ** p ≤ 0.01, **** p ≤ 0.0001. For A–D, shades of blue and red asterisks compare sham group vs. male group, and sham group vs. female group, respectively, with darker shades designated for 10 4 PFU, and brighter shades for 10 6 PFU.

    Journal: Viruses

    Article Title: Fatal Neurodissemination and SARS-CoV-2 Tropism in K18-hACE2 Mice Is Only Partially Dependent on hACE2 Expression

    doi: 10.3390/v14030535

    Figure Lengend Snippet: SARS-CoV-2 caused lethal disease in K18-hACE2 mice irrespective of dose (10 4 vs. 10 6 PFU). K18-hACE2 mice (n = 50) were inoculated intranasally with either 1 × 10 4 or 1 × 10 6 plaque forming units (PFU), with n = 6 additional Sham/PBS negative controls. Body weight ( A ), clinical signs ( B ), body temperature ( C ), and survival ( D ) were monitored daily in Sham/PBS animals (black) and in infected animals (male, red; female, blue; up to 14 dpi). Mice meeting euthanasia criteria were counted dead the following day. Viral loads (viral RNA genome copy numbers/mg of tissue) or viral titers (infectious virus particles; PFU/mg of tissue) were quantified in the lung and brain ( E – G ). RNA copies were also examined in the serum (genome copies/mL) either directly on serum ( H ) or via a re-infectivity assay ( I ) using Vero E6 cells. The limit of detection is shown with a dashed line. Clinical data ( A – D ): 10 4 PFU; male (n = 9), female (n = 6); 10 6 PFU male (19), female (n = 16); Sham/PBS male (n = 3), female (n = 3). Molecular and virologic data ( E – I ). 10 6 RT-PCR: lung 2 dpi (n = 3), 4 dpi (n = 6), 7 dpi (n = 8); brain 2 dpi (n = 3), 4 dpi (n = 5), 7 dpi (n = 7). 10 6 PFU analysis: lung 2 dpi (n = 8), 4 dpi (n = 8), 7 dpi (n = 5); brain 2 dpi (n = 8), 4 dpi (n = 8), 7 dpi (n = 8). 10 4 PFU analysis: lung 7 dpi (n = 4); brain 7 dpi (n = 4).10 6 Serum RT-PCR assay: Sham (n = 6), 2dpi (n = 6), 4 dpi (n = 6), 7 dpi (n = 15). 10 6 Serum infectivity assay: Sham (n = 5), 2dpi (n = 3), 4 dpi (n = 5), 7 dpi (n = 8). One-way or two-way ANOVA. * p ≤ 0.05, ** p ≤ 0.01, **** p ≤ 0.0001. For A–D, shades of blue and red asterisks compare sham group vs. male group, and sham group vs. female group, respectively, with darker shades designated for 10 4 PFU, and brighter shades for 10 6 PFU.

    Article Snippet: Assay 1 , NA , SARS-CoV-2 Spike (S) , Mouse , E7U6O , Cell Signaling Technology (Danvers, MA, USA) , Pre-commercialization , 1:1000 , CC1 (Tris) , DAB.

    Techniques: Infection, Reverse Transcription Polymerase Chain Reaction

    Temporal analysis of SARS-CoV-2 infection in the nasal cavity of K18-hACE2 mice. Histological changes, and viral protein (brown) and RNA (red) distribution and abundance were assessed in non-infected (Sham/PBS: A , D , G , J ; n = 3) and infected mice at 2 ( B , E , H , K , n = 3) and 4 ( C , F , I , L , n = 5) days following intranasal inoculation. At 2 dpi, neutrophilic rhinitis in the rostral and intermediate turbinates ( B , arrow) correlated with intraepithelial SARS-CoV-2 protein and RNA ( E , inset). Viral protein and RNA were detected in the olfactory neuroepithelium (ONE, K and inset) in the absence of histologic lesions ( H ). At 4 dpi, only sporadically infected cells were noted in the epithelium lining the nasal turbinates and ONE ( F , L , arrow, and insets) in the absence of histologic lesions ( C , I ). Sham/PBS-infected are depicted in A , D , G , J . H&E, DAB IHC (viral protein), and Fast Red ISH (viral RNA), 200× total magnification. Bar = 100 μm.

    Journal: Viruses

    Article Title: Fatal Neurodissemination and SARS-CoV-2 Tropism in K18-hACE2 Mice Is Only Partially Dependent on hACE2 Expression

    doi: 10.3390/v14030535

    Figure Lengend Snippet: Temporal analysis of SARS-CoV-2 infection in the nasal cavity of K18-hACE2 mice. Histological changes, and viral protein (brown) and RNA (red) distribution and abundance were assessed in non-infected (Sham/PBS: A , D , G , J ; n = 3) and infected mice at 2 ( B , E , H , K , n = 3) and 4 ( C , F , I , L , n = 5) days following intranasal inoculation. At 2 dpi, neutrophilic rhinitis in the rostral and intermediate turbinates ( B , arrow) correlated with intraepithelial SARS-CoV-2 protein and RNA ( E , inset). Viral protein and RNA were detected in the olfactory neuroepithelium (ONE, K and inset) in the absence of histologic lesions ( H ). At 4 dpi, only sporadically infected cells were noted in the epithelium lining the nasal turbinates and ONE ( F , L , arrow, and insets) in the absence of histologic lesions ( C , I ). Sham/PBS-infected are depicted in A , D , G , J . H&E, DAB IHC (viral protein), and Fast Red ISH (viral RNA), 200× total magnification. Bar = 100 μm.

    Article Snippet: Assay 1 , NA , SARS-CoV-2 Spike (S) , Mouse , E7U6O , Cell Signaling Technology (Danvers, MA, USA) , Pre-commercialization , 1:1000 , CC1 (Tris) , DAB.

    Techniques: Infection

    Temporal qualitative and quantitative analysis of SARS-CoV-2 pneumonia in K18-hACE2 mice. Lung tissues from non-infected mice (PBS/Sham inoculated: ( A , B ) and from infected mice at 2 dpi ( C , D ), 4 dpi ( E , F ), 7 dpi ( G , H ) and 14 dpi (n = 2; I , J ) following intranasal inoculation were analyzed. Subgross histological images of the lungs and corresponding pneumonia classifiers for each timepoint are depicted in panel ( K ) (green = normal; yellow = pneumonia). Mild-to-moderate interstitial pneumonia was evident starting at 2 dpi with frequently reactive blood vessels ( D , arrow). At 7 dpi, alveolar type 2 (AT2) cell hyperplasia was observed ( H , arrows). Residual mild-to-moderate pneumonia was observed in the two male survivors at 14 dpi from the survival curve, with rare sporadic lymphoid aggregates ( J -arrows). H&E, 50× ( A , C , E , G , I ; bar = 500 μm), 200× ( B , D , F , H , J ; bar = 100 μm) and 1× ( K ) total magnification. Pneumonia classifier: PBS/Sham (n = 2), 4 dpi (n = 5), 7 dpi (n = 8), 14 dpi (n = 2). One-way ANOVA; ns, non-significant.

    Journal: Viruses

    Article Title: Fatal Neurodissemination and SARS-CoV-2 Tropism in K18-hACE2 Mice Is Only Partially Dependent on hACE2 Expression

    doi: 10.3390/v14030535

    Figure Lengend Snippet: Temporal qualitative and quantitative analysis of SARS-CoV-2 pneumonia in K18-hACE2 mice. Lung tissues from non-infected mice (PBS/Sham inoculated: ( A , B ) and from infected mice at 2 dpi ( C , D ), 4 dpi ( E , F ), 7 dpi ( G , H ) and 14 dpi (n = 2; I , J ) following intranasal inoculation were analyzed. Subgross histological images of the lungs and corresponding pneumonia classifiers for each timepoint are depicted in panel ( K ) (green = normal; yellow = pneumonia). Mild-to-moderate interstitial pneumonia was evident starting at 2 dpi with frequently reactive blood vessels ( D , arrow). At 7 dpi, alveolar type 2 (AT2) cell hyperplasia was observed ( H , arrows). Residual mild-to-moderate pneumonia was observed in the two male survivors at 14 dpi from the survival curve, with rare sporadic lymphoid aggregates ( J -arrows). H&E, 50× ( A , C , E , G , I ; bar = 500 μm), 200× ( B , D , F , H , J ; bar = 100 μm) and 1× ( K ) total magnification. Pneumonia classifier: PBS/Sham (n = 2), 4 dpi (n = 5), 7 dpi (n = 8), 14 dpi (n = 2). One-way ANOVA; ns, non-significant.

    Article Snippet: Assay 1 , NA , SARS-CoV-2 Spike (S) , Mouse , E7U6O , Cell Signaling Technology (Danvers, MA, USA) , Pre-commercialization , 1:1000 , CC1 (Tris) , DAB.

    Techniques: Infection

    SARS-CoV-2 tropism following intranasal inoculation in K18-hACE2. ( A – C ) At 4 dpi, SARS-CoV-2 (yellow) tropism for RAGE+ alveolar type 1 (AT1, magenta) and scattered SPC+ alveolar type 2 (AT2, red) pneumocytes ( B , C , arrowheads, and arrows, respectively) but not for CD31+ endothelial cells. ( B , C ) represent higher magnification images of the white hashed boxes from ( A ), respectively. At 6 dpi-terminal disease ( D – F ), interpretation of transmission electron microscopy images illustrated virus particles (VPs) bound by vesicles in AT1 ( E ) and AT2 ( F ) cells. AT1 contained abundant caveolae. Another unique feature observed in AT1 cells was the presence of cubic membranes (CuM). AT2 pneumocytes were characterized by presence of lamellar bodies (LB). ( E , F ) represent higher magnification images of the yellow hashed boxes from ( D ). ( G , H ) Viral particles or viral induced membrane alterations were not identified in ciliated or non-ciliated club (Cl) bronchiolar epithelial cells. ( H ) represents a higher magnification of the yellow hashed box in ( G ). Multiplex fluorescent IHC, 100× ( A ; bar = 100 μm) and 200× ( B , C ; bar = 50 μm) total magnification. TEM, bar = 2 μm ( D ), 100 nm ( E – H ), and 3 μm ( G ). A, alveolar lumen; BM, basement membrane; C, capillary; Cav, caveolae; Ci, ciliated epithelium; Cl, club epithelium; CuM, cubic membranes; DMVs, double-membrane vesicles; END, endothelium; VPs, viral particles.

    Journal: Viruses

    Article Title: Fatal Neurodissemination and SARS-CoV-2 Tropism in K18-hACE2 Mice Is Only Partially Dependent on hACE2 Expression

    doi: 10.3390/v14030535

    Figure Lengend Snippet: SARS-CoV-2 tropism following intranasal inoculation in K18-hACE2. ( A – C ) At 4 dpi, SARS-CoV-2 (yellow) tropism for RAGE+ alveolar type 1 (AT1, magenta) and scattered SPC+ alveolar type 2 (AT2, red) pneumocytes ( B , C , arrowheads, and arrows, respectively) but not for CD31+ endothelial cells. ( B , C ) represent higher magnification images of the white hashed boxes from ( A ), respectively. At 6 dpi-terminal disease ( D – F ), interpretation of transmission electron microscopy images illustrated virus particles (VPs) bound by vesicles in AT1 ( E ) and AT2 ( F ) cells. AT1 contained abundant caveolae. Another unique feature observed in AT1 cells was the presence of cubic membranes (CuM). AT2 pneumocytes were characterized by presence of lamellar bodies (LB). ( E , F ) represent higher magnification images of the yellow hashed boxes from ( D ). ( G , H ) Viral particles or viral induced membrane alterations were not identified in ciliated or non-ciliated club (Cl) bronchiolar epithelial cells. ( H ) represents a higher magnification of the yellow hashed box in ( G ). Multiplex fluorescent IHC, 100× ( A ; bar = 100 μm) and 200× ( B , C ; bar = 50 μm) total magnification. TEM, bar = 2 μm ( D ), 100 nm ( E – H ), and 3 μm ( G ). A, alveolar lumen; BM, basement membrane; C, capillary; Cav, caveolae; Ci, ciliated epithelium; Cl, club epithelium; CuM, cubic membranes; DMVs, double-membrane vesicles; END, endothelium; VPs, viral particles.

    Article Snippet: Assay 1 , NA , SARS-CoV-2 Spike (S) , Mouse , E7U6O , Cell Signaling Technology (Danvers, MA, USA) , Pre-commercialization , 1:1000 , CC1 (Tris) , DAB.

    Techniques: Transmission Assay, Electron Microscopy, Multiplex Assay

    Temporal immunoprofiling of the pulmonary host inflammatory response to SARS-CoV-2. ( A – H ) Quantification and 4-plex fluorescent IHC targeting SARS-CoV-2 Spike ( A , E – H ), and macrophage Iba-1+ ( B , E – H ), CD8+ ( C , E – H ) and CD19+ cell ( D , E – H ) infiltration in the lung of PBS/Sham inoculated mice and in SARS-CoV-2 inoculated mice (2 dpi (n = 3), 4 dpi (n = 4), 7 dpi (n = 5) and 14 dpi (n = 2). In inoculated mice, SARS-CoV-2 Spike peaked between 4–7 dpi ( A , F , G ). Iba-1+ macrophages (red) increased significantly peaking at 7 dpi ( B , G ), along with a lower infiltration of CD8+ T lymphocytes-magenta that peaked between 4–7 dpi ( C , F , G ) while Sham/PBS mice had low residual inflammatory cells ( E ). CD19+ B cells arranged in aggregates were only evident in the two male survivors euthanized at 14 dpi ( D , H ). Insets depict immune cell phenotyping outputs that were applied across the whole slide image. Sham/PBS-infected mice (n = 3) were used as baseline controls for quantitative analysis. Multiplex fluorescent IHC ( E – H ): 100× and 400× (insets) total magnification, Bar = 100μm. ( I ) Neutralizing activity of serum isolated from a naïve/non-infected K18-hACE2 (purple) and from the two male14 dpi survivors (survivor 1 and 2, red and black, respectively). An anti-SARS-CoV-2 Spike RBD antibody (anti-RBD, blue) was used as a positive control. Serum was serially diluted by 2-fold. One-way ANOVA. ** p ≤ 0.01; **** p ≤ 0.0001.

    Journal: Viruses

    Article Title: Fatal Neurodissemination and SARS-CoV-2 Tropism in K18-hACE2 Mice Is Only Partially Dependent on hACE2 Expression

    doi: 10.3390/v14030535

    Figure Lengend Snippet: Temporal immunoprofiling of the pulmonary host inflammatory response to SARS-CoV-2. ( A – H ) Quantification and 4-plex fluorescent IHC targeting SARS-CoV-2 Spike ( A , E – H ), and macrophage Iba-1+ ( B , E – H ), CD8+ ( C , E – H ) and CD19+ cell ( D , E – H ) infiltration in the lung of PBS/Sham inoculated mice and in SARS-CoV-2 inoculated mice (2 dpi (n = 3), 4 dpi (n = 4), 7 dpi (n = 5) and 14 dpi (n = 2). In inoculated mice, SARS-CoV-2 Spike peaked between 4–7 dpi ( A , F , G ). Iba-1+ macrophages (red) increased significantly peaking at 7 dpi ( B , G ), along with a lower infiltration of CD8+ T lymphocytes-magenta that peaked between 4–7 dpi ( C , F , G ) while Sham/PBS mice had low residual inflammatory cells ( E ). CD19+ B cells arranged in aggregates were only evident in the two male survivors euthanized at 14 dpi ( D , H ). Insets depict immune cell phenotyping outputs that were applied across the whole slide image. Sham/PBS-infected mice (n = 3) were used as baseline controls for quantitative analysis. Multiplex fluorescent IHC ( E – H ): 100× and 400× (insets) total magnification, Bar = 100μm. ( I ) Neutralizing activity of serum isolated from a naïve/non-infected K18-hACE2 (purple) and from the two male14 dpi survivors (survivor 1 and 2, red and black, respectively). An anti-SARS-CoV-2 Spike RBD antibody (anti-RBD, blue) was used as a positive control. Serum was serially diluted by 2-fold. One-way ANOVA. ** p ≤ 0.01; **** p ≤ 0.0001.

    Article Snippet: Assay 1 , NA , SARS-CoV-2 Spike (S) , Mouse , E7U6O , Cell Signaling Technology (Danvers, MA, USA) , Pre-commercialization , 1:1000 , CC1 (Tris) , DAB.

    Techniques: Infection, Multiplex Assay, Activity Assay, Isolation, Positive Control

    Invasion of SARS-CoV-2 into the central nervous system. ( A ) Sagittal sections of the head of non-infected (Sham/PBS, top panel) and infected (4 and 7 dpi, middle and bottom panel, respectively) were analyzed for viral protein and RNA distribution. At 4 dpi (middle panel), SARS-CoV-2 infected neurons within the mitral layer of the olfactory bulb (1, arrow) as well as small clusters of neuronal bodies within the cerebral cortex (2, SARS-CoV-2 RNA in inset). At 7 dpi (bottom panel), SARS-CoV-2 protein was widespread along the mitral layer of the olfactory bulb (1) and throughout the central nervous system (2, SARS-CoV-2 RNA in inset) with exception of the cerebellum. EPL, external plexiform layer; GCL, granular cell layer; GL, glomerular layer; ML, mitral layer. DAB (viral protein) and Fast Red (viral RNA). 7.5× (bar = 2.5 mm) and 200× (bar = 100 μm) total magnification. On the right of each panel, pictures labelled 1 and 2 are 266× total magnification insets represented by the hashed squares labeled in the lower (7.5×) magnification images. ( B ) Representative three-dimensional profile view (right side) of a K18-hACE2 mouse following inoculation with a SARS-CoV-2 NL virus (10 6 PFU). NanoLuc bioluminescent signal was detected and quantified at 6 dpi following fluorofurimazine injection (Sub-cutaneous) using the InVivoPLOT (InVivoAx) system and an IVIS Spectrum (PerkinElmer) optical imaging instrument. Location of the lungs and brain are indicated.

    Journal: Viruses

    Article Title: Fatal Neurodissemination and SARS-CoV-2 Tropism in K18-hACE2 Mice Is Only Partially Dependent on hACE2 Expression

    doi: 10.3390/v14030535

    Figure Lengend Snippet: Invasion of SARS-CoV-2 into the central nervous system. ( A ) Sagittal sections of the head of non-infected (Sham/PBS, top panel) and infected (4 and 7 dpi, middle and bottom panel, respectively) were analyzed for viral protein and RNA distribution. At 4 dpi (middle panel), SARS-CoV-2 infected neurons within the mitral layer of the olfactory bulb (1, arrow) as well as small clusters of neuronal bodies within the cerebral cortex (2, SARS-CoV-2 RNA in inset). At 7 dpi (bottom panel), SARS-CoV-2 protein was widespread along the mitral layer of the olfactory bulb (1) and throughout the central nervous system (2, SARS-CoV-2 RNA in inset) with exception of the cerebellum. EPL, external plexiform layer; GCL, granular cell layer; GL, glomerular layer; ML, mitral layer. DAB (viral protein) and Fast Red (viral RNA). 7.5× (bar = 2.5 mm) and 200× (bar = 100 μm) total magnification. On the right of each panel, pictures labelled 1 and 2 are 266× total magnification insets represented by the hashed squares labeled in the lower (7.5×) magnification images. ( B ) Representative three-dimensional profile view (right side) of a K18-hACE2 mouse following inoculation with a SARS-CoV-2 NL virus (10 6 PFU). NanoLuc bioluminescent signal was detected and quantified at 6 dpi following fluorofurimazine injection (Sub-cutaneous) using the InVivoPLOT (InVivoAx) system and an IVIS Spectrum (PerkinElmer) optical imaging instrument. Location of the lungs and brain are indicated.

    Article Snippet: Assay 1 , NA , SARS-CoV-2 Spike (S) , Mouse , E7U6O , Cell Signaling Technology (Danvers, MA, USA) , Pre-commercialization , 1:1000 , CC1 (Tris) , DAB.

    Techniques: Infection, Labeling, Injection, Optical Imaging

    SARS-CoV-2-associated neuronal morphological changes, neuronal antigen abundance and glial response. Morphological changes in the brain were noted as early as 6 dpi ( A , B ) and were characterized by variable spongiosis ( B , arrowheads, and top inset) with neuronal degeneration and necrosis ( B , bottom inset and arrows) involving multiple areas within the cerebral cortex and elsewhere. ( C – E ) Quantification of 3-plex fluorescent IHC targeting SARS-CoV-2 Spike protein, astrocytes (GFAP) and microglia (Iba-1) in the brain of Sham/PBS mice and in inoculated mice (2, 4, 7 and 14 dpi). The amount of viral protein rapidly and markedly increased by 7 dpi, along with an intense astrocytic and microglial response ( C – E ). H&E, 100×, bar = 200 μm. Multiplex IHC, 200× total magnification, bar = 100 μm. One-way ANOVA; * p ≤ 0.05.

    Journal: Viruses

    Article Title: Fatal Neurodissemination and SARS-CoV-2 Tropism in K18-hACE2 Mice Is Only Partially Dependent on hACE2 Expression

    doi: 10.3390/v14030535

    Figure Lengend Snippet: SARS-CoV-2-associated neuronal morphological changes, neuronal antigen abundance and glial response. Morphological changes in the brain were noted as early as 6 dpi ( A , B ) and were characterized by variable spongiosis ( B , arrowheads, and top inset) with neuronal degeneration and necrosis ( B , bottom inset and arrows) involving multiple areas within the cerebral cortex and elsewhere. ( C – E ) Quantification of 3-plex fluorescent IHC targeting SARS-CoV-2 Spike protein, astrocytes (GFAP) and microglia (Iba-1) in the brain of Sham/PBS mice and in inoculated mice (2, 4, 7 and 14 dpi). The amount of viral protein rapidly and markedly increased by 7 dpi, along with an intense astrocytic and microglial response ( C – E ). H&E, 100×, bar = 200 μm. Multiplex IHC, 200× total magnification, bar = 100 μm. One-way ANOVA; * p ≤ 0.05.

    Article Snippet: Assay 1 , NA , SARS-CoV-2 Spike (S) , Mouse , E7U6O , Cell Signaling Technology (Danvers, MA, USA) , Pre-commercialization , 1:1000 , CC1 (Tris) , DAB.

    Techniques: Multiplex Assay

    Ultrastructural features of SARS-CoV-2-infected neurons. ( A ) Neurons (Neu) contain abundant intracytoplasmic viral particles and various replication-associated intracytoplasmic membranous structures. Necrotic neurons are diffusely electron dense (NNeu). ( B ) Higher magnification of the squared area on ( A ) depicting double-membrane vesicles (DMVs) and spherules (DMSs). ( C ) Cubic membranes (CMs) are also noted among DMVs and DMSs. ( D ) Higher magnification of the squared area on ( A ). Mature viral particles are indicated as VPs. ( E , F ) VPs are associated with membranous structures related to viral replication (DMVs, DMSs and CMs) and with the rough endoplasmic reticulum (RER). ( G ) Necrotic neurons (NNeu) are diffusely electron dense and contain numerous cytoplasmic vacuoles. ( H ) Higher magnification of squared area on ( G ). Note the high cytoplasmic electron-density. Scale bars = 100 nm.

    Journal: Viruses

    Article Title: Fatal Neurodissemination and SARS-CoV-2 Tropism in K18-hACE2 Mice Is Only Partially Dependent on hACE2 Expression

    doi: 10.3390/v14030535

    Figure Lengend Snippet: Ultrastructural features of SARS-CoV-2-infected neurons. ( A ) Neurons (Neu) contain abundant intracytoplasmic viral particles and various replication-associated intracytoplasmic membranous structures. Necrotic neurons are diffusely electron dense (NNeu). ( B ) Higher magnification of the squared area on ( A ) depicting double-membrane vesicles (DMVs) and spherules (DMSs). ( C ) Cubic membranes (CMs) are also noted among DMVs and DMSs. ( D ) Higher magnification of the squared area on ( A ). Mature viral particles are indicated as VPs. ( E , F ) VPs are associated with membranous structures related to viral replication (DMVs, DMSs and CMs) and with the rough endoplasmic reticulum (RER). ( G ) Necrotic neurons (NNeu) are diffusely electron dense and contain numerous cytoplasmic vacuoles. ( H ) Higher magnification of squared area on ( G ). Note the high cytoplasmic electron-density. Scale bars = 100 nm.

    Article Snippet: Assay 1 , NA , SARS-CoV-2 Spike (S) , Mouse , E7U6O , Cell Signaling Technology (Danvers, MA, USA) , Pre-commercialization , 1:1000 , CC1 (Tris) , DAB.

    Techniques: Infection

    Distribution of ACE2 in lungs, nasal cavity, brain, and olfactory bulb of wild-type C57BL/6J and uninfected and SARS-CoV-2 infected K18-hACE2 mice. Lung ( A – C ), nasal (rostral/intermediate turbinates [R/I]) and olfactory epithelium (ONE) ( D – F ), olfactory bulb ( G – I ) and brain ( J – L ) from non-infected C57BL/6J, and from non-infected and infected K18-hACE2 mice (7 dpi). K18-hACE2 mice were analyzed via immunohistochemistry using a cross-reactive anti-ACE2 antibody. In the lungs ( A – C ), ACE2 expression (brown) was mostly restricted to the apical membrane of bronchiolar epithelial cells with scattered positive AT2 cells (inset arrows). Nasal (rostral/intermediate turbinates [R/I]) and olfactory epithelium (ONE) were devoid of ACE2 in C57BL/6J mice ( D ), but expression was enhanced in K18-hACE2 mice with intense apical expression ( E , F ). ACE2 expression within the olfactory bulb ( G – I ) and the brain ( J – L ) was restricted to capillary endothelium with no neuronal expression. DAB, 200× total magnification. Bar = 100 μm.

    Journal: Viruses

    Article Title: Fatal Neurodissemination and SARS-CoV-2 Tropism in K18-hACE2 Mice Is Only Partially Dependent on hACE2 Expression

    doi: 10.3390/v14030535

    Figure Lengend Snippet: Distribution of ACE2 in lungs, nasal cavity, brain, and olfactory bulb of wild-type C57BL/6J and uninfected and SARS-CoV-2 infected K18-hACE2 mice. Lung ( A – C ), nasal (rostral/intermediate turbinates [R/I]) and olfactory epithelium (ONE) ( D – F ), olfactory bulb ( G – I ) and brain ( J – L ) from non-infected C57BL/6J, and from non-infected and infected K18-hACE2 mice (7 dpi). K18-hACE2 mice were analyzed via immunohistochemistry using a cross-reactive anti-ACE2 antibody. In the lungs ( A – C ), ACE2 expression (brown) was mostly restricted to the apical membrane of bronchiolar epithelial cells with scattered positive AT2 cells (inset arrows). Nasal (rostral/intermediate turbinates [R/I]) and olfactory epithelium (ONE) were devoid of ACE2 in C57BL/6J mice ( D ), but expression was enhanced in K18-hACE2 mice with intense apical expression ( E , F ). ACE2 expression within the olfactory bulb ( G – I ) and the brain ( J – L ) was restricted to capillary endothelium with no neuronal expression. DAB, 200× total magnification. Bar = 100 μm.

    Article Snippet: Assay 1 , NA , SARS-CoV-2 Spike (S) , Mouse , E7U6O , Cell Signaling Technology (Danvers, MA, USA) , Pre-commercialization , 1:1000 , CC1 (Tris) , DAB.

    Techniques: Infection, Immunohistochemistry, Expressing