sars cov 2 s1 domain  (ATCC)


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    ATCC sars cov 2 s1 domain
    Frequency of IgG <t>anti-SARS-CoV-2</t> concentration categories among vaccinated and unvaccinated participants, comparing the percentage of individuals in divisions based on IgG levels every 1000 BAU/ml
    Sars Cov 2 S1 Domain, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "COVID-19 vaccination in healthcare workers: Long-term benefits and protection"

    Article Title: COVID-19 vaccination in healthcare workers: Long-term benefits and protection

    Journal: Central-European Journal of Immunology

    doi: 10.5114/ceji.2023.134250

    Frequency of IgG anti-SARS-CoV-2 concentration categories among vaccinated and unvaccinated participants, comparing the percentage of individuals in divisions based on IgG levels every 1000 BAU/ml
    Figure Legend Snippet: Frequency of IgG anti-SARS-CoV-2 concentration categories among vaccinated and unvaccinated participants, comparing the percentage of individuals in divisions based on IgG levels every 1000 BAU/ml

    Techniques Used: Concentration Assay

    sars cov 2 data  (ATCC)


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    ATCC sars cov 2 data
    Sars Cov 2 Data, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    sars cov 2  (ATCC)


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    ATCC sars cov 2
    Sars Cov 2, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    type sars cov 2  (ATCC)


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    ATCC type sars cov 2
    a Number of clonal families of BCRs found within the different BMPC clusters (diversity) and probability of finding different clonal families by random selection of cells (Simpson diversity index). A clonal family was defined by V and J gene composition and a CDR3 region with <20% Hamming distance in both the heavy and light chains originating from one donor. b Mutation rates in the framework regions FR1-3 (left) and in the CDR1-3 regions (right) of the heavy and light chain rearrangements across BMPC clusters. c Bubble plot of the mutation rates in the framework regions FR1-3 (left) and in the CDR1-3 regions (right) of BCRs per isotype and cluster. Colour scale indicates median mutation rates. Values below or above the scale limits are shown in blue or red, respectively. Bubble sizes correspond to the percentage of cells expressing a defined isotype within the indicated cluster. d Violin plot depicting the mutation rates in the V gene of the BCRs of BMPC per cluster. Statistical significance between clusters is shown in Supplementary Data (two-tailed Mann–Whitney U test). Horizontal lines represent the median mutation rate. Violins are coloured by the z score of CD19 gene expression in each cluster. e Identification of <t>SARS-CoV-2</t> spike-specific and tetanus toxoid-specific public clones (in black) among analysed BMPC. Public clones were defined by exhibiting over 80% CDR3 sequence identity in both heavy and light chains when compared to the BCR of sequenced peripheral blood and bone marrow spike- and tetanus-specific cells from vaccinated individuals (see Supplementary Fig. and Supplementary Data ). f Relative distribution of spike-specific (red) and tetanus-specific (blue) BMPC (depicted in e ) per cluster. g Comparison of mutation rates within the framework regions FR1-3 (left) and the CDR1-3 regions (right) of spike-specific (red) and tetanus-specific (blue) BMPC per isotype. Horizontal lines represent the median mutation rate. Statistics were performed using a two-tailed Mann–Whitney U test. Source data are provided as a Source Data file.
    Type Sars Cov 2, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Recruitment of plasma cells from IL-21-dependent and IL-21-independent immune reactions to the bone marrow"

    Article Title: Recruitment of plasma cells from IL-21-dependent and IL-21-independent immune reactions to the bone marrow

    Journal: Nature Communications

    doi: 10.1038/s41467-024-48570-0

    a Number of clonal families of BCRs found within the different BMPC clusters (diversity) and probability of finding different clonal families by random selection of cells (Simpson diversity index). A clonal family was defined by V and J gene composition and a CDR3 region with <20% Hamming distance in both the heavy and light chains originating from one donor. b Mutation rates in the framework regions FR1-3 (left) and in the CDR1-3 regions (right) of the heavy and light chain rearrangements across BMPC clusters. c Bubble plot of the mutation rates in the framework regions FR1-3 (left) and in the CDR1-3 regions (right) of BCRs per isotype and cluster. Colour scale indicates median mutation rates. Values below or above the scale limits are shown in blue or red, respectively. Bubble sizes correspond to the percentage of cells expressing a defined isotype within the indicated cluster. d Violin plot depicting the mutation rates in the V gene of the BCRs of BMPC per cluster. Statistical significance between clusters is shown in Supplementary Data (two-tailed Mann–Whitney U test). Horizontal lines represent the median mutation rate. Violins are coloured by the z score of CD19 gene expression in each cluster. e Identification of SARS-CoV-2 spike-specific and tetanus toxoid-specific public clones (in black) among analysed BMPC. Public clones were defined by exhibiting over 80% CDR3 sequence identity in both heavy and light chains when compared to the BCR of sequenced peripheral blood and bone marrow spike- and tetanus-specific cells from vaccinated individuals (see Supplementary Fig. and Supplementary Data ). f Relative distribution of spike-specific (red) and tetanus-specific (blue) BMPC (depicted in e ) per cluster. g Comparison of mutation rates within the framework regions FR1-3 (left) and the CDR1-3 regions (right) of spike-specific (red) and tetanus-specific (blue) BMPC per isotype. Horizontal lines represent the median mutation rate. Statistics were performed using a two-tailed Mann–Whitney U test. Source data are provided as a Source Data file.
    Figure Legend Snippet: a Number of clonal families of BCRs found within the different BMPC clusters (diversity) and probability of finding different clonal families by random selection of cells (Simpson diversity index). A clonal family was defined by V and J gene composition and a CDR3 region with <20% Hamming distance in both the heavy and light chains originating from one donor. b Mutation rates in the framework regions FR1-3 (left) and in the CDR1-3 regions (right) of the heavy and light chain rearrangements across BMPC clusters. c Bubble plot of the mutation rates in the framework regions FR1-3 (left) and in the CDR1-3 regions (right) of BCRs per isotype and cluster. Colour scale indicates median mutation rates. Values below or above the scale limits are shown in blue or red, respectively. Bubble sizes correspond to the percentage of cells expressing a defined isotype within the indicated cluster. d Violin plot depicting the mutation rates in the V gene of the BCRs of BMPC per cluster. Statistical significance between clusters is shown in Supplementary Data (two-tailed Mann–Whitney U test). Horizontal lines represent the median mutation rate. Violins are coloured by the z score of CD19 gene expression in each cluster. e Identification of SARS-CoV-2 spike-specific and tetanus toxoid-specific public clones (in black) among analysed BMPC. Public clones were defined by exhibiting over 80% CDR3 sequence identity in both heavy and light chains when compared to the BCR of sequenced peripheral blood and bone marrow spike- and tetanus-specific cells from vaccinated individuals (see Supplementary Fig. and Supplementary Data ). f Relative distribution of spike-specific (red) and tetanus-specific (blue) BMPC (depicted in e ) per cluster. g Comparison of mutation rates within the framework regions FR1-3 (left) and the CDR1-3 regions (right) of spike-specific (red) and tetanus-specific (blue) BMPC per isotype. Horizontal lines represent the median mutation rate. Statistics were performed using a two-tailed Mann–Whitney U test. Source data are provided as a Source Data file.

    Techniques Used: Selection, Mutagenesis, Expressing, Two Tailed Test, MANN-WHITNEY, Clone Assay, Sequencing, Comparison

    a Schematic representation of vaccination, blood collection and sample analysis. Arrows indicate time points when transcriptome and full-length B-cell receptor repertoire sequencing was performed. b UMAP representation of the expression levels of selected genes in 55071 CD27 high CD38 high sorted peripheral blood ASC from 36 healthy individuals after COVID-19 or diphtheria, tetanus, pertussis (DTP) vaccination (see Supplementary Table for information on participants and different vaccination schemes). Colour scale represents the expression level of the indicated genes. c Identification of SARS-CoV-2 spike-specific (red) public clones among analysed ASC. Public clones were defined by displaying >80% CDR3 sequence identity when compared to the BCR of sequenced peripheral blood and bone marrow spike- and tetanus-specific cells from vaccinated individuals (see Supplementary Fig. and Supplementary Data ). The number of public clones found is shown on the UMAP. d Bubble plot of mutation rates in the framework regions (FR1-3, left) and in the CDR regions (CDR1-3, right) of identified spike-specific public clones per isotype identified in time point after COVID-19 vaccination. Colour scale indicates median mutation rates. Values above the scale limits are shown in red. Bubble sizes correspond to the percentage of spike-specific cells expressing a defined isotype within the indicated group. e Density plots of BMPC significantly enriched in gene signatures from peripheral blood ASC isolated at different time points after vaccination as identified by Gene Set Enrichment Analysis (GSEA). Violin plots of the normalised enrichment score (NES) per BMPC cluster are depicted in Supplementary Fig. . Statistical significance between NES scores is shown in Supplementary Data (two-tailed Mann–Whitney U test). Source data are provided as a Source Data file.
    Figure Legend Snippet: a Schematic representation of vaccination, blood collection and sample analysis. Arrows indicate time points when transcriptome and full-length B-cell receptor repertoire sequencing was performed. b UMAP representation of the expression levels of selected genes in 55071 CD27 high CD38 high sorted peripheral blood ASC from 36 healthy individuals after COVID-19 or diphtheria, tetanus, pertussis (DTP) vaccination (see Supplementary Table for information on participants and different vaccination schemes). Colour scale represents the expression level of the indicated genes. c Identification of SARS-CoV-2 spike-specific (red) public clones among analysed ASC. Public clones were defined by displaying >80% CDR3 sequence identity when compared to the BCR of sequenced peripheral blood and bone marrow spike- and tetanus-specific cells from vaccinated individuals (see Supplementary Fig. and Supplementary Data ). The number of public clones found is shown on the UMAP. d Bubble plot of mutation rates in the framework regions (FR1-3, left) and in the CDR regions (CDR1-3, right) of identified spike-specific public clones per isotype identified in time point after COVID-19 vaccination. Colour scale indicates median mutation rates. Values above the scale limits are shown in red. Bubble sizes correspond to the percentage of spike-specific cells expressing a defined isotype within the indicated group. e Density plots of BMPC significantly enriched in gene signatures from peripheral blood ASC isolated at different time points after vaccination as identified by Gene Set Enrichment Analysis (GSEA). Violin plots of the normalised enrichment score (NES) per BMPC cluster are depicted in Supplementary Fig. . Statistical significance between NES scores is shown in Supplementary Data (two-tailed Mann–Whitney U test). Source data are provided as a Source Data file.

    Techniques Used: Sequencing, Expressing, Clone Assay, Mutagenesis, Isolation, Two Tailed Test, MANN-WHITNEY

    a Representative pseudocolour plots of intracellular double-positive SARS-CoV-2 RBD (left) or tetanus toxoid (TT, right) staining in CD38 high CD138 + CD14 − CD3 − live singlet BMPC (gating strategy in Supplementary Fig. ). b – e Each symbol represents one donor/sample (see Supplementary Table ). Filled symbols represent BMPC samples which were also analysed by single-cell sequencing (Fig. ). b Frequencies of RBD-specific and TT-specific BMPC within total BMPC. Horizontal lines indicate the median. Statistics were performed using the one-tailed Mann–Whitney U test. n = 20 BM samples. c Frequencies of CD19 low cells within RBD-specific BMPC (red), TT-specific BMPC (blue) and total BMPC (black). Horizontal lines indicate the median. Statistics were performed using the Kruskal–Wallis tests with Dunn’s correction for multiple comparisons. n = 20 BM samples. d Frequencies of IgG+ (left) and IgA+ (right) cells within RBD-specific BMPC (red), TT-specific BMPC (blue) and total BMPC (black). Horizontal lines indicate the median. Statistics were performed using the Kruskal–Wallis tests with Dunn’s correction for multiple comparisons. n = 20 BM samples. e Correlation between the frequency of CD19 low RBD-specific BMPC and days after 3rd vaccination against SARS-CoV-2. Statistics were performed using one-tailed Spearman’s correlations. n = 11 BM samples. Source data are provided as a Source Data file.
    Figure Legend Snippet: a Representative pseudocolour plots of intracellular double-positive SARS-CoV-2 RBD (left) or tetanus toxoid (TT, right) staining in CD38 high CD138 + CD14 − CD3 − live singlet BMPC (gating strategy in Supplementary Fig. ). b – e Each symbol represents one donor/sample (see Supplementary Table ). Filled symbols represent BMPC samples which were also analysed by single-cell sequencing (Fig. ). b Frequencies of RBD-specific and TT-specific BMPC within total BMPC. Horizontal lines indicate the median. Statistics were performed using the one-tailed Mann–Whitney U test. n = 20 BM samples. c Frequencies of CD19 low cells within RBD-specific BMPC (red), TT-specific BMPC (blue) and total BMPC (black). Horizontal lines indicate the median. Statistics were performed using the Kruskal–Wallis tests with Dunn’s correction for multiple comparisons. n = 20 BM samples. d Frequencies of IgG+ (left) and IgA+ (right) cells within RBD-specific BMPC (red), TT-specific BMPC (blue) and total BMPC (black). Horizontal lines indicate the median. Statistics were performed using the Kruskal–Wallis tests with Dunn’s correction for multiple comparisons. n = 20 BM samples. e Correlation between the frequency of CD19 low RBD-specific BMPC and days after 3rd vaccination against SARS-CoV-2. Statistics were performed using one-tailed Spearman’s correlations. n = 11 BM samples. Source data are provided as a Source Data file.

    Techniques Used: Staining, Sequencing, One-tailed Test, MANN-WHITNEY

    spike protein expressing pcdna3 1 sars cov 2 sδct  (ATCC)


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    ATCC spike protein expressing pcdna3 1 sars cov 2 sδct
    Neutralizing antibody (NAb) titers ( a ), peptide Spike-specific IFN-γ responses ( b ), and <t>SARS-CoV-2</t> N and E genomic and subgenomic RNA levels ( c ) in Long Covid (LC) patients and convalescent controls (CC) after Covid-19 infection. Responses were measured against SARS-CoV-2 WA1/2020, Delta, and Omicron BA.1. Medians (red bars) are shown. Groups comparison was performed using the two-sided Mann-Whitney tests, and P values are shown. ns refers to P > 0.05.
    Spike Protein Expressing Pcdna3 1 Sars Cov 2 Sδct, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Persistent Activation of Chronic Inflammatory Pathways in Long Covid"

    Article Title: Persistent Activation of Chronic Inflammatory Pathways in Long Covid

    Journal: bioRxiv

    doi: 10.1101/2024.05.11.593709

    Neutralizing antibody (NAb) titers ( a ), peptide Spike-specific IFN-γ responses ( b ), and SARS-CoV-2 N and E genomic and subgenomic RNA levels ( c ) in Long Covid (LC) patients and convalescent controls (CC) after Covid-19 infection. Responses were measured against SARS-CoV-2 WA1/2020, Delta, and Omicron BA.1. Medians (red bars) are shown. Groups comparison was performed using the two-sided Mann-Whitney tests, and P values are shown. ns refers to P > 0.05.
    Figure Legend Snippet: Neutralizing antibody (NAb) titers ( a ), peptide Spike-specific IFN-γ responses ( b ), and SARS-CoV-2 N and E genomic and subgenomic RNA levels ( c ) in Long Covid (LC) patients and convalescent controls (CC) after Covid-19 infection. Responses were measured against SARS-CoV-2 WA1/2020, Delta, and Omicron BA.1. Medians (red bars) are shown. Groups comparison was performed using the two-sided Mann-Whitney tests, and P values are shown. ns refers to P > 0.05.

    Techniques Used: Infection, Comparison, MANN-WHITNEY

    sars cov 2  (ATCC)


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    ATCC sars cov 2
    Sars Cov 2, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rose bengal sars cov 2 complex formation  (ATCC)


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    ATCC rose bengal sars cov 2 complex formation
    Rose Bengal Sars Cov 2 Complex Formation, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    sars cov 2 pseudoviruses  (ATCC)


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    ATCC sars cov 2 pseudoviruses
    Sars Cov 2 Pseudoviruses, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    mouse adapted sars cov 2 ma10 strain  (ATCC)


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    ATCC mouse adapted sars cov 2 ma10 strain
    Blood-brain barrier permeability, neuroinflammation and neurobehavioural changes in <t>SARS-CoV-2</t> infected mice. ( A – F ) Immunofluorescent staining for CD3+ T cells (green) in brainstem ( A – C ) and hippocampus ( D – F ) of mock or SARS-CoV-2 infected mice at 5 days post inoculation. Glut-1 (red) used to visualize endothelial cells. CD3+ T cells are more abundant in SARS-CoV-2 infected mice in ( C ) brainstem ( P < 0.01) and ( F ) hippocampus ( P < 0.001), unpaired t -test. Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. Monochromatic images are provided in . ( G – L ) Immunofluorescent staining for fibrinogen (green) and DAPI (blue) in brainstem ( G – I ) and hippocampus ( J – L ) of mock or SARS-CoV-2 infected mice. Fibrinogen was more abundant in ( I ) brainstem ( P < 0.0001) and ( L ) hippocampus ( P < 0.0001) of SARS-CoV-2 infected mice, unpaired t -test. ( M – R ) Immunofluorescent staining for Iba1 (green) and DAPI (blue) in brainstem ( M – O ) and hippocampus ( P – R ) of mock or SARS-CoV-2 infected mice. The white box indicates the region of magnification. Arrowhead ( N' ) indicates a microglial nodule. Significantly greater Iba1+ area in ( O ) brainstem ( P < 0.001) and ( R ) hippocampus ( P < 0.001) in SARS-CoV-2 infected mice, unpaired t -test. ( S ) SARS-CoV-2 infected mice have significantly lower novel object discrimination in the novel object recognition task for learning and memory than do mock-infected mice ( P < 0.001, unpaired t -test). Discrimination index of 50% indicates novel object preference equal to chance. ( T ) Total object exploration time in the novel object recognition task does not differ between SARS-CoV-2 and mock infected mice, supporting that poor discrimination index in SARS-CoV-2 infected mice is not driven by lack of interest in objects or motility to reach objects. ( U ) SARS-CoV-2 infected mice are significantly slower than mock infected mice to complete the pole descent task for complex motor coordination ( P < 0.0001, unpaired t -test). ( V ) Bouts of spontaneous grooming have longer duration in SARS-CoV-2 infected mice ( P < 0.05, unpaired t -test). ( W ) Fewer individual bouts of spontaneous grooming in SARS-CoV-2 infected mice ( P < 0.05, unpaired t -test).
    Mouse Adapted Sars Cov 2 Ma10 Strain, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Engineered Wnt7a ligands rescue blood–brain barrier and cognitive deficits in a COVID-19 mouse model"

    Article Title: Engineered Wnt7a ligands rescue blood–brain barrier and cognitive deficits in a COVID-19 mouse model

    Journal: Brain

    doi: 10.1093/brain/awae031

    Blood-brain barrier permeability, neuroinflammation and neurobehavioural changes in SARS-CoV-2 infected mice. ( A – F ) Immunofluorescent staining for CD3+ T cells (green) in brainstem ( A – C ) and hippocampus ( D – F ) of mock or SARS-CoV-2 infected mice at 5 days post inoculation. Glut-1 (red) used to visualize endothelial cells. CD3+ T cells are more abundant in SARS-CoV-2 infected mice in ( C ) brainstem ( P < 0.01) and ( F ) hippocampus ( P < 0.001), unpaired t -test. Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. Monochromatic images are provided in . ( G – L ) Immunofluorescent staining for fibrinogen (green) and DAPI (blue) in brainstem ( G – I ) and hippocampus ( J – L ) of mock or SARS-CoV-2 infected mice. Fibrinogen was more abundant in ( I ) brainstem ( P < 0.0001) and ( L ) hippocampus ( P < 0.0001) of SARS-CoV-2 infected mice, unpaired t -test. ( M – R ) Immunofluorescent staining for Iba1 (green) and DAPI (blue) in brainstem ( M – O ) and hippocampus ( P – R ) of mock or SARS-CoV-2 infected mice. The white box indicates the region of magnification. Arrowhead ( N' ) indicates a microglial nodule. Significantly greater Iba1+ area in ( O ) brainstem ( P < 0.001) and ( R ) hippocampus ( P < 0.001) in SARS-CoV-2 infected mice, unpaired t -test. ( S ) SARS-CoV-2 infected mice have significantly lower novel object discrimination in the novel object recognition task for learning and memory than do mock-infected mice ( P < 0.001, unpaired t -test). Discrimination index of 50% indicates novel object preference equal to chance. ( T ) Total object exploration time in the novel object recognition task does not differ between SARS-CoV-2 and mock infected mice, supporting that poor discrimination index in SARS-CoV-2 infected mice is not driven by lack of interest in objects or motility to reach objects. ( U ) SARS-CoV-2 infected mice are significantly slower than mock infected mice to complete the pole descent task for complex motor coordination ( P < 0.0001, unpaired t -test). ( V ) Bouts of spontaneous grooming have longer duration in SARS-CoV-2 infected mice ( P < 0.05, unpaired t -test). ( W ) Fewer individual bouts of spontaneous grooming in SARS-CoV-2 infected mice ( P < 0.05, unpaired t -test).
    Figure Legend Snippet: Blood-brain barrier permeability, neuroinflammation and neurobehavioural changes in SARS-CoV-2 infected mice. ( A – F ) Immunofluorescent staining for CD3+ T cells (green) in brainstem ( A – C ) and hippocampus ( D – F ) of mock or SARS-CoV-2 infected mice at 5 days post inoculation. Glut-1 (red) used to visualize endothelial cells. CD3+ T cells are more abundant in SARS-CoV-2 infected mice in ( C ) brainstem ( P < 0.01) and ( F ) hippocampus ( P < 0.001), unpaired t -test. Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. Monochromatic images are provided in . ( G – L ) Immunofluorescent staining for fibrinogen (green) and DAPI (blue) in brainstem ( G – I ) and hippocampus ( J – L ) of mock or SARS-CoV-2 infected mice. Fibrinogen was more abundant in ( I ) brainstem ( P < 0.0001) and ( L ) hippocampus ( P < 0.0001) of SARS-CoV-2 infected mice, unpaired t -test. ( M – R ) Immunofluorescent staining for Iba1 (green) and DAPI (blue) in brainstem ( M – O ) and hippocampus ( P – R ) of mock or SARS-CoV-2 infected mice. The white box indicates the region of magnification. Arrowhead ( N' ) indicates a microglial nodule. Significantly greater Iba1+ area in ( O ) brainstem ( P < 0.001) and ( R ) hippocampus ( P < 0.001) in SARS-CoV-2 infected mice, unpaired t -test. ( S ) SARS-CoV-2 infected mice have significantly lower novel object discrimination in the novel object recognition task for learning and memory than do mock-infected mice ( P < 0.001, unpaired t -test). Discrimination index of 50% indicates novel object preference equal to chance. ( T ) Total object exploration time in the novel object recognition task does not differ between SARS-CoV-2 and mock infected mice, supporting that poor discrimination index in SARS-CoV-2 infected mice is not driven by lack of interest in objects or motility to reach objects. ( U ) SARS-CoV-2 infected mice are significantly slower than mock infected mice to complete the pole descent task for complex motor coordination ( P < 0.0001, unpaired t -test). ( V ) Bouts of spontaneous grooming have longer duration in SARS-CoV-2 infected mice ( P < 0.05, unpaired t -test). ( W ) Fewer individual bouts of spontaneous grooming in SARS-CoV-2 infected mice ( P < 0.05, unpaired t -test).

    Techniques Used: Permeability, Infection, Staining

    Dysregulation of brain endothelial cell Wnt/β-catenin signalling in SARS-CoV-2 infection. ( A ) Schematic depiction of brain microvascular endothelial cell isolation protocol using gradient centrifugation and positive/negative selection 5 days after respiratory inoculation with SARS-CoV-2. ( B ) Flow cytome­­tric analysis of isolated brain endothelial cells demonstrating uniform population for side-scatter (SSC-H) and forward scatter (FSC-H). ( C ) Flow cytometric analysis showing histogram for CD31 out of all cells in B . Ninety-four per cent of cells were positive for the endothelial cell marker CD31. ( D ) Gene ontology (GO) category analysis of differentially expressed genes in brain endothelial cells of mice with SARS-CoV-2 as compared to mock infection. ( E ) Volcano plot in which differentially expressed genes in KEGG Wnt/β-catenin pathway are depicted in red. Non-differentially expressed KEGG Wnt/β-catenin genes are depicted in pink. Grey dots represent transcripts that are not part of the KEGG Wnt/β-catenin pathway. ( F and G ) Immunostaining for the β-catenin transcriptional target Nkd1 (green) in brain sections of SARS-CoV-2 or mock-infected mice. Brain endothelial cells are visualized with Glut-1 (red). Nuclei are visualized with DAPI (blue). Monochromatic images are provided as . ( H and I ) Quantification of density of Nkd1+ endothelial cells (EC) in the ( H ) hippocampus and ( I ) brainstem of SARS-CoV-2 or mock-infected mice. Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. Unpaired Student’s t -test, ** P < 0.01. FDR = false discovery rate; KEGG = Kyoto Encyclopedia of Genes and Genomes.
    Figure Legend Snippet: Dysregulation of brain endothelial cell Wnt/β-catenin signalling in SARS-CoV-2 infection. ( A ) Schematic depiction of brain microvascular endothelial cell isolation protocol using gradient centrifugation and positive/negative selection 5 days after respiratory inoculation with SARS-CoV-2. ( B ) Flow cytome­­tric analysis of isolated brain endothelial cells demonstrating uniform population for side-scatter (SSC-H) and forward scatter (FSC-H). ( C ) Flow cytometric analysis showing histogram for CD31 out of all cells in B . Ninety-four per cent of cells were positive for the endothelial cell marker CD31. ( D ) Gene ontology (GO) category analysis of differentially expressed genes in brain endothelial cells of mice with SARS-CoV-2 as compared to mock infection. ( E ) Volcano plot in which differentially expressed genes in KEGG Wnt/β-catenin pathway are depicted in red. Non-differentially expressed KEGG Wnt/β-catenin genes are depicted in pink. Grey dots represent transcripts that are not part of the KEGG Wnt/β-catenin pathway. ( F and G ) Immunostaining for the β-catenin transcriptional target Nkd1 (green) in brain sections of SARS-CoV-2 or mock-infected mice. Brain endothelial cells are visualized with Glut-1 (red). Nuclei are visualized with DAPI (blue). Monochromatic images are provided as . ( H and I ) Quantification of density of Nkd1+ endothelial cells (EC) in the ( H ) hippocampus and ( I ) brainstem of SARS-CoV-2 or mock-infected mice. Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. Unpaired Student’s t -test, ** P < 0.01. FDR = false discovery rate; KEGG = Kyoto Encyclopedia of Genes and Genomes.

    Techniques Used: Infection, Cell Isolation, Gradient Centrifugation, Selection, Isolation, Marker, Immunostaining

    Cerebrovascular-targeted engineered Wnt7a K190A protects against neurobehavioural impairment, neuroinflammation and blood–brain barrier leakage in SARS-CoV-2 infection . Mice were treated with AAV-PHP.eB vector control or AAV-PHP.eB-Wnt7a K190A 18 days prior to inoculation with SARS-CoV-2. Behavioural assessment and tissue collection was conducted 5 days after SARS-CoV-2 inoculation. ( A – F ) Immunostaining for CD3+ T cells (green) in brainstem ( A – C ) and hippocampus ( D – F ) of SARS-CoV-2 infected mice treated with AAV-PHP.eB-Wnt7a K190A or AAV-PHP.eB vector control. Glut-1 (red) was used to visualize endothelial cells. SARS-CoV-2 infected AAV-PHP.eB-Wnt7a K190A treated mice had significantly lower density of CD3+ T cells than did AAV-PHP.eB vector control treated mice in brainstem ( C ) and in hippocampus ( F ). Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. Unpaired Student’s t -test, **** P < 0.0001. ( G – L ) Immunostaining for fibrinogen (red) in brainstem (G–I) and hippocampus ( J – L ) of SARS-CoV-2 infected mice treated with AAV-PHP.eB-Wnt7a K190A or AAV-PHP.eB vector control. Glut1 (green) was used to visualize endothelial cells. SARS-CoV-2 infected AAV-PHP.eB-Wnt7a K190A treated mice had significantly lower perivascular fibrinogen than did AAV-PHP.eB vector control treated mice in brainstem ( I ; unpaired t -test with Welch’s correction for unequal variance, * P < 0.05) and in hippocampus ( L , unpaired t -test, **** P < 0.0001). Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. ( M – R ) Immunostaining for CD68 (green), Iba1 (red), and DAPI (blue) in brainstem ( M and N ) and hippocampus ( P and Q ) of SARS-CoV-2 infected mice treated with AAV-PHP.eB-Wnt7a K190A or AAV-PHP.eB vector control. ( O and R ) Quantification of percent Iba1 + area that is positive for CD68. SARS-CoV-2 infected AAV-PHP.eB-Wnt7a K190A treated mice had significantly lower CD68/Iba1 than did AAV-PHP.eB vector control treated mice in brainstem ( O ; unpaired t -test, ** P < 0.01) and in hippocampus ( R ; unpaired t -test, ** P < 0.01). Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. Monochromatic images are provided in . ( S ) The novel object recognition test for learning and memory was conducted in mice treated with AAV-PHP.eB-Wnt7a K190A or AAV-PHP.eB vector control without or with SARS-CoV-2 infection. There was a Treatment [ F (1,29) = 13.05, P = 0.0011], SARS-CoV-2 [ F (1,29) = 11.75, P = 0.0018], and Treatment × SARS-CoV-2 interaction [ F (1,29) = 8.397, P = 0.0071] for novel object preference. Post hoc analysis revealed that SARS-CoV-2 infection impaired novel object recognition in mice treated with the vector control (**** P < 0.0001) but not in mice treated with AAV-PHP.eB-Wnt7a K190A ( P = 0.7145). Indeed, SARS-CoV-2 infected mice treated with AAV-PHP.eB-Wnt7a K190A performed significantly better than SARS-CoV-2 infected mice treated with the control vector (**** P < 0.0001). ( T ) Total object exploration was not significantly influenced by Treatment [ F (1,29) = 0.009622, P = 0.9225], SARS-CoV-2 [ F (1,29) = 2.679, P = 0.1125], or Treatment × SARS-CoV-2 interaction [ F (1,29) = 0.4577, P = 0.5041], supporting that the novel object recognition task was not confounded by lack of interest or impaired motility. ( U ) Pole descent latency, a measure of complex motor coordination impairment, was significantly influenced by Treatment [ F (1,30) = 7.855, P = 0.0088], SARS-CoV-2 [ F (1,30) = 12.10, P = 0.0016], and Treatment × SARS-CoV-2 interaction [ F (1,30) = 6.410, P = 0.0168]. Post hoc analysis revealed the difference was driven by impairment in the vector-treated SARS-CoV-2 group as compared to the AAV-PHP.eB-Wnt7a K190A treated group (*** P < 0.001) or the uninfected group (*** P < 0.001). ( V ) SARS-CoV-2 infected mice treated with AAV-PHP.eB-Wnt7a K190A fell off the pole significantly fewer times than those treated with AAV-PHP.eB vector control (Fisher’s exact test, * P = 0.0198).
    Figure Legend Snippet: Cerebrovascular-targeted engineered Wnt7a K190A protects against neurobehavioural impairment, neuroinflammation and blood–brain barrier leakage in SARS-CoV-2 infection . Mice were treated with AAV-PHP.eB vector control or AAV-PHP.eB-Wnt7a K190A 18 days prior to inoculation with SARS-CoV-2. Behavioural assessment and tissue collection was conducted 5 days after SARS-CoV-2 inoculation. ( A – F ) Immunostaining for CD3+ T cells (green) in brainstem ( A – C ) and hippocampus ( D – F ) of SARS-CoV-2 infected mice treated with AAV-PHP.eB-Wnt7a K190A or AAV-PHP.eB vector control. Glut-1 (red) was used to visualize endothelial cells. SARS-CoV-2 infected AAV-PHP.eB-Wnt7a K190A treated mice had significantly lower density of CD3+ T cells than did AAV-PHP.eB vector control treated mice in brainstem ( C ) and in hippocampus ( F ). Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. Unpaired Student’s t -test, **** P < 0.0001. ( G – L ) Immunostaining for fibrinogen (red) in brainstem (G–I) and hippocampus ( J – L ) of SARS-CoV-2 infected mice treated with AAV-PHP.eB-Wnt7a K190A or AAV-PHP.eB vector control. Glut1 (green) was used to visualize endothelial cells. SARS-CoV-2 infected AAV-PHP.eB-Wnt7a K190A treated mice had significantly lower perivascular fibrinogen than did AAV-PHP.eB vector control treated mice in brainstem ( I ; unpaired t -test with Welch’s correction for unequal variance, * P < 0.05) and in hippocampus ( L , unpaired t -test, **** P < 0.0001). Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. ( M – R ) Immunostaining for CD68 (green), Iba1 (red), and DAPI (blue) in brainstem ( M and N ) and hippocampus ( P and Q ) of SARS-CoV-2 infected mice treated with AAV-PHP.eB-Wnt7a K190A or AAV-PHP.eB vector control. ( O and R ) Quantification of percent Iba1 + area that is positive for CD68. SARS-CoV-2 infected AAV-PHP.eB-Wnt7a K190A treated mice had significantly lower CD68/Iba1 than did AAV-PHP.eB vector control treated mice in brainstem ( O ; unpaired t -test, ** P < 0.01) and in hippocampus ( R ; unpaired t -test, ** P < 0.01). Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. Monochromatic images are provided in . ( S ) The novel object recognition test for learning and memory was conducted in mice treated with AAV-PHP.eB-Wnt7a K190A or AAV-PHP.eB vector control without or with SARS-CoV-2 infection. There was a Treatment [ F (1,29) = 13.05, P = 0.0011], SARS-CoV-2 [ F (1,29) = 11.75, P = 0.0018], and Treatment × SARS-CoV-2 interaction [ F (1,29) = 8.397, P = 0.0071] for novel object preference. Post hoc analysis revealed that SARS-CoV-2 infection impaired novel object recognition in mice treated with the vector control (**** P < 0.0001) but not in mice treated with AAV-PHP.eB-Wnt7a K190A ( P = 0.7145). Indeed, SARS-CoV-2 infected mice treated with AAV-PHP.eB-Wnt7a K190A performed significantly better than SARS-CoV-2 infected mice treated with the control vector (**** P < 0.0001). ( T ) Total object exploration was not significantly influenced by Treatment [ F (1,29) = 0.009622, P = 0.9225], SARS-CoV-2 [ F (1,29) = 2.679, P = 0.1125], or Treatment × SARS-CoV-2 interaction [ F (1,29) = 0.4577, P = 0.5041], supporting that the novel object recognition task was not confounded by lack of interest or impaired motility. ( U ) Pole descent latency, a measure of complex motor coordination impairment, was significantly influenced by Treatment [ F (1,30) = 7.855, P = 0.0088], SARS-CoV-2 [ F (1,30) = 12.10, P = 0.0016], and Treatment × SARS-CoV-2 interaction [ F (1,30) = 6.410, P = 0.0168]. Post hoc analysis revealed the difference was driven by impairment in the vector-treated SARS-CoV-2 group as compared to the AAV-PHP.eB-Wnt7a K190A treated group (*** P < 0.001) or the uninfected group (*** P < 0.001). ( V ) SARS-CoV-2 infected mice treated with AAV-PHP.eB-Wnt7a K190A fell off the pole significantly fewer times than those treated with AAV-PHP.eB vector control (Fisher’s exact test, * P = 0.0198).

    Techniques Used: Infection, Plasmid Preparation, Immunostaining

    sars cov 2 variants  (ATCC)


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    ATCC sars cov 2 variants
    Sars Cov 2 Variants, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC sars cov 2 s1 domain
    Frequency of IgG <t>anti-SARS-CoV-2</t> concentration categories among vaccinated and unvaccinated participants, comparing the percentage of individuals in divisions based on IgG levels every 1000 BAU/ml
    Sars Cov 2 S1 Domain, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC sars cov 2 data
    Frequency of IgG <t>anti-SARS-CoV-2</t> concentration categories among vaccinated and unvaccinated participants, comparing the percentage of individuals in divisions based on IgG levels every 1000 BAU/ml
    Sars Cov 2 Data, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC sars cov 2
    Frequency of IgG <t>anti-SARS-CoV-2</t> concentration categories among vaccinated and unvaccinated participants, comparing the percentage of individuals in divisions based on IgG levels every 1000 BAU/ml
    Sars Cov 2, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC type sars cov 2
    a Number of clonal families of BCRs found within the different BMPC clusters (diversity) and probability of finding different clonal families by random selection of cells (Simpson diversity index). A clonal family was defined by V and J gene composition and a CDR3 region with <20% Hamming distance in both the heavy and light chains originating from one donor. b Mutation rates in the framework regions FR1-3 (left) and in the CDR1-3 regions (right) of the heavy and light chain rearrangements across BMPC clusters. c Bubble plot of the mutation rates in the framework regions FR1-3 (left) and in the CDR1-3 regions (right) of BCRs per isotype and cluster. Colour scale indicates median mutation rates. Values below or above the scale limits are shown in blue or red, respectively. Bubble sizes correspond to the percentage of cells expressing a defined isotype within the indicated cluster. d Violin plot depicting the mutation rates in the V gene of the BCRs of BMPC per cluster. Statistical significance between clusters is shown in Supplementary Data (two-tailed Mann–Whitney U test). Horizontal lines represent the median mutation rate. Violins are coloured by the z score of CD19 gene expression in each cluster. e Identification of <t>SARS-CoV-2</t> spike-specific and tetanus toxoid-specific public clones (in black) among analysed BMPC. Public clones were defined by exhibiting over 80% CDR3 sequence identity in both heavy and light chains when compared to the BCR of sequenced peripheral blood and bone marrow spike- and tetanus-specific cells from vaccinated individuals (see Supplementary Fig. and Supplementary Data ). f Relative distribution of spike-specific (red) and tetanus-specific (blue) BMPC (depicted in e ) per cluster. g Comparison of mutation rates within the framework regions FR1-3 (left) and the CDR1-3 regions (right) of spike-specific (red) and tetanus-specific (blue) BMPC per isotype. Horizontal lines represent the median mutation rate. Statistics were performed using a two-tailed Mann–Whitney U test. Source data are provided as a Source Data file.
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    ATCC spike protein expressing pcdna3 1 sars cov 2 sδct
    Neutralizing antibody (NAb) titers ( a ), peptide Spike-specific IFN-γ responses ( b ), and <t>SARS-CoV-2</t> N and E genomic and subgenomic RNA levels ( c ) in Long Covid (LC) patients and convalescent controls (CC) after Covid-19 infection. Responses were measured against SARS-CoV-2 WA1/2020, Delta, and Omicron BA.1. Medians (red bars) are shown. Groups comparison was performed using the two-sided Mann-Whitney tests, and P values are shown. ns refers to P > 0.05.
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    ATCC rose bengal sars cov 2 complex formation
    Neutralizing antibody (NAb) titers ( a ), peptide Spike-specific IFN-γ responses ( b ), and <t>SARS-CoV-2</t> N and E genomic and subgenomic RNA levels ( c ) in Long Covid (LC) patients and convalescent controls (CC) after Covid-19 infection. Responses were measured against SARS-CoV-2 WA1/2020, Delta, and Omicron BA.1. Medians (red bars) are shown. Groups comparison was performed using the two-sided Mann-Whitney tests, and P values are shown. ns refers to P > 0.05.
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    ATCC sars cov 2 pseudoviruses
    Neutralizing antibody (NAb) titers ( a ), peptide Spike-specific IFN-γ responses ( b ), and <t>SARS-CoV-2</t> N and E genomic and subgenomic RNA levels ( c ) in Long Covid (LC) patients and convalescent controls (CC) after Covid-19 infection. Responses were measured against SARS-CoV-2 WA1/2020, Delta, and Omicron BA.1. Medians (red bars) are shown. Groups comparison was performed using the two-sided Mann-Whitney tests, and P values are shown. ns refers to P > 0.05.
    Sars Cov 2 Pseudoviruses, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC mouse adapted sars cov 2 ma10 strain
    Blood-brain barrier permeability, neuroinflammation and neurobehavioural changes in <t>SARS-CoV-2</t> infected mice. ( A – F ) Immunofluorescent staining for CD3+ T cells (green) in brainstem ( A – C ) and hippocampus ( D – F ) of mock or SARS-CoV-2 infected mice at 5 days post inoculation. Glut-1 (red) used to visualize endothelial cells. CD3+ T cells are more abundant in SARS-CoV-2 infected mice in ( C ) brainstem ( P < 0.01) and ( F ) hippocampus ( P < 0.001), unpaired t -test. Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. Monochromatic images are provided in . ( G – L ) Immunofluorescent staining for fibrinogen (green) and DAPI (blue) in brainstem ( G – I ) and hippocampus ( J – L ) of mock or SARS-CoV-2 infected mice. Fibrinogen was more abundant in ( I ) brainstem ( P < 0.0001) and ( L ) hippocampus ( P < 0.0001) of SARS-CoV-2 infected mice, unpaired t -test. ( M – R ) Immunofluorescent staining for Iba1 (green) and DAPI (blue) in brainstem ( M – O ) and hippocampus ( P – R ) of mock or SARS-CoV-2 infected mice. The white box indicates the region of magnification. Arrowhead ( N' ) indicates a microglial nodule. Significantly greater Iba1+ area in ( O ) brainstem ( P < 0.001) and ( R ) hippocampus ( P < 0.001) in SARS-CoV-2 infected mice, unpaired t -test. ( S ) SARS-CoV-2 infected mice have significantly lower novel object discrimination in the novel object recognition task for learning and memory than do mock-infected mice ( P < 0.001, unpaired t -test). Discrimination index of 50% indicates novel object preference equal to chance. ( T ) Total object exploration time in the novel object recognition task does not differ between SARS-CoV-2 and mock infected mice, supporting that poor discrimination index in SARS-CoV-2 infected mice is not driven by lack of interest in objects or motility to reach objects. ( U ) SARS-CoV-2 infected mice are significantly slower than mock infected mice to complete the pole descent task for complex motor coordination ( P < 0.0001, unpaired t -test). ( V ) Bouts of spontaneous grooming have longer duration in SARS-CoV-2 infected mice ( P < 0.05, unpaired t -test). ( W ) Fewer individual bouts of spontaneous grooming in SARS-CoV-2 infected mice ( P < 0.05, unpaired t -test).
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    ATCC sars cov 2 variants
    Blood-brain barrier permeability, neuroinflammation and neurobehavioural changes in <t>SARS-CoV-2</t> infected mice. ( A – F ) Immunofluorescent staining for CD3+ T cells (green) in brainstem ( A – C ) and hippocampus ( D – F ) of mock or SARS-CoV-2 infected mice at 5 days post inoculation. Glut-1 (red) used to visualize endothelial cells. CD3+ T cells are more abundant in SARS-CoV-2 infected mice in ( C ) brainstem ( P < 0.01) and ( F ) hippocampus ( P < 0.001), unpaired t -test. Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. Monochromatic images are provided in . ( G – L ) Immunofluorescent staining for fibrinogen (green) and DAPI (blue) in brainstem ( G – I ) and hippocampus ( J – L ) of mock or SARS-CoV-2 infected mice. Fibrinogen was more abundant in ( I ) brainstem ( P < 0.0001) and ( L ) hippocampus ( P < 0.0001) of SARS-CoV-2 infected mice, unpaired t -test. ( M – R ) Immunofluorescent staining for Iba1 (green) and DAPI (blue) in brainstem ( M – O ) and hippocampus ( P – R ) of mock or SARS-CoV-2 infected mice. The white box indicates the region of magnification. Arrowhead ( N' ) indicates a microglial nodule. Significantly greater Iba1+ area in ( O ) brainstem ( P < 0.001) and ( R ) hippocampus ( P < 0.001) in SARS-CoV-2 infected mice, unpaired t -test. ( S ) SARS-CoV-2 infected mice have significantly lower novel object discrimination in the novel object recognition task for learning and memory than do mock-infected mice ( P < 0.001, unpaired t -test). Discrimination index of 50% indicates novel object preference equal to chance. ( T ) Total object exploration time in the novel object recognition task does not differ between SARS-CoV-2 and mock infected mice, supporting that poor discrimination index in SARS-CoV-2 infected mice is not driven by lack of interest in objects or motility to reach objects. ( U ) SARS-CoV-2 infected mice are significantly slower than mock infected mice to complete the pole descent task for complex motor coordination ( P < 0.0001, unpaired t -test). ( V ) Bouts of spontaneous grooming have longer duration in SARS-CoV-2 infected mice ( P < 0.05, unpaired t -test). ( W ) Fewer individual bouts of spontaneous grooming in SARS-CoV-2 infected mice ( P < 0.05, unpaired t -test).
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    Frequency of IgG anti-SARS-CoV-2 concentration categories among vaccinated and unvaccinated participants, comparing the percentage of individuals in divisions based on IgG levels every 1000 BAU/ml

    Journal: Central-European Journal of Immunology

    Article Title: COVID-19 vaccination in healthcare workers: Long-term benefits and protection

    doi: 10.5114/ceji.2023.134250

    Figure Lengend Snippet: Frequency of IgG anti-SARS-CoV-2 concentration categories among vaccinated and unvaccinated participants, comparing the percentage of individuals in divisions based on IgG levels every 1000 BAU/ml

    Article Snippet: The tests were performed in 96-well microplates coated with the SARS-CoV-2 S1 domain (including RBD, receptor-binding domain) recombinantly expressed in the human cell line HEK293 (ATCC).

    Techniques: Concentration Assay

    a Number of clonal families of BCRs found within the different BMPC clusters (diversity) and probability of finding different clonal families by random selection of cells (Simpson diversity index). A clonal family was defined by V and J gene composition and a CDR3 region with <20% Hamming distance in both the heavy and light chains originating from one donor. b Mutation rates in the framework regions FR1-3 (left) and in the CDR1-3 regions (right) of the heavy and light chain rearrangements across BMPC clusters. c Bubble plot of the mutation rates in the framework regions FR1-3 (left) and in the CDR1-3 regions (right) of BCRs per isotype and cluster. Colour scale indicates median mutation rates. Values below or above the scale limits are shown in blue or red, respectively. Bubble sizes correspond to the percentage of cells expressing a defined isotype within the indicated cluster. d Violin plot depicting the mutation rates in the V gene of the BCRs of BMPC per cluster. Statistical significance between clusters is shown in Supplementary Data (two-tailed Mann–Whitney U test). Horizontal lines represent the median mutation rate. Violins are coloured by the z score of CD19 gene expression in each cluster. e Identification of SARS-CoV-2 spike-specific and tetanus toxoid-specific public clones (in black) among analysed BMPC. Public clones were defined by exhibiting over 80% CDR3 sequence identity in both heavy and light chains when compared to the BCR of sequenced peripheral blood and bone marrow spike- and tetanus-specific cells from vaccinated individuals (see Supplementary Fig. and Supplementary Data ). f Relative distribution of spike-specific (red) and tetanus-specific (blue) BMPC (depicted in e ) per cluster. g Comparison of mutation rates within the framework regions FR1-3 (left) and the CDR1-3 regions (right) of spike-specific (red) and tetanus-specific (blue) BMPC per isotype. Horizontal lines represent the median mutation rate. Statistics were performed using a two-tailed Mann–Whitney U test. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: Recruitment of plasma cells from IL-21-dependent and IL-21-independent immune reactions to the bone marrow

    doi: 10.1038/s41467-024-48570-0

    Figure Lengend Snippet: a Number of clonal families of BCRs found within the different BMPC clusters (diversity) and probability of finding different clonal families by random selection of cells (Simpson diversity index). A clonal family was defined by V and J gene composition and a CDR3 region with <20% Hamming distance in both the heavy and light chains originating from one donor. b Mutation rates in the framework regions FR1-3 (left) and in the CDR1-3 regions (right) of the heavy and light chain rearrangements across BMPC clusters. c Bubble plot of the mutation rates in the framework regions FR1-3 (left) and in the CDR1-3 regions (right) of BCRs per isotype and cluster. Colour scale indicates median mutation rates. Values below or above the scale limits are shown in blue or red, respectively. Bubble sizes correspond to the percentage of cells expressing a defined isotype within the indicated cluster. d Violin plot depicting the mutation rates in the V gene of the BCRs of BMPC per cluster. Statistical significance between clusters is shown in Supplementary Data (two-tailed Mann–Whitney U test). Horizontal lines represent the median mutation rate. Violins are coloured by the z score of CD19 gene expression in each cluster. e Identification of SARS-CoV-2 spike-specific and tetanus toxoid-specific public clones (in black) among analysed BMPC. Public clones were defined by exhibiting over 80% CDR3 sequence identity in both heavy and light chains when compared to the BCR of sequenced peripheral blood and bone marrow spike- and tetanus-specific cells from vaccinated individuals (see Supplementary Fig. and Supplementary Data ). f Relative distribution of spike-specific (red) and tetanus-specific (blue) BMPC (depicted in e ) per cluster. g Comparison of mutation rates within the framework regions FR1-3 (left) and the CDR1-3 regions (right) of spike-specific (red) and tetanus-specific (blue) BMPC per isotype. Horizontal lines represent the median mutation rate. Statistics were performed using a two-tailed Mann–Whitney U test. Source data are provided as a Source Data file.

    Article Snippet: HEK293T cells (ATCC CRL-3216) were transfected with a plasmid expressing wild-type SARS-CoV-2 S protein.

    Techniques: Selection, Mutagenesis, Expressing, Two Tailed Test, MANN-WHITNEY, Clone Assay, Sequencing, Comparison

    a Schematic representation of vaccination, blood collection and sample analysis. Arrows indicate time points when transcriptome and full-length B-cell receptor repertoire sequencing was performed. b UMAP representation of the expression levels of selected genes in 55071 CD27 high CD38 high sorted peripheral blood ASC from 36 healthy individuals after COVID-19 or diphtheria, tetanus, pertussis (DTP) vaccination (see Supplementary Table for information on participants and different vaccination schemes). Colour scale represents the expression level of the indicated genes. c Identification of SARS-CoV-2 spike-specific (red) public clones among analysed ASC. Public clones were defined by displaying >80% CDR3 sequence identity when compared to the BCR of sequenced peripheral blood and bone marrow spike- and tetanus-specific cells from vaccinated individuals (see Supplementary Fig. and Supplementary Data ). The number of public clones found is shown on the UMAP. d Bubble plot of mutation rates in the framework regions (FR1-3, left) and in the CDR regions (CDR1-3, right) of identified spike-specific public clones per isotype identified in time point after COVID-19 vaccination. Colour scale indicates median mutation rates. Values above the scale limits are shown in red. Bubble sizes correspond to the percentage of spike-specific cells expressing a defined isotype within the indicated group. e Density plots of BMPC significantly enriched in gene signatures from peripheral blood ASC isolated at different time points after vaccination as identified by Gene Set Enrichment Analysis (GSEA). Violin plots of the normalised enrichment score (NES) per BMPC cluster are depicted in Supplementary Fig. . Statistical significance between NES scores is shown in Supplementary Data (two-tailed Mann–Whitney U test). Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: Recruitment of plasma cells from IL-21-dependent and IL-21-independent immune reactions to the bone marrow

    doi: 10.1038/s41467-024-48570-0

    Figure Lengend Snippet: a Schematic representation of vaccination, blood collection and sample analysis. Arrows indicate time points when transcriptome and full-length B-cell receptor repertoire sequencing was performed. b UMAP representation of the expression levels of selected genes in 55071 CD27 high CD38 high sorted peripheral blood ASC from 36 healthy individuals after COVID-19 or diphtheria, tetanus, pertussis (DTP) vaccination (see Supplementary Table for information on participants and different vaccination schemes). Colour scale represents the expression level of the indicated genes. c Identification of SARS-CoV-2 spike-specific (red) public clones among analysed ASC. Public clones were defined by displaying >80% CDR3 sequence identity when compared to the BCR of sequenced peripheral blood and bone marrow spike- and tetanus-specific cells from vaccinated individuals (see Supplementary Fig. and Supplementary Data ). The number of public clones found is shown on the UMAP. d Bubble plot of mutation rates in the framework regions (FR1-3, left) and in the CDR regions (CDR1-3, right) of identified spike-specific public clones per isotype identified in time point after COVID-19 vaccination. Colour scale indicates median mutation rates. Values above the scale limits are shown in red. Bubble sizes correspond to the percentage of spike-specific cells expressing a defined isotype within the indicated group. e Density plots of BMPC significantly enriched in gene signatures from peripheral blood ASC isolated at different time points after vaccination as identified by Gene Set Enrichment Analysis (GSEA). Violin plots of the normalised enrichment score (NES) per BMPC cluster are depicted in Supplementary Fig. . Statistical significance between NES scores is shown in Supplementary Data (two-tailed Mann–Whitney U test). Source data are provided as a Source Data file.

    Article Snippet: HEK293T cells (ATCC CRL-3216) were transfected with a plasmid expressing wild-type SARS-CoV-2 S protein.

    Techniques: Sequencing, Expressing, Clone Assay, Mutagenesis, Isolation, Two Tailed Test, MANN-WHITNEY

    a Representative pseudocolour plots of intracellular double-positive SARS-CoV-2 RBD (left) or tetanus toxoid (TT, right) staining in CD38 high CD138 + CD14 − CD3 − live singlet BMPC (gating strategy in Supplementary Fig. ). b – e Each symbol represents one donor/sample (see Supplementary Table ). Filled symbols represent BMPC samples which were also analysed by single-cell sequencing (Fig. ). b Frequencies of RBD-specific and TT-specific BMPC within total BMPC. Horizontal lines indicate the median. Statistics were performed using the one-tailed Mann–Whitney U test. n = 20 BM samples. c Frequencies of CD19 low cells within RBD-specific BMPC (red), TT-specific BMPC (blue) and total BMPC (black). Horizontal lines indicate the median. Statistics were performed using the Kruskal–Wallis tests with Dunn’s correction for multiple comparisons. n = 20 BM samples. d Frequencies of IgG+ (left) and IgA+ (right) cells within RBD-specific BMPC (red), TT-specific BMPC (blue) and total BMPC (black). Horizontal lines indicate the median. Statistics were performed using the Kruskal–Wallis tests with Dunn’s correction for multiple comparisons. n = 20 BM samples. e Correlation between the frequency of CD19 low RBD-specific BMPC and days after 3rd vaccination against SARS-CoV-2. Statistics were performed using one-tailed Spearman’s correlations. n = 11 BM samples. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: Recruitment of plasma cells from IL-21-dependent and IL-21-independent immune reactions to the bone marrow

    doi: 10.1038/s41467-024-48570-0

    Figure Lengend Snippet: a Representative pseudocolour plots of intracellular double-positive SARS-CoV-2 RBD (left) or tetanus toxoid (TT, right) staining in CD38 high CD138 + CD14 − CD3 − live singlet BMPC (gating strategy in Supplementary Fig. ). b – e Each symbol represents one donor/sample (see Supplementary Table ). Filled symbols represent BMPC samples which were also analysed by single-cell sequencing (Fig. ). b Frequencies of RBD-specific and TT-specific BMPC within total BMPC. Horizontal lines indicate the median. Statistics were performed using the one-tailed Mann–Whitney U test. n = 20 BM samples. c Frequencies of CD19 low cells within RBD-specific BMPC (red), TT-specific BMPC (blue) and total BMPC (black). Horizontal lines indicate the median. Statistics were performed using the Kruskal–Wallis tests with Dunn’s correction for multiple comparisons. n = 20 BM samples. d Frequencies of IgG+ (left) and IgA+ (right) cells within RBD-specific BMPC (red), TT-specific BMPC (blue) and total BMPC (black). Horizontal lines indicate the median. Statistics were performed using the Kruskal–Wallis tests with Dunn’s correction for multiple comparisons. n = 20 BM samples. e Correlation between the frequency of CD19 low RBD-specific BMPC and days after 3rd vaccination against SARS-CoV-2. Statistics were performed using one-tailed Spearman’s correlations. n = 11 BM samples. Source data are provided as a Source Data file.

    Article Snippet: HEK293T cells (ATCC CRL-3216) were transfected with a plasmid expressing wild-type SARS-CoV-2 S protein.

    Techniques: Staining, Sequencing, One-tailed Test, MANN-WHITNEY

    Neutralizing antibody (NAb) titers ( a ), peptide Spike-specific IFN-γ responses ( b ), and SARS-CoV-2 N and E genomic and subgenomic RNA levels ( c ) in Long Covid (LC) patients and convalescent controls (CC) after Covid-19 infection. Responses were measured against SARS-CoV-2 WA1/2020, Delta, and Omicron BA.1. Medians (red bars) are shown. Groups comparison was performed using the two-sided Mann-Whitney tests, and P values are shown. ns refers to P > 0.05.

    Journal: bioRxiv

    Article Title: Persistent Activation of Chronic Inflammatory Pathways in Long Covid

    doi: 10.1101/2024.05.11.593709

    Figure Lengend Snippet: Neutralizing antibody (NAb) titers ( a ), peptide Spike-specific IFN-γ responses ( b ), and SARS-CoV-2 N and E genomic and subgenomic RNA levels ( c ) in Long Covid (LC) patients and convalescent controls (CC) after Covid-19 infection. Responses were measured against SARS-CoV-2 WA1/2020, Delta, and Omicron BA.1. Medians (red bars) are shown. Groups comparison was performed using the two-sided Mann-Whitney tests, and P values are shown. ns refers to P > 0.05.

    Article Snippet: In brief, a luciferase reporter plasmid pLenti-CMV Puro-Luc (Addgene), packaging construct psPAX2 (AIDS Resource and Reagent Program), and Spike protein expressing pcDNA3.1-SARS-CoV-2 SΔCT were co-transfected into human embryonic kidney (HEK)293 T cells (ATCC CRL_3216) with lipofectamine 2000 (ThermoFisher Scientific).

    Techniques: Infection, Comparison, MANN-WHITNEY

    Blood-brain barrier permeability, neuroinflammation and neurobehavioural changes in SARS-CoV-2 infected mice. ( A – F ) Immunofluorescent staining for CD3+ T cells (green) in brainstem ( A – C ) and hippocampus ( D – F ) of mock or SARS-CoV-2 infected mice at 5 days post inoculation. Glut-1 (red) used to visualize endothelial cells. CD3+ T cells are more abundant in SARS-CoV-2 infected mice in ( C ) brainstem ( P < 0.01) and ( F ) hippocampus ( P < 0.001), unpaired t -test. Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. Monochromatic images are provided in . ( G – L ) Immunofluorescent staining for fibrinogen (green) and DAPI (blue) in brainstem ( G – I ) and hippocampus ( J – L ) of mock or SARS-CoV-2 infected mice. Fibrinogen was more abundant in ( I ) brainstem ( P < 0.0001) and ( L ) hippocampus ( P < 0.0001) of SARS-CoV-2 infected mice, unpaired t -test. ( M – R ) Immunofluorescent staining for Iba1 (green) and DAPI (blue) in brainstem ( M – O ) and hippocampus ( P – R ) of mock or SARS-CoV-2 infected mice. The white box indicates the region of magnification. Arrowhead ( N' ) indicates a microglial nodule. Significantly greater Iba1+ area in ( O ) brainstem ( P < 0.001) and ( R ) hippocampus ( P < 0.001) in SARS-CoV-2 infected mice, unpaired t -test. ( S ) SARS-CoV-2 infected mice have significantly lower novel object discrimination in the novel object recognition task for learning and memory than do mock-infected mice ( P < 0.001, unpaired t -test). Discrimination index of 50% indicates novel object preference equal to chance. ( T ) Total object exploration time in the novel object recognition task does not differ between SARS-CoV-2 and mock infected mice, supporting that poor discrimination index in SARS-CoV-2 infected mice is not driven by lack of interest in objects or motility to reach objects. ( U ) SARS-CoV-2 infected mice are significantly slower than mock infected mice to complete the pole descent task for complex motor coordination ( P < 0.0001, unpaired t -test). ( V ) Bouts of spontaneous grooming have longer duration in SARS-CoV-2 infected mice ( P < 0.05, unpaired t -test). ( W ) Fewer individual bouts of spontaneous grooming in SARS-CoV-2 infected mice ( P < 0.05, unpaired t -test).

    Journal: Brain

    Article Title: Engineered Wnt7a ligands rescue blood–brain barrier and cognitive deficits in a COVID-19 mouse model

    doi: 10.1093/brain/awae031

    Figure Lengend Snippet: Blood-brain barrier permeability, neuroinflammation and neurobehavioural changes in SARS-CoV-2 infected mice. ( A – F ) Immunofluorescent staining for CD3+ T cells (green) in brainstem ( A – C ) and hippocampus ( D – F ) of mock or SARS-CoV-2 infected mice at 5 days post inoculation. Glut-1 (red) used to visualize endothelial cells. CD3+ T cells are more abundant in SARS-CoV-2 infected mice in ( C ) brainstem ( P < 0.01) and ( F ) hippocampus ( P < 0.001), unpaired t -test. Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. Monochromatic images are provided in . ( G – L ) Immunofluorescent staining for fibrinogen (green) and DAPI (blue) in brainstem ( G – I ) and hippocampus ( J – L ) of mock or SARS-CoV-2 infected mice. Fibrinogen was more abundant in ( I ) brainstem ( P < 0.0001) and ( L ) hippocampus ( P < 0.0001) of SARS-CoV-2 infected mice, unpaired t -test. ( M – R ) Immunofluorescent staining for Iba1 (green) and DAPI (blue) in brainstem ( M – O ) and hippocampus ( P – R ) of mock or SARS-CoV-2 infected mice. The white box indicates the region of magnification. Arrowhead ( N' ) indicates a microglial nodule. Significantly greater Iba1+ area in ( O ) brainstem ( P < 0.001) and ( R ) hippocampus ( P < 0.001) in SARS-CoV-2 infected mice, unpaired t -test. ( S ) SARS-CoV-2 infected mice have significantly lower novel object discrimination in the novel object recognition task for learning and memory than do mock-infected mice ( P < 0.001, unpaired t -test). Discrimination index of 50% indicates novel object preference equal to chance. ( T ) Total object exploration time in the novel object recognition task does not differ between SARS-CoV-2 and mock infected mice, supporting that poor discrimination index in SARS-CoV-2 infected mice is not driven by lack of interest in objects or motility to reach objects. ( U ) SARS-CoV-2 infected mice are significantly slower than mock infected mice to complete the pole descent task for complex motor coordination ( P < 0.0001, unpaired t -test). ( V ) Bouts of spontaneous grooming have longer duration in SARS-CoV-2 infected mice ( P < 0.05, unpaired t -test). ( W ) Fewer individual bouts of spontaneous grooming in SARS-CoV-2 infected mice ( P < 0.05, unpaired t -test).

    Article Snippet: The mouse adapted SARS-CoV-2 MA10 strain was propagated and titred on Vero-E6 cells expressing ACE2 and TMPRSS2 (ATCC, CRL1586).

    Techniques: Permeability, Infection, Staining

    Dysregulation of brain endothelial cell Wnt/β-catenin signalling in SARS-CoV-2 infection. ( A ) Schematic depiction of brain microvascular endothelial cell isolation protocol using gradient centrifugation and positive/negative selection 5 days after respiratory inoculation with SARS-CoV-2. ( B ) Flow cytome­­tric analysis of isolated brain endothelial cells demonstrating uniform population for side-scatter (SSC-H) and forward scatter (FSC-H). ( C ) Flow cytometric analysis showing histogram for CD31 out of all cells in B . Ninety-four per cent of cells were positive for the endothelial cell marker CD31. ( D ) Gene ontology (GO) category analysis of differentially expressed genes in brain endothelial cells of mice with SARS-CoV-2 as compared to mock infection. ( E ) Volcano plot in which differentially expressed genes in KEGG Wnt/β-catenin pathway are depicted in red. Non-differentially expressed KEGG Wnt/β-catenin genes are depicted in pink. Grey dots represent transcripts that are not part of the KEGG Wnt/β-catenin pathway. ( F and G ) Immunostaining for the β-catenin transcriptional target Nkd1 (green) in brain sections of SARS-CoV-2 or mock-infected mice. Brain endothelial cells are visualized with Glut-1 (red). Nuclei are visualized with DAPI (blue). Monochromatic images are provided as . ( H and I ) Quantification of density of Nkd1+ endothelial cells (EC) in the ( H ) hippocampus and ( I ) brainstem of SARS-CoV-2 or mock-infected mice. Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. Unpaired Student’s t -test, ** P < 0.01. FDR = false discovery rate; KEGG = Kyoto Encyclopedia of Genes and Genomes.

    Journal: Brain

    Article Title: Engineered Wnt7a ligands rescue blood–brain barrier and cognitive deficits in a COVID-19 mouse model

    doi: 10.1093/brain/awae031

    Figure Lengend Snippet: Dysregulation of brain endothelial cell Wnt/β-catenin signalling in SARS-CoV-2 infection. ( A ) Schematic depiction of brain microvascular endothelial cell isolation protocol using gradient centrifugation and positive/negative selection 5 days after respiratory inoculation with SARS-CoV-2. ( B ) Flow cytome­­tric analysis of isolated brain endothelial cells demonstrating uniform population for side-scatter (SSC-H) and forward scatter (FSC-H). ( C ) Flow cytometric analysis showing histogram for CD31 out of all cells in B . Ninety-four per cent of cells were positive for the endothelial cell marker CD31. ( D ) Gene ontology (GO) category analysis of differentially expressed genes in brain endothelial cells of mice with SARS-CoV-2 as compared to mock infection. ( E ) Volcano plot in which differentially expressed genes in KEGG Wnt/β-catenin pathway are depicted in red. Non-differentially expressed KEGG Wnt/β-catenin genes are depicted in pink. Grey dots represent transcripts that are not part of the KEGG Wnt/β-catenin pathway. ( F and G ) Immunostaining for the β-catenin transcriptional target Nkd1 (green) in brain sections of SARS-CoV-2 or mock-infected mice. Brain endothelial cells are visualized with Glut-1 (red). Nuclei are visualized with DAPI (blue). Monochromatic images are provided as . ( H and I ) Quantification of density of Nkd1+ endothelial cells (EC) in the ( H ) hippocampus and ( I ) brainstem of SARS-CoV-2 or mock-infected mice. Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. Unpaired Student’s t -test, ** P < 0.01. FDR = false discovery rate; KEGG = Kyoto Encyclopedia of Genes and Genomes.

    Article Snippet: The mouse adapted SARS-CoV-2 MA10 strain was propagated and titred on Vero-E6 cells expressing ACE2 and TMPRSS2 (ATCC, CRL1586).

    Techniques: Infection, Cell Isolation, Gradient Centrifugation, Selection, Isolation, Marker, Immunostaining

    Cerebrovascular-targeted engineered Wnt7a K190A protects against neurobehavioural impairment, neuroinflammation and blood–brain barrier leakage in SARS-CoV-2 infection . Mice were treated with AAV-PHP.eB vector control or AAV-PHP.eB-Wnt7a K190A 18 days prior to inoculation with SARS-CoV-2. Behavioural assessment and tissue collection was conducted 5 days after SARS-CoV-2 inoculation. ( A – F ) Immunostaining for CD3+ T cells (green) in brainstem ( A – C ) and hippocampus ( D – F ) of SARS-CoV-2 infected mice treated with AAV-PHP.eB-Wnt7a K190A or AAV-PHP.eB vector control. Glut-1 (red) was used to visualize endothelial cells. SARS-CoV-2 infected AAV-PHP.eB-Wnt7a K190A treated mice had significantly lower density of CD3+ T cells than did AAV-PHP.eB vector control treated mice in brainstem ( C ) and in hippocampus ( F ). Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. Unpaired Student’s t -test, **** P < 0.0001. ( G – L ) Immunostaining for fibrinogen (red) in brainstem (G–I) and hippocampus ( J – L ) of SARS-CoV-2 infected mice treated with AAV-PHP.eB-Wnt7a K190A or AAV-PHP.eB vector control. Glut1 (green) was used to visualize endothelial cells. SARS-CoV-2 infected AAV-PHP.eB-Wnt7a K190A treated mice had significantly lower perivascular fibrinogen than did AAV-PHP.eB vector control treated mice in brainstem ( I ; unpaired t -test with Welch’s correction for unequal variance, * P < 0.05) and in hippocampus ( L , unpaired t -test, **** P < 0.0001). Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. ( M – R ) Immunostaining for CD68 (green), Iba1 (red), and DAPI (blue) in brainstem ( M and N ) and hippocampus ( P and Q ) of SARS-CoV-2 infected mice treated with AAV-PHP.eB-Wnt7a K190A or AAV-PHP.eB vector control. ( O and R ) Quantification of percent Iba1 + area that is positive for CD68. SARS-CoV-2 infected AAV-PHP.eB-Wnt7a K190A treated mice had significantly lower CD68/Iba1 than did AAV-PHP.eB vector control treated mice in brainstem ( O ; unpaired t -test, ** P < 0.01) and in hippocampus ( R ; unpaired t -test, ** P < 0.01). Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. Monochromatic images are provided in . ( S ) The novel object recognition test for learning and memory was conducted in mice treated with AAV-PHP.eB-Wnt7a K190A or AAV-PHP.eB vector control without or with SARS-CoV-2 infection. There was a Treatment [ F (1,29) = 13.05, P = 0.0011], SARS-CoV-2 [ F (1,29) = 11.75, P = 0.0018], and Treatment × SARS-CoV-2 interaction [ F (1,29) = 8.397, P = 0.0071] for novel object preference. Post hoc analysis revealed that SARS-CoV-2 infection impaired novel object recognition in mice treated with the vector control (**** P < 0.0001) but not in mice treated with AAV-PHP.eB-Wnt7a K190A ( P = 0.7145). Indeed, SARS-CoV-2 infected mice treated with AAV-PHP.eB-Wnt7a K190A performed significantly better than SARS-CoV-2 infected mice treated with the control vector (**** P < 0.0001). ( T ) Total object exploration was not significantly influenced by Treatment [ F (1,29) = 0.009622, P = 0.9225], SARS-CoV-2 [ F (1,29) = 2.679, P = 0.1125], or Treatment × SARS-CoV-2 interaction [ F (1,29) = 0.4577, P = 0.5041], supporting that the novel object recognition task was not confounded by lack of interest or impaired motility. ( U ) Pole descent latency, a measure of complex motor coordination impairment, was significantly influenced by Treatment [ F (1,30) = 7.855, P = 0.0088], SARS-CoV-2 [ F (1,30) = 12.10, P = 0.0016], and Treatment × SARS-CoV-2 interaction [ F (1,30) = 6.410, P = 0.0168]. Post hoc analysis revealed the difference was driven by impairment in the vector-treated SARS-CoV-2 group as compared to the AAV-PHP.eB-Wnt7a K190A treated group (*** P < 0.001) or the uninfected group (*** P < 0.001). ( V ) SARS-CoV-2 infected mice treated with AAV-PHP.eB-Wnt7a K190A fell off the pole significantly fewer times than those treated with AAV-PHP.eB vector control (Fisher’s exact test, * P = 0.0198).

    Journal: Brain

    Article Title: Engineered Wnt7a ligands rescue blood–brain barrier and cognitive deficits in a COVID-19 mouse model

    doi: 10.1093/brain/awae031

    Figure Lengend Snippet: Cerebrovascular-targeted engineered Wnt7a K190A protects against neurobehavioural impairment, neuroinflammation and blood–brain barrier leakage in SARS-CoV-2 infection . Mice were treated with AAV-PHP.eB vector control or AAV-PHP.eB-Wnt7a K190A 18 days prior to inoculation with SARS-CoV-2. Behavioural assessment and tissue collection was conducted 5 days after SARS-CoV-2 inoculation. ( A – F ) Immunostaining for CD3+ T cells (green) in brainstem ( A – C ) and hippocampus ( D – F ) of SARS-CoV-2 infected mice treated with AAV-PHP.eB-Wnt7a K190A or AAV-PHP.eB vector control. Glut-1 (red) was used to visualize endothelial cells. SARS-CoV-2 infected AAV-PHP.eB-Wnt7a K190A treated mice had significantly lower density of CD3+ T cells than did AAV-PHP.eB vector control treated mice in brainstem ( C ) and in hippocampus ( F ). Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. Unpaired Student’s t -test, **** P < 0.0001. ( G – L ) Immunostaining for fibrinogen (red) in brainstem (G–I) and hippocampus ( J – L ) of SARS-CoV-2 infected mice treated with AAV-PHP.eB-Wnt7a K190A or AAV-PHP.eB vector control. Glut1 (green) was used to visualize endothelial cells. SARS-CoV-2 infected AAV-PHP.eB-Wnt7a K190A treated mice had significantly lower perivascular fibrinogen than did AAV-PHP.eB vector control treated mice in brainstem ( I ; unpaired t -test with Welch’s correction for unequal variance, * P < 0.05) and in hippocampus ( L , unpaired t -test, **** P < 0.0001). Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. ( M – R ) Immunostaining for CD68 (green), Iba1 (red), and DAPI (blue) in brainstem ( M and N ) and hippocampus ( P and Q ) of SARS-CoV-2 infected mice treated with AAV-PHP.eB-Wnt7a K190A or AAV-PHP.eB vector control. ( O and R ) Quantification of percent Iba1 + area that is positive for CD68. SARS-CoV-2 infected AAV-PHP.eB-Wnt7a K190A treated mice had significantly lower CD68/Iba1 than did AAV-PHP.eB vector control treated mice in brainstem ( O ; unpaired t -test, ** P < 0.01) and in hippocampus ( R ; unpaired t -test, ** P < 0.01). Each dot represents the average value obtained from two to three tissue sections per mouse. Line indicates group mean. Monochromatic images are provided in . ( S ) The novel object recognition test for learning and memory was conducted in mice treated with AAV-PHP.eB-Wnt7a K190A or AAV-PHP.eB vector control without or with SARS-CoV-2 infection. There was a Treatment [ F (1,29) = 13.05, P = 0.0011], SARS-CoV-2 [ F (1,29) = 11.75, P = 0.0018], and Treatment × SARS-CoV-2 interaction [ F (1,29) = 8.397, P = 0.0071] for novel object preference. Post hoc analysis revealed that SARS-CoV-2 infection impaired novel object recognition in mice treated with the vector control (**** P < 0.0001) but not in mice treated with AAV-PHP.eB-Wnt7a K190A ( P = 0.7145). Indeed, SARS-CoV-2 infected mice treated with AAV-PHP.eB-Wnt7a K190A performed significantly better than SARS-CoV-2 infected mice treated with the control vector (**** P < 0.0001). ( T ) Total object exploration was not significantly influenced by Treatment [ F (1,29) = 0.009622, P = 0.9225], SARS-CoV-2 [ F (1,29) = 2.679, P = 0.1125], or Treatment × SARS-CoV-2 interaction [ F (1,29) = 0.4577, P = 0.5041], supporting that the novel object recognition task was not confounded by lack of interest or impaired motility. ( U ) Pole descent latency, a measure of complex motor coordination impairment, was significantly influenced by Treatment [ F (1,30) = 7.855, P = 0.0088], SARS-CoV-2 [ F (1,30) = 12.10, P = 0.0016], and Treatment × SARS-CoV-2 interaction [ F (1,30) = 6.410, P = 0.0168]. Post hoc analysis revealed the difference was driven by impairment in the vector-treated SARS-CoV-2 group as compared to the AAV-PHP.eB-Wnt7a K190A treated group (*** P < 0.001) or the uninfected group (*** P < 0.001). ( V ) SARS-CoV-2 infected mice treated with AAV-PHP.eB-Wnt7a K190A fell off the pole significantly fewer times than those treated with AAV-PHP.eB vector control (Fisher’s exact test, * P = 0.0198).

    Article Snippet: The mouse adapted SARS-CoV-2 MA10 strain was propagated and titred on Vero-E6 cells expressing ACE2 and TMPRSS2 (ATCC, CRL1586).

    Techniques: Infection, Plasmid Preparation, Immunostaining