his rbd protein  (Sino Biological)

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: His-RBD protein (500 ng/mL, 40592-V08H, Sino Biological) and 6×His-tag (500 ng/mL, QYAOBIO) were added in the medium for 24 h. Then, the cell medium was discarded and the authentic SARS-CoV-2 virus (Beta, 100 TCID 50 ) was added into each well for 2 h. Subsequently, the cell supernatant was replaced with 2% FBS medium and the cells were cultured at 37 °C for 48 h. Finally, the samples were collected and the virus loads were detected by Taqman-based real-time PCR.

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

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

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: Cryo-EM structure demonstrates CD147-spike interaction. a Cryo-EM map of the CD147-spike complex. The RBD up protomer, green; the two RBD down protomers, sandy brown and blue; CD147, magenta. b The overall structure of the CD147-spike complex shown as cartoons in side (left) and top (right) views. c The overlaid structures of CD147-spike and spike-closed are shown in surface representation. CD147 is depicted in magenta, with the spike protomer in the up and down conformations shown in green and cyan, respectively. The contact patch recognized by CD147 is highlighted in yellow on the down RBD. d The structure and key residues within the binding interface of the CD147-spike complex. The key residues forming hydrogen bonds are presented by stick models, with the hydrogen-bonding interactions labeled in yellow lines. e The binding ability of spike RBD with wildtype CD147 or its mutants (CD147-R54A, E84A, E92A, Q100A, and S112A) and CD147 with wildtype spike RBD or its mutants (G413A, K417A, K424A, G447A, and Y489A) was determined by SPR assay. f The binding ability of CD147 with wildtype spike RBD or its mutants (spike RBD-Beta, Gamma, and JN.1) was determined by SPR assay. g The virus loads and relative luciferase signals in CD147/ACE2 double knockout Vero E6 cells transfected with either wild type CD147 or mutated CD147 were determined by Taqman-based RT-PCR and dual-luciferase reporter assays, respectively, *** p < 0.001. h The overlaid structures of the CD147-spike complex, colored as in (a), and the CD147-Meplazumab Fab complex (PDB ID: 5X0T), colored in gray. The up RBD and Meplazumab Fab are depicted as surfaces, with the binding epitopes on CD147 (residues 61-75, colored in red) recognized by Meplazumab. The steric hindrance between Meplazumab and the spike bound to CD147 is indicated with orange arrows. i The competitive inhibitory role of MPZ for the binding of CD147 and SARS-CoV-2 spike (RBD) protein was determined by ELISA assay

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: Cryo-EM Sample Prep, Binding Assay, Labeling, SPR Assay, Virus, Luciferase, Double Knockout, Transfection, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

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

Journal: npj Viruses

Article Title: K5 polysaccharides inhibit SARS-CoV-2 infection by preventing spike-proteolytic priming

doi: 10.1038/s44298-025-00163-4

Figure Lengend Snippet: a Representative images of syncytia formation assay in VeroE6 cells upon treatment (350 µM) with K5 compounds. Scale bar: 50 µm. b Number of nuclei involved in syncytia formation is higher in Wuhan-Hu-1 spike-positive cells than in Omicron BA.1 spike-positive cells. c Effect of K5 compounds on syncytia formation induced by Wuhan-Hu-1 spike. d Effect of K5 compounds on syncytia formation induced by Omicron BA.1 spike. Only spike-positive cells were quantified. Data of four (Wuhan-Hu-1) and three (Omicron BA.1) independent experiments. Values are mean ± sd. P values determined by Welch’s t-test: * P < 0.05; ** P < 0.005.

Article Snippet: Reagents and materials were used as received, unless otherwise mentioned, and were purchased from the following: Human recombinant SARS-CoV-2 Wuhan-Hu-1 spike His-Tag protein and RBD from Sino Biological (#40592-V08B); ACE2 from Acrobiosystem (#AC2-H52H8); Bovine Serum Albumin (BSA) from Merck (#810037); Human recombinant furin from OriGene Technologies Inc. (#TP304279M); Conventional heparin (13.6 kDa - purity ≥95%) from a commercial batch of unfractionated sodium heparin from Laboratori Derivati Organici S.p.A. (#9041-08-1).

Techniques: Tube Formation Assay

Journal: npj Viruses

Article Title: K5 polysaccharides inhibit SARS-CoV-2 infection by preventing spike-proteolytic priming

doi: 10.1038/s44298-025-00163-4

Figure Lengend Snippet: VeroE6 cells or A549 ACE2+ cells were treated with increasing concentrations of heparin, K5, K5OSH, and K5NOSH. a , b The cells remained viable in the presence of heparin and the K5 compounds as evaluated by measuring the ATP levels. VeroE6 or A549 ACE2+ cells were infected with the B.1 c , d or Omicron BA.1 e , f isolates in the presence or the absence of increasing concentrations of heparin, K5, K5OSH, and K5NOSH. Infection was reduced in a concentration-dependent manner as shown by the percentage of plaque reduction compared to SARS-CoV-2 alone. Data are presented as the mean value ± standard error of three independent replicates. * P < 0.05; ** P < 0.005.

Article Snippet: Reagents and materials were used as received, unless otherwise mentioned, and were purchased from the following: Human recombinant SARS-CoV-2 Wuhan-Hu-1 spike His-Tag protein and RBD from Sino Biological (#40592-V08B); ACE2 from Acrobiosystem (#AC2-H52H8); Bovine Serum Albumin (BSA) from Merck (#810037); Human recombinant furin from OriGene Technologies Inc. (#TP304279M); Conventional heparin (13.6 kDa - purity ≥95%) from a commercial batch of unfractionated sodium heparin from Laboratori Derivati Organici S.p.A. (#9041-08-1).

Techniques: Infection, Concentration Assay