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