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his rbd wt  (Sino Biological)


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    Sino Biological his rbd wt
    SARS-CoV-2-induced CD147 up-regulation causes extended virus infection and pathological injury. a A flow chart for establishing hCD147 mice model of COVID-19. b The virus loads in the lung tissues of hCD147 mice were detected by Taqman-based RT-PCR at 3 and 7 dpi, *** p < 0.001. c , d ,The expression of CD147 was evaluated by western blot in the lung tissues of virus-infected hCD147 mice at 3 and 7 dpi, and the lung tissues of healthy hCD147 mice were used as the control ( c ). The relative expression level of CD147 was analyzed by Image Lab software, *** p < 0.001 ( d ). e The scRNA-seq identified the expression of CD147 in virus-infected ( n = 4,203) and virus-uninfected ( n = 13,354) cells of lung tissues from hCD147 mice. Wilcoxon test, p < 0.0001. f The scRNA-seq identified the expression of CD147 in virus-infected and virus-uninfected AT2 cells of lung tissues from hCD147 mice. g –i Infected, border, and uninfected regions in the lung tissues of virus-infected hCD147 mice were defined using spatial transcriptomics ( g ). The total expression of CD147 was quantified in three defined regions ( h ), and the expression of CD147 in AT2, M2 macrophages, and fibroblasts was determined ( i ). The number of spots used for data analysis in uninfected, border, and infected group were 312, 3,554, and 1,833, respectively. Wilcoxon test, p < 0.05, p < 0.001. j The BEAS-2B cells with SARS-CoV-2 pseudovirus infection in the upper chamber were co-cultured with the BEAS-2B cells in the lower chamber for 48 h, and the expression of CD147 in cells collected from the lower chamber were detected by western blot. k , l The expression of CD147 was determined by western blot in BEAS-2B cells with SARS-CoV-2 pseudovirus infection ( k ) or RBD incubation ( l ). m The relative luciferase signals in pseudovirus-infected BEAS-2B cells were determined by dual-luciferase reporter assays with RBD incubation, *** p < 0.001. n The relative luciferase signals in pseudovirus-infected BEAS-2B cells were determined by dual-luciferase reporter assays, ns, not significant, *** p < 0.001. o The virus loads in authentic virus-infected BEAS-2B cells were determined by Taqman-based RT-PCR, ns, not significant, ** p < 0.01, *** p < 0.001. p A flow chart for establishing a virus-infected hCD147 mouse model by primarily inoculating with RBD protein followed by authentic virus infection. q The expression of CD147 was detected in the lung tissues of hCD147 mice <t>with</t> <t>His-RBD</t> ( n = 4) or His-tag ( n = 4) inoculation and hCD147 mice with His-RBD ( n = 4) or His-tag ( n = 4) inoculation followed by authentic virus infection. r The virus loads in the lung tissues of virus-infected hCD147 mice were determined, * p < 0.05. s , t The infiltration of F4/80+iNOS+ M1 macrophages and Ly6G+ neutrophils in lung tissues of virus-infected hCD147 mice was evaluated by multicolor immunofluorescence staining, Ly6G, yellow; F4/80, green; iNOS, red, scale bar, 100 μm ( s ), and their infiltrated percentages were quantified, ** p < 0.01, *** p < 0.001 ( t ). u The level of cytokines and chemokines in lung tissues of virus-infected hCD147 mice was performed by RT-PCR, * p < 0.05, ** p < 0.01, *** p < 0.001
    His Rbd Wt, supplied by Sino Biological, used in various techniques. Bioz Stars score: 96/100, based on 262 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 96 stars, based on 262 article reviews
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    96/100 stars

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    1) Product Images from "Inducible CD147 up-regulation boosts extended SARS-CoV-2 infection triggering severe COVID-19 independent of ACE2"

    Article Title: Inducible CD147 up-regulation boosts extended SARS-CoV-2 infection triggering severe COVID-19 independent of ACE2

    Journal: Signal Transduction and Targeted Therapy

    doi: 10.1038/s41392-025-02551-x

    SARS-CoV-2-induced CD147 up-regulation causes extended virus infection and pathological injury. a A flow chart for establishing hCD147 mice model of COVID-19. b The virus loads in the lung tissues of hCD147 mice were detected by Taqman-based RT-PCR at 3 and 7 dpi, *** p < 0.001. c , d ,The expression of CD147 was evaluated by western blot in the lung tissues of virus-infected hCD147 mice at 3 and 7 dpi, and the lung tissues of healthy hCD147 mice were used as the control ( c ). The relative expression level of CD147 was analyzed by Image Lab software, *** p < 0.001 ( d ). e The scRNA-seq identified the expression of CD147 in virus-infected ( n = 4,203) and virus-uninfected ( n = 13,354) cells of lung tissues from hCD147 mice. Wilcoxon test, p < 0.0001. f The scRNA-seq identified the expression of CD147 in virus-infected and virus-uninfected AT2 cells of lung tissues from hCD147 mice. g –i Infected, border, and uninfected regions in the lung tissues of virus-infected hCD147 mice were defined using spatial transcriptomics ( g ). The total expression of CD147 was quantified in three defined regions ( h ), and the expression of CD147 in AT2, M2 macrophages, and fibroblasts was determined ( i ). The number of spots used for data analysis in uninfected, border, and infected group were 312, 3,554, and 1,833, respectively. Wilcoxon test, p < 0.05, p < 0.001. j The BEAS-2B cells with SARS-CoV-2 pseudovirus infection in the upper chamber were co-cultured with the BEAS-2B cells in the lower chamber for 48 h, and the expression of CD147 in cells collected from the lower chamber were detected by western blot. k , l The expression of CD147 was determined by western blot in BEAS-2B cells with SARS-CoV-2 pseudovirus infection ( k ) or RBD incubation ( l ). m The relative luciferase signals in pseudovirus-infected BEAS-2B cells were determined by dual-luciferase reporter assays with RBD incubation, *** p < 0.001. n The relative luciferase signals in pseudovirus-infected BEAS-2B cells were determined by dual-luciferase reporter assays, ns, not significant, *** p < 0.001. o The virus loads in authentic virus-infected BEAS-2B cells were determined by Taqman-based RT-PCR, ns, not significant, ** p < 0.01, *** p < 0.001. p A flow chart for establishing a virus-infected hCD147 mouse model by primarily inoculating with RBD protein followed by authentic virus infection. q The expression of CD147 was detected in the lung tissues of hCD147 mice with His-RBD ( n = 4) or His-tag ( n = 4) inoculation and hCD147 mice with His-RBD ( n = 4) or His-tag ( n = 4) inoculation followed by authentic virus infection. r The virus loads in the lung tissues of virus-infected hCD147 mice were determined, * p < 0.05. s , t The infiltration of F4/80+iNOS+ M1 macrophages and Ly6G+ neutrophils in lung tissues of virus-infected hCD147 mice was evaluated by multicolor immunofluorescence staining, Ly6G, yellow; F4/80, green; iNOS, red, scale bar, 100 μm ( s ), and their infiltrated percentages were quantified, ** p < 0.01, *** p < 0.001 ( t ). u The level of cytokines and chemokines in lung tissues of virus-infected hCD147 mice was performed by RT-PCR, * p < 0.05, ** p < 0.01, *** p < 0.001
    Figure Legend Snippet: SARS-CoV-2-induced CD147 up-regulation causes extended virus infection and pathological injury. a A flow chart for establishing hCD147 mice model of COVID-19. b The virus loads in the lung tissues of hCD147 mice were detected by Taqman-based RT-PCR at 3 and 7 dpi, *** p < 0.001. c , d ,The expression of CD147 was evaluated by western blot in the lung tissues of virus-infected hCD147 mice at 3 and 7 dpi, and the lung tissues of healthy hCD147 mice were used as the control ( c ). The relative expression level of CD147 was analyzed by Image Lab software, *** p < 0.001 ( d ). e The scRNA-seq identified the expression of CD147 in virus-infected ( n = 4,203) and virus-uninfected ( n = 13,354) cells of lung tissues from hCD147 mice. Wilcoxon test, p < 0.0001. f The scRNA-seq identified the expression of CD147 in virus-infected and virus-uninfected AT2 cells of lung tissues from hCD147 mice. g –i Infected, border, and uninfected regions in the lung tissues of virus-infected hCD147 mice were defined using spatial transcriptomics ( g ). The total expression of CD147 was quantified in three defined regions ( h ), and the expression of CD147 in AT2, M2 macrophages, and fibroblasts was determined ( i ). The number of spots used for data analysis in uninfected, border, and infected group were 312, 3,554, and 1,833, respectively. Wilcoxon test, p < 0.05, p < 0.001. j The BEAS-2B cells with SARS-CoV-2 pseudovirus infection in the upper chamber were co-cultured with the BEAS-2B cells in the lower chamber for 48 h, and the expression of CD147 in cells collected from the lower chamber were detected by western blot. k , l The expression of CD147 was determined by western blot in BEAS-2B cells with SARS-CoV-2 pseudovirus infection ( k ) or RBD incubation ( l ). m The relative luciferase signals in pseudovirus-infected BEAS-2B cells were determined by dual-luciferase reporter assays with RBD incubation, *** p < 0.001. n The relative luciferase signals in pseudovirus-infected BEAS-2B cells were determined by dual-luciferase reporter assays, ns, not significant, *** p < 0.001. o The virus loads in authentic virus-infected BEAS-2B cells were determined by Taqman-based RT-PCR, ns, not significant, ** p < 0.01, *** p < 0.001. p A flow chart for establishing a virus-infected hCD147 mouse model by primarily inoculating with RBD protein followed by authentic virus infection. q The expression of CD147 was detected in the lung tissues of hCD147 mice with His-RBD ( n = 4) or His-tag ( n = 4) inoculation and hCD147 mice with His-RBD ( n = 4) or His-tag ( n = 4) inoculation followed by authentic virus infection. r The virus loads in the lung tissues of virus-infected hCD147 mice were determined, * p < 0.05. s , t The infiltration of F4/80+iNOS+ M1 macrophages and Ly6G+ neutrophils in lung tissues of virus-infected hCD147 mice was evaluated by multicolor immunofluorescence staining, Ly6G, yellow; F4/80, green; iNOS, red, scale bar, 100 μm ( s ), and their infiltrated percentages were quantified, ** p < 0.01, *** p < 0.001 ( t ). u The level of cytokines and chemokines in lung tissues of virus-infected hCD147 mice was performed by RT-PCR, * p < 0.05, ** p < 0.01, *** p < 0.001

    Techniques Used: Virus, Infection, Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot, Control, Software, Cell Culture, Incubation, Luciferase, Multicolor Immunofluorescence Staining



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    Sangon Biotech fitc-tagged wt-rbd protein
    SARS-CoV-2-induced CD147 up-regulation causes extended virus infection and pathological injury. a A flow chart for establishing hCD147 mice model of COVID-19. b The virus loads in the lung tissues of hCD147 mice were detected by Taqman-based RT-PCR at 3 and 7 dpi, *** p < 0.001. c , d ,The expression of CD147 was evaluated by western blot in the lung tissues of virus-infected hCD147 mice at 3 and 7 dpi, and the lung tissues of healthy hCD147 mice were used as the control ( c ). The relative expression level of CD147 was analyzed by Image Lab software, *** p < 0.001 ( d ). e The scRNA-seq identified the expression of CD147 in virus-infected ( n = 4,203) and virus-uninfected ( n = 13,354) cells of lung tissues from hCD147 mice. Wilcoxon test, p < 0.0001. f The scRNA-seq identified the expression of CD147 in virus-infected and virus-uninfected AT2 cells of lung tissues from hCD147 mice. g –i Infected, border, and uninfected regions in the lung tissues of virus-infected hCD147 mice were defined using spatial transcriptomics ( g ). The total expression of CD147 was quantified in three defined regions ( h ), and the expression of CD147 in AT2, M2 macrophages, and fibroblasts was determined ( i ). The number of spots used for data analysis in uninfected, border, and infected group were 312, 3,554, and 1,833, respectively. Wilcoxon test, p < 0.05, p < 0.001. j The BEAS-2B cells with SARS-CoV-2 pseudovirus infection in the upper chamber were co-cultured with the BEAS-2B cells in the lower chamber for 48 h, and the expression of CD147 in cells collected from the lower chamber were detected by western blot. k , l The expression of CD147 was determined by western blot in BEAS-2B cells with SARS-CoV-2 pseudovirus infection ( k ) or RBD incubation ( l ). m The relative luciferase signals in pseudovirus-infected BEAS-2B cells were determined by dual-luciferase reporter assays with RBD incubation, *** p < 0.001. n The relative luciferase signals in pseudovirus-infected BEAS-2B cells were determined by dual-luciferase reporter assays, ns, not significant, *** p < 0.001. o The virus loads in authentic virus-infected BEAS-2B cells were determined by Taqman-based RT-PCR, ns, not significant, ** p < 0.01, *** p < 0.001. p A flow chart for establishing a virus-infected hCD147 mouse model by primarily inoculating with RBD protein followed by authentic virus infection. q The expression of CD147 was detected in the lung tissues of hCD147 mice <t>with</t> <t>His-RBD</t> ( n = 4) or His-tag ( n = 4) inoculation and hCD147 mice with His-RBD ( n = 4) or His-tag ( n = 4) inoculation followed by authentic virus infection. r The virus loads in the lung tissues of virus-infected hCD147 mice were determined, * p < 0.05. s , t The infiltration of F4/80+iNOS+ M1 macrophages and Ly6G+ neutrophils in lung tissues of virus-infected hCD147 mice was evaluated by multicolor immunofluorescence staining, Ly6G, yellow; F4/80, green; iNOS, red, scale bar, 100 μm ( s ), and their infiltrated percentages were quantified, ** p < 0.01, *** p < 0.001 ( t ). u The level of cytokines and chemokines in lung tissues of virus-infected hCD147 mice was performed by RT-PCR, * p < 0.05, ** p < 0.01, *** p < 0.001
    Fitc Tagged Wt Rbd Protein, supplied by Sangon Biotech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    wt rbd  (Sino Biological)
    94
    Sino Biological wt rbd
    Memory B cell responses after an extended period post-BA.5 infection. (A) Timeline of vaccination, infection, and blood draws for 3 human cohorts, including short-term post-BA.5/BF.7 BTI sampling at 1 month (BA.5/BF.7 BTI (1 m)), prolonged post-BA.5/BF.7 BTI sampling at 7 months (BA.5/BF.7 BTI (7 m)), and prolonged post-BA.5/BF.7 infection sampling at 8 months (BA.5/BF.7 inf. (8 m)). (B) Flow cytometry analysis of pooled memory B cells from three cohorts including BA.5 (1 m), BA.5/BF.7 BTI (7 m), and BA.5/BF.7 inf. (8 m). BA.5 <t>RBD</t> <t>APC</t> and PE double-positive memory B cells (left panel) were analyzed for cross-reactivity with the wide-type (WT) RBD (right panel).
    Wt Rbd, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rbds  (Addgene inc)
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    Fig. 1 | Preparation of multiviral Quartet Nanocages. a, Plug-and-Display vaccine assembly of mosaic and Quartet Nanocages. Genetic fusion of SpyCatcher003 (dark blue) with mi3 (purple) allows efficient multimerization of single or Quartet <t>RBDs</t> linked <t>to</t> <t>SpyTag</t> (cyan) through spontaneous isopeptide bond formation (marked in red). Only some antigens are shown in the schematic for clarity. b, Phylogenetic tree of sarbecoviruses used in this study, based on RBD sequence. c, Genetic organization of the multiviral Quartet-SpyTag, indicating the viral origin of RBDs, N-linked glycosylation sites and tag location. d, Analysis of Quartet-SpyTag with SDS–PAGE/Coomassie staining, with or without PNGase F deglycosylation. A representative gel from two independent experiments. Molecular weight markers are in kDa. e, Coupling of RBD Quartet to SpyCatcher003-mi3 Nanocage at different molar Nanocage:antigen ratios, analysed by SDS–PAGE/Coomassie. A representative gel from two independent experiments. Molecular weight markers are in kDa.
    Rbds, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    wt sars cov 2 rbd  (Addgene inc)
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    Fig. 1 | Preparation of multiviral Quartet Nanocages. a, Plug-and-Display vaccine assembly of mosaic and Quartet Nanocages. Genetic fusion of SpyCatcher003 (dark blue) with mi3 (purple) allows efficient multimerization of single or Quartet <t>RBDs</t> linked <t>to</t> <t>SpyTag</t> (cyan) through spontaneous isopeptide bond formation (marked in red). Only some antigens are shown in the schematic for clarity. b, Phylogenetic tree of sarbecoviruses used in this study, based on RBD sequence. c, Genetic organization of the multiviral Quartet-SpyTag, indicating the viral origin of RBDs, N-linked glycosylation sites and tag location. d, Analysis of Quartet-SpyTag with SDS–PAGE/Coomassie staining, with or without PNGase F deglycosylation. A representative gel from two independent experiments. Molecular weight markers are in kDa. e, Coupling of RBD Quartet to SpyCatcher003-mi3 Nanocage at different molar Nanocage:antigen ratios, analysed by SDS–PAGE/Coomassie. A representative gel from two independent experiments. Molecular weight markers are in kDa.
    Wt Sars Cov 2 Rbd, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    SARS-CoV-2-induced CD147 up-regulation causes extended virus infection and pathological injury. a A flow chart for establishing hCD147 mice model of COVID-19. b The virus loads in the lung tissues of hCD147 mice were detected by Taqman-based RT-PCR at 3 and 7 dpi, *** p < 0.001. c , d ,The expression of CD147 was evaluated by western blot in the lung tissues of virus-infected hCD147 mice at 3 and 7 dpi, and the lung tissues of healthy hCD147 mice were used as the control ( c ). The relative expression level of CD147 was analyzed by Image Lab software, *** p < 0.001 ( d ). e The scRNA-seq identified the expression of CD147 in virus-infected ( n = 4,203) and virus-uninfected ( n = 13,354) cells of lung tissues from hCD147 mice. Wilcoxon test, p < 0.0001. f The scRNA-seq identified the expression of CD147 in virus-infected and virus-uninfected AT2 cells of lung tissues from hCD147 mice. g –i Infected, border, and uninfected regions in the lung tissues of virus-infected hCD147 mice were defined using spatial transcriptomics ( g ). The total expression of CD147 was quantified in three defined regions ( h ), and the expression of CD147 in AT2, M2 macrophages, and fibroblasts was determined ( i ). The number of spots used for data analysis in uninfected, border, and infected group were 312, 3,554, and 1,833, respectively. Wilcoxon test, p < 0.05, p < 0.001. j The BEAS-2B cells with SARS-CoV-2 pseudovirus infection in the upper chamber were co-cultured with the BEAS-2B cells in the lower chamber for 48 h, and the expression of CD147 in cells collected from the lower chamber were detected by western blot. k , l The expression of CD147 was determined by western blot in BEAS-2B cells with SARS-CoV-2 pseudovirus infection ( k ) or RBD incubation ( l ). m The relative luciferase signals in pseudovirus-infected BEAS-2B cells were determined by dual-luciferase reporter assays with RBD incubation, *** p < 0.001. n The relative luciferase signals in pseudovirus-infected BEAS-2B cells were determined by dual-luciferase reporter assays, ns, not significant, *** p < 0.001. o The virus loads in authentic virus-infected BEAS-2B cells were determined by Taqman-based RT-PCR, ns, not significant, ** p < 0.01, *** p < 0.001. p A flow chart for establishing a virus-infected hCD147 mouse model by primarily inoculating with RBD protein followed by authentic virus infection. q The expression of CD147 was detected in the lung tissues of hCD147 mice with His-RBD ( n = 4) or His-tag ( n = 4) inoculation and hCD147 mice with His-RBD ( n = 4) or His-tag ( n = 4) inoculation followed by authentic virus infection. r The virus loads in the lung tissues of virus-infected hCD147 mice were determined, * p < 0.05. s , t The infiltration of F4/80+iNOS+ M1 macrophages and Ly6G+ neutrophils in lung tissues of virus-infected hCD147 mice was evaluated by multicolor immunofluorescence staining, Ly6G, yellow; F4/80, green; iNOS, red, scale bar, 100 μm ( s ), and their infiltrated percentages were quantified, ** p < 0.01, *** p < 0.001 ( t ). u The level of cytokines and chemokines in lung tissues of virus-infected hCD147 mice was performed by RT-PCR, * p < 0.05, ** p < 0.01, *** p < 0.001

    Journal: Signal Transduction and Targeted Therapy

    Article Title: Inducible CD147 up-regulation boosts extended SARS-CoV-2 infection triggering severe COVID-19 independent of ACE2

    doi: 10.1038/s41392-025-02551-x

    Figure Lengend Snippet: SARS-CoV-2-induced CD147 up-regulation causes extended virus infection and pathological injury. a A flow chart for establishing hCD147 mice model of COVID-19. b The virus loads in the lung tissues of hCD147 mice were detected by Taqman-based RT-PCR at 3 and 7 dpi, *** p < 0.001. c , d ,The expression of CD147 was evaluated by western blot in the lung tissues of virus-infected hCD147 mice at 3 and 7 dpi, and the lung tissues of healthy hCD147 mice were used as the control ( c ). The relative expression level of CD147 was analyzed by Image Lab software, *** p < 0.001 ( d ). e The scRNA-seq identified the expression of CD147 in virus-infected ( n = 4,203) and virus-uninfected ( n = 13,354) cells of lung tissues from hCD147 mice. Wilcoxon test, p < 0.0001. f The scRNA-seq identified the expression of CD147 in virus-infected and virus-uninfected AT2 cells of lung tissues from hCD147 mice. g –i Infected, border, and uninfected regions in the lung tissues of virus-infected hCD147 mice were defined using spatial transcriptomics ( g ). The total expression of CD147 was quantified in three defined regions ( h ), and the expression of CD147 in AT2, M2 macrophages, and fibroblasts was determined ( i ). The number of spots used for data analysis in uninfected, border, and infected group were 312, 3,554, and 1,833, respectively. Wilcoxon test, p < 0.05, p < 0.001. j The BEAS-2B cells with SARS-CoV-2 pseudovirus infection in the upper chamber were co-cultured with the BEAS-2B cells in the lower chamber for 48 h, and the expression of CD147 in cells collected from the lower chamber were detected by western blot. k , l The expression of CD147 was determined by western blot in BEAS-2B cells with SARS-CoV-2 pseudovirus infection ( k ) or RBD incubation ( l ). m The relative luciferase signals in pseudovirus-infected BEAS-2B cells were determined by dual-luciferase reporter assays with RBD incubation, *** p < 0.001. n The relative luciferase signals in pseudovirus-infected BEAS-2B cells were determined by dual-luciferase reporter assays, ns, not significant, *** p < 0.001. o The virus loads in authentic virus-infected BEAS-2B cells were determined by Taqman-based RT-PCR, ns, not significant, ** p < 0.01, *** p < 0.001. p A flow chart for establishing a virus-infected hCD147 mouse model by primarily inoculating with RBD protein followed by authentic virus infection. q The expression of CD147 was detected in the lung tissues of hCD147 mice with His-RBD ( n = 4) or His-tag ( n = 4) inoculation and hCD147 mice with His-RBD ( n = 4) or His-tag ( n = 4) inoculation followed by authentic virus infection. r The virus loads in the lung tissues of virus-infected hCD147 mice were determined, * p < 0.05. s , t The infiltration of F4/80+iNOS+ M1 macrophages and Ly6G+ neutrophils in lung tissues of virus-infected hCD147 mice was evaluated by multicolor immunofluorescence staining, Ly6G, yellow; F4/80, green; iNOS, red, scale bar, 100 μm ( s ), and their infiltrated percentages were quantified, ** p < 0.01, *** p < 0.001 ( t ). u The level of cytokines and chemokines in lung tissues of virus-infected hCD147 mice was performed by RT-PCR, * p < 0.05, ** p < 0.01, *** p < 0.001

    Article Snippet: The mobile phases contained the mutants of CD147 (His-CD147-R54A, His-CD147-E84A, His-CD147-E92A, His-CD147-Q100A, His-CD147-S112A, our laboratory), the mutants of RBD (His-RBD-G413A, His-RBD-K417A, His-RBD-K424A, His-RBD-G447A, His-RBD-Y489A, Sino Biological), His-RBD (WT) (40592-V08H, Sino Biological), His-RBD (Beta) (40592-V08H85, Sino Biological), His-RBD (Gamma) (40592-V08H86, Sino Biological), His-RBD (JN.1) (40592-V08H155, Sino Biological), rhesus macaque CD147 (Sino Biological), human ACE2 (Sino Biological), and rhesus macaque ACE2 (Sino Biological).

    Techniques: Virus, Infection, Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot, Control, Software, Cell Culture, Incubation, Luciferase, Multicolor Immunofluorescence Staining

    Memory B cell responses after an extended period post-BA.5 infection. (A) Timeline of vaccination, infection, and blood draws for 3 human cohorts, including short-term post-BA.5/BF.7 BTI sampling at 1 month (BA.5/BF.7 BTI (1 m)), prolonged post-BA.5/BF.7 BTI sampling at 7 months (BA.5/BF.7 BTI (7 m)), and prolonged post-BA.5/BF.7 infection sampling at 8 months (BA.5/BF.7 inf. (8 m)). (B) Flow cytometry analysis of pooled memory B cells from three cohorts including BA.5 (1 m), BA.5/BF.7 BTI (7 m), and BA.5/BF.7 inf. (8 m). BA.5 RBD APC and PE double-positive memory B cells (left panel) were analyzed for cross-reactivity with the wide-type (WT) RBD (right panel).

    Journal: Emerging Microbes & Infections

    Article Title: Prolonged Omicron-specific B cell maturation alleviates immune imprinting induced by SARS-CoV-2 inactivated vaccine

    doi: 10.1080/22221751.2024.2412623

    Figure Lengend Snippet: Memory B cell responses after an extended period post-BA.5 infection. (A) Timeline of vaccination, infection, and blood draws for 3 human cohorts, including short-term post-BA.5/BF.7 BTI sampling at 1 month (BA.5/BF.7 BTI (1 m)), prolonged post-BA.5/BF.7 BTI sampling at 7 months (BA.5/BF.7 BTI (7 m)), and prolonged post-BA.5/BF.7 infection sampling at 8 months (BA.5/BF.7 inf. (8 m)). (B) Flow cytometry analysis of pooled memory B cells from three cohorts including BA.5 (1 m), BA.5/BF.7 BTI (7 m), and BA.5/BF.7 inf. (8 m). BA.5 RBD APC and PE double-positive memory B cells (left panel) were analyzed for cross-reactivity with the wide-type (WT) RBD (right panel).

    Article Snippet: Additionally, 0.013 μg of BA.5 RBD (Sino Biological, 40592-V49H9-B) tagged with PE-streptavidin (Biolegend, 405204) and APC-streptavidin (Biolegend, 405207), and 0.013 μg of WT RBD (Sino Biological, 40592-V27H-B) tagged with BV605 streptavidin (Biolegend, 405229) was added.

    Techniques: Infection, Sampling, Flow Cytometry

    Fig. 1 | Preparation of multiviral Quartet Nanocages. a, Plug-and-Display vaccine assembly of mosaic and Quartet Nanocages. Genetic fusion of SpyCatcher003 (dark blue) with mi3 (purple) allows efficient multimerization of single or Quartet RBDs linked to SpyTag (cyan) through spontaneous isopeptide bond formation (marked in red). Only some antigens are shown in the schematic for clarity. b, Phylogenetic tree of sarbecoviruses used in this study, based on RBD sequence. c, Genetic organization of the multiviral Quartet-SpyTag, indicating the viral origin of RBDs, N-linked glycosylation sites and tag location. d, Analysis of Quartet-SpyTag with SDS–PAGE/Coomassie staining, with or without PNGase F deglycosylation. A representative gel from two independent experiments. Molecular weight markers are in kDa. e, Coupling of RBD Quartet to SpyCatcher003-mi3 Nanocage at different molar Nanocage:antigen ratios, analysed by SDS–PAGE/Coomassie. A representative gel from two independent experiments. Molecular weight markers are in kDa.

    Journal: Nature nanotechnology

    Article Title: Proactive vaccination using multiviral Quartet Nanocages to elicit broad anti-coronavirus responses.

    doi: 10.1038/s41565-024-01655-9

    Figure Lengend Snippet: Fig. 1 | Preparation of multiviral Quartet Nanocages. a, Plug-and-Display vaccine assembly of mosaic and Quartet Nanocages. Genetic fusion of SpyCatcher003 (dark blue) with mi3 (purple) allows efficient multimerization of single or Quartet RBDs linked to SpyTag (cyan) through spontaneous isopeptide bond formation (marked in red). Only some antigens are shown in the schematic for clarity. b, Phylogenetic tree of sarbecoviruses used in this study, based on RBD sequence. c, Genetic organization of the multiviral Quartet-SpyTag, indicating the viral origin of RBDs, N-linked glycosylation sites and tag location. d, Analysis of Quartet-SpyTag with SDS–PAGE/Coomassie staining, with or without PNGase F deglycosylation. A representative gel from two independent experiments. Molecular weight markers are in kDa. e, Coupling of RBD Quartet to SpyCatcher003-mi3 Nanocage at different molar Nanocage:antigen ratios, analysed by SDS–PAGE/Coomassie. A representative gel from two independent experiments. Molecular weight markers are in kDa.

    Article Snippet: For subsequent figures, pcDNA3.1-SpyTag-Quartet was cloned with a SpyTag after the signal sequence and then the same order of RBDs (SpyTag-SHC014-Rs4081-RaTG13-SARS2) (Supplementary Fig. 8; GenBank PP136031, Addgene Plasmid ID 214727). pcDNA3.1-Quartet [SARS1] was cloned with SpyTag after the signal sequence, with SARS1 in the position of SARS2 (SpyTag-SHC014-Rs4081-RaTG13-SARS1) (Supplementary Fig. 8; GenBank PP136034, Addgene plasmid ID 214729). pcDNA3.1-Alternate Quartet was cloned with SpyTag after the signal sequence, followed by pang17 RBD, RmYN02 RBD, Rf1 RBD and WIV1 RBD (Supplementary Fig. 8; GenBank PP136032, Addgene plasmid ID 214728). pcDNA3.1-SpyTag-Quartet_NoLinker was cloned with the same order of RBDs as SpyTag-Quartet (SpyTag-SHC014-Rs4081RaTG13-SARS2) but did not have any Gly-Ser linker between RBDs (Supplementary Fig. 8; GenBank PP136036, Addgene plasmid ID 214731). pcDNA3.1-Kraken Quartet was identical to SpyTag-Quartet with the SARS2 XBB.1.5 RBD in place of SARS2 Wuhan RBD (Supplementary Fig. 8; GenBank PP136035, Addgene plasmid ID 214730).

    Techniques: Sequencing, Glycoproteomics, SDS Page, Staining, Molecular Weight

    Fig. 2 | Broad immune response from immunization with Quartet Nanocages. a, Schematic of antigens for this set of immunizations, comparing uncoupled proteins or proteins coupled to the SpyCatcher003-mi3 nanocage. b, Procedure for immunization and sampling. c, ELISA for post-boost serum IgG binding to different sarbecovirus RBDs is shown as the area under the curve (AUC) of a serial dilution. Sera are from mice immunized with uncoupled SARS2 Wuhan RBD (orange), uncoupled Quartet-SpyTag (yellow), SARS2 Wuhan RBD coupled to SpyCatcher003-mi3 (green) or Quartet-SpyTag coupled to SpyCatcher003-mi3 (blue). Solid rectangles under samples indicate ELISA against a component of that vaccine (matched). Striped rectangles indicate ELISA against an antigen absent in that vaccine (mismatched). Each dot represents one animal. The mean is denoted by a bar, shown ±1 s.d.; n = 6. Significance was calculated with an ANOVA test using Tukey’s post hoc test. *P < 0.05, **P < 0.01, ***P < 0.001; other comparisons were non-significant.

    Journal: Nature nanotechnology

    Article Title: Proactive vaccination using multiviral Quartet Nanocages to elicit broad anti-coronavirus responses.

    doi: 10.1038/s41565-024-01655-9

    Figure Lengend Snippet: Fig. 2 | Broad immune response from immunization with Quartet Nanocages. a, Schematic of antigens for this set of immunizations, comparing uncoupled proteins or proteins coupled to the SpyCatcher003-mi3 nanocage. b, Procedure for immunization and sampling. c, ELISA for post-boost serum IgG binding to different sarbecovirus RBDs is shown as the area under the curve (AUC) of a serial dilution. Sera are from mice immunized with uncoupled SARS2 Wuhan RBD (orange), uncoupled Quartet-SpyTag (yellow), SARS2 Wuhan RBD coupled to SpyCatcher003-mi3 (green) or Quartet-SpyTag coupled to SpyCatcher003-mi3 (blue). Solid rectangles under samples indicate ELISA against a component of that vaccine (matched). Striped rectangles indicate ELISA against an antigen absent in that vaccine (mismatched). Each dot represents one animal. The mean is denoted by a bar, shown ±1 s.d.; n = 6. Significance was calculated with an ANOVA test using Tukey’s post hoc test. *P < 0.05, **P < 0.01, ***P < 0.001; other comparisons were non-significant.

    Article Snippet: For subsequent figures, pcDNA3.1-SpyTag-Quartet was cloned with a SpyTag after the signal sequence and then the same order of RBDs (SpyTag-SHC014-Rs4081-RaTG13-SARS2) (Supplementary Fig. 8; GenBank PP136031, Addgene Plasmid ID 214727). pcDNA3.1-Quartet [SARS1] was cloned with SpyTag after the signal sequence, with SARS1 in the position of SARS2 (SpyTag-SHC014-Rs4081-RaTG13-SARS1) (Supplementary Fig. 8; GenBank PP136034, Addgene plasmid ID 214729). pcDNA3.1-Alternate Quartet was cloned with SpyTag after the signal sequence, followed by pang17 RBD, RmYN02 RBD, Rf1 RBD and WIV1 RBD (Supplementary Fig. 8; GenBank PP136032, Addgene plasmid ID 214728). pcDNA3.1-SpyTag-Quartet_NoLinker was cloned with the same order of RBDs as SpyTag-Quartet (SpyTag-SHC014-Rs4081RaTG13-SARS2) but did not have any Gly-Ser linker between RBDs (Supplementary Fig. 8; GenBank PP136036, Addgene plasmid ID 214731). pcDNA3.1-Kraken Quartet was identical to SpyTag-Quartet with the SARS2 XBB.1.5 RBD in place of SARS2 Wuhan RBD (Supplementary Fig. 8; GenBank PP136035, Addgene plasmid ID 214730).

    Techniques: Sampling, Enzyme-linked Immunosorbent Assay, Binding Assay, Serial Dilution

    Fig. 5 | Quartet immunization induces broad antibodies even after a preprimed SARS2 response. a, Summary of timeline and antigens for this set of immunizations. b, ELISA for serum IgG to SARS2 Wuhan RBD presented as the AUC of a serial dilution. All mice were primed with Wuhan SARS2 Spike, before boosting with Wuhan SARS2 Spike protein (light green), Homotypic Nanocage (pink), Mosaic-8 (dark blue), SpyTag-Quartet Nanocage (red), Dual Quartet Nanocage (orange), Quartet Nanocage with SARS1 RBD replacing SARS2 (purple) or Dual Quartet Nanocage with SARS1 RBD replacing SARS2 (cyan). Solid rectangles under samples indicate ELISA against a component of that vaccine (matched). Striped rectangles indicate ELISA against an antigen absent in that vaccine (mismatched). Each dot represents one animal. The mean is denoted by a bar ±1 s.d.; n = 6. c, ELISA for serum IgG to other sarbecovirus RBDs, as for b, with each dot representing one animal and the mean being denoted by a bar ±1 s.d.; n = 6. Significance was calculated with an ANOVA test using Tukey’s post hoc test. *P < 0.05, **P < 0.01, ***P < 0.001; other comparisons were non-significant.

    Journal: Nature nanotechnology

    Article Title: Proactive vaccination using multiviral Quartet Nanocages to elicit broad anti-coronavirus responses.

    doi: 10.1038/s41565-024-01655-9

    Figure Lengend Snippet: Fig. 5 | Quartet immunization induces broad antibodies even after a preprimed SARS2 response. a, Summary of timeline and antigens for this set of immunizations. b, ELISA for serum IgG to SARS2 Wuhan RBD presented as the AUC of a serial dilution. All mice were primed with Wuhan SARS2 Spike, before boosting with Wuhan SARS2 Spike protein (light green), Homotypic Nanocage (pink), Mosaic-8 (dark blue), SpyTag-Quartet Nanocage (red), Dual Quartet Nanocage (orange), Quartet Nanocage with SARS1 RBD replacing SARS2 (purple) or Dual Quartet Nanocage with SARS1 RBD replacing SARS2 (cyan). Solid rectangles under samples indicate ELISA against a component of that vaccine (matched). Striped rectangles indicate ELISA against an antigen absent in that vaccine (mismatched). Each dot represents one animal. The mean is denoted by a bar ±1 s.d.; n = 6. c, ELISA for serum IgG to other sarbecovirus RBDs, as for b, with each dot representing one animal and the mean being denoted by a bar ±1 s.d.; n = 6. Significance was calculated with an ANOVA test using Tukey’s post hoc test. *P < 0.05, **P < 0.01, ***P < 0.001; other comparisons were non-significant.

    Article Snippet: For subsequent figures, pcDNA3.1-SpyTag-Quartet was cloned with a SpyTag after the signal sequence and then the same order of RBDs (SpyTag-SHC014-Rs4081-RaTG13-SARS2) (Supplementary Fig. 8; GenBank PP136031, Addgene Plasmid ID 214727). pcDNA3.1-Quartet [SARS1] was cloned with SpyTag after the signal sequence, with SARS1 in the position of SARS2 (SpyTag-SHC014-Rs4081-RaTG13-SARS1) (Supplementary Fig. 8; GenBank PP136034, Addgene plasmid ID 214729). pcDNA3.1-Alternate Quartet was cloned with SpyTag after the signal sequence, followed by pang17 RBD, RmYN02 RBD, Rf1 RBD and WIV1 RBD (Supplementary Fig. 8; GenBank PP136032, Addgene plasmid ID 214728). pcDNA3.1-SpyTag-Quartet_NoLinker was cloned with the same order of RBDs as SpyTag-Quartet (SpyTag-SHC014-Rs4081RaTG13-SARS2) but did not have any Gly-Ser linker between RBDs (Supplementary Fig. 8; GenBank PP136036, Addgene plasmid ID 214731). pcDNA3.1-Kraken Quartet was identical to SpyTag-Quartet with the SARS2 XBB.1.5 RBD in place of SARS2 Wuhan RBD (Supplementary Fig. 8; GenBank PP136035, Addgene plasmid ID 214730).

    Techniques: Enzyme-linked Immunosorbent Assay, Serial Dilution