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Addgene inc human ace2 gene
Human Ace2 Gene, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 173 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+ace2+hace2/us12569467-177-4-10?v=Addgene+inc
Average 96 stars, based on 173 article reviews

human ace2 gene - by Bioz Stars, 2026-06

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Schematic overview of Trp catabolism. (A) Diet with Trp is degraded into 2 major segments. Ninety percent is absorbed and converted to N-formylkynurenine in the liver via 2 important enzymes, IDO-1/IDO-2 and TDO2. The remaining 10% is converted into indole. N-Formylkynurenine is converted to Kyn via (aryl)formamidase. However, N-formylkynurenine is converted to KYNA via (KAT-I/II/III). Kyn is converted to 3-HK via KMO. 3-HK is further degraded to XA and to 3-HAA, the latter reaction being catalyzed by KYNU. (B) Schematic presentation of the experimental design. (i) Humanized transgenic ACE2-expressing mice were inoculated intranasally with SARS-COV-2, and at a specified time period, the mice were euthanized, and organs (kidney, lung, and liver) and blood samples were harvested. Samples were subjected to targeted Trp metabolites. (ii) Serum samples from patients with COVID-19 were examined for: (1) Trp metabolites by LC-MS–based metabolomics; and (2) pooled serum (5%) exposure of human ECs, followed by AHR and TF activity assays. Enzymatic reactions are shown in italics. 3-HAO, 3-hydroxyanthranilate 3,4-dioxygenase; ACMSD, aminocarboxymuconate semialdehyde decarboxylase; AMO, anthranilate 3-monooxygenase; i.n., intranasal; KAT, kynurenine aminotransferase; PFU, plaque-forming units; QPRT, quinolinate phosphoribosyltransferase.

Journal: Blood Vessels, Thrombosis & Hemostasis

Article Title: Tryptophan metabolism reprogramming potentially contributes to the prothrombotic milieu in mice and humans with SARS-CoV-2 ∗

doi: 10.1016/j.bvth.2025.100128

Figure Lengend Snippet: Schematic overview of Trp catabolism. (A) Diet with Trp is degraded into 2 major segments. Ninety percent is absorbed and converted to N-formylkynurenine in the liver via 2 important enzymes, IDO-1/IDO-2 and TDO2. The remaining 10% is converted into indole. N-Formylkynurenine is converted to Kyn via (aryl)formamidase. However, N-formylkynurenine is converted to KYNA via (KAT-I/II/III). Kyn is converted to 3-HK via KMO. 3-HK is further degraded to XA and to 3-HAA, the latter reaction being catalyzed by KYNU. (B) Schematic presentation of the experimental design. (i) Humanized transgenic ACE2-expressing mice were inoculated intranasally with SARS-COV-2, and at a specified time period, the mice were euthanized, and organs (kidney, lung, and liver) and blood samples were harvested. Samples were subjected to targeted Trp metabolites. (ii) Serum samples from patients with COVID-19 were examined for: (1) Trp metabolites by LC-MS–based metabolomics; and (2) pooled serum (5%) exposure of human ECs, followed by AHR and TF activity assays. Enzymatic reactions are shown in italics. 3-HAO, 3-hydroxyanthranilate 3,4-dioxygenase; ACMSD, aminocarboxymuconate semialdehyde decarboxylase; AMO, anthranilate 3-monooxygenase; i.n., intranasal; KAT, kynurenine aminotransferase; PFU, plaque-forming units; QPRT, quinolinate phosphoribosyltransferase.

Article Snippet: Heterozygous transgenic humanized ACE2 (K18-hACE2) mice (strain: 2B6.Cg-Tg(K18-ACE2)2Prlmn/J; mixed-sex) were obtained from The Jackson Laboratory (Bar Harbor, ME).

Techniques: Transgenic Assay, Expressing, Liquid Chromatography with Mass Spectroscopy, Activity Assay

TDO regulation in liver of humanized transgenic ACE2 mice infected with SARS-CoV-2. (A) Mice were infected with SARS-CoV-2, and paraffin-embedded liver sections were made at 2 dpi, 4 dpi, and 7 dpi and screened for TDO-1 immunoreactivity (n = 3). (i) Mice exposed to normal saline that serves as a mock control; (ii) 2 dpi; (iii) 4 dpi; and (iv) 7 dpi. Scale bar, 100 μm; the inserts depict the zoomed-in images. (B) Data were quantified and plotted as integrated density per micrometer. Data were analyzed using 1-way analysis of variance (ANOVA) followed by a multiple comparison test (∗ P < .05).

Journal: Blood Vessels, Thrombosis & Hemostasis

Article Title: Tryptophan metabolism reprogramming potentially contributes to the prothrombotic milieu in mice and humans with SARS-CoV-2 ∗

doi: 10.1016/j.bvth.2025.100128

Figure Lengend Snippet: TDO regulation in liver of humanized transgenic ACE2 mice infected with SARS-CoV-2. (A) Mice were infected with SARS-CoV-2, and paraffin-embedded liver sections were made at 2 dpi, 4 dpi, and 7 dpi and screened for TDO-1 immunoreactivity (n = 3). (i) Mice exposed to normal saline that serves as a mock control; (ii) 2 dpi; (iii) 4 dpi; and (iv) 7 dpi. Scale bar, 100 μm; the inserts depict the zoomed-in images. (B) Data were quantified and plotted as integrated density per micrometer. Data were analyzed using 1-way analysis of variance (ANOVA) followed by a multiple comparison test (∗ P < .05).

Article Snippet: Heterozygous transgenic humanized ACE2 (K18-hACE2) mice (strain: 2B6.Cg-Tg(K18-ACE2)2Prlmn/J; mixed-sex) were obtained from The Jackson Laboratory (Bar Harbor, ME).

Techniques: Transgenic Assay, Infection, Saline, Control, Comparison

IDO-1, KMO, and KYNU expression in renal tubules of humanized transgenic ACE2 mice infected with SARS-CoV-2. Mice were infected with SARS-CoV-2, and paraffin-embedded kidney sections were made at 2 dpi, 4 dpi, and 7 dpi and screened for IDO-1 (A), KMO (C), and KYNU immunoreactivity (n = 3) (E) in renal tubules. In panels A,C,E, mice were exposed to normal saline that serves as a mock control (i); the other images are from infected mice at 2 dpi (ii), 4 dpi (iii), and 7 dpi (iv). Scale bar, 100 μm; the inserts depict the zoomed-in images. (B,D,F) Data were quantified and plotted as integrated density per micrometer of respective markers shown in panels A,C,E. Data were analyzed using 1-way ANOVA followed by a multiple comparison test (∗∗ P < .01).

Journal: Blood Vessels, Thrombosis & Hemostasis

Article Title: Tryptophan metabolism reprogramming potentially contributes to the prothrombotic milieu in mice and humans with SARS-CoV-2 ∗

doi: 10.1016/j.bvth.2025.100128

Figure Lengend Snippet: IDO-1, KMO, and KYNU expression in renal tubules of humanized transgenic ACE2 mice infected with SARS-CoV-2. Mice were infected with SARS-CoV-2, and paraffin-embedded kidney sections were made at 2 dpi, 4 dpi, and 7 dpi and screened for IDO-1 (A), KMO (C), and KYNU immunoreactivity (n = 3) (E) in renal tubules. In panels A,C,E, mice were exposed to normal saline that serves as a mock control (i); the other images are from infected mice at 2 dpi (ii), 4 dpi (iii), and 7 dpi (iv). Scale bar, 100 μm; the inserts depict the zoomed-in images. (B,D,F) Data were quantified and plotted as integrated density per micrometer of respective markers shown in panels A,C,E. Data were analyzed using 1-way ANOVA followed by a multiple comparison test (∗∗ P < .01).

Article Snippet: Heterozygous transgenic humanized ACE2 (K18-hACE2) mice (strain: 2B6.Cg-Tg(K18-ACE2)2Prlmn/J; mixed-sex) were obtained from The Jackson Laboratory (Bar Harbor, ME).

Techniques: Expressing, Transgenic Assay, Infection, Saline, Control, Comparison

IDO-1, KMO, and KYNU expression in lungs of humanized transgenic ACE2 mice infected with SARS-CoV-2. Mice were infected with SARS-CoV-2, and paraffin-embedded lung sections were made at 2 dpi, 4 dpi, and 7 dpi and screened for IDO-1 (A), KMO (C), and KYNU immunoreactivity (n = 3) (E) in the lungs. In panels A,C,E, mice exposed to normal saline that serves as a mock control (i), 2 dpi (ii), 4 dpi (iii), and 7 dpi (iv). Scale bar, 100 μm; the inserts depict the zoomed-in images. (B,D,F) Data were quantified and plotted as integrated density per micrometer of respective markers shown in panels A,C,E. Data were analyzed using 1-way ANOVA followed by multiple comparison test (∗ P < .05; ∗∗∗∗ P < .0001).

Journal: Blood Vessels, Thrombosis & Hemostasis

Article Title: Tryptophan metabolism reprogramming potentially contributes to the prothrombotic milieu in mice and humans with SARS-CoV-2 ∗

doi: 10.1016/j.bvth.2025.100128

Figure Lengend Snippet: IDO-1, KMO, and KYNU expression in lungs of humanized transgenic ACE2 mice infected with SARS-CoV-2. Mice were infected with SARS-CoV-2, and paraffin-embedded lung sections were made at 2 dpi, 4 dpi, and 7 dpi and screened for IDO-1 (A), KMO (C), and KYNU immunoreactivity (n = 3) (E) in the lungs. In panels A,C,E, mice exposed to normal saline that serves as a mock control (i), 2 dpi (ii), 4 dpi (iii), and 7 dpi (iv). Scale bar, 100 μm; the inserts depict the zoomed-in images. (B,D,F) Data were quantified and plotted as integrated density per micrometer of respective markers shown in panels A,C,E. Data were analyzed using 1-way ANOVA followed by multiple comparison test (∗ P < .05; ∗∗∗∗ P < .0001).

Article Snippet: Heterozygous transgenic humanized ACE2 (K18-hACE2) mice (strain: 2B6.Cg-Tg(K18-ACE2)2Prlmn/J; mixed-sex) were obtained from The Jackson Laboratory (Bar Harbor, ME).

Techniques: Expressing, Transgenic Assay, Infection, Saline, Control, Comparison

LC-MS–based targeted metabolomics in sera, and AHR and TF expression in renal arteries and lung vasculature from K18-hACE2 mice (4 dpi). Serum samples from K18-hACE2 mice infected with SARS-CoV-2 were used for LC-MS–based metabolomic study. Separation was on Atlantis T3 C18 3 μm 50 × 2.1 mm column (Waters Corp, Milford, MA). A binary gradient consisting of solvent A (5 mmol/L ammonium acetate) and solvent B (methanol) and different gradients were applied (see “Methods” for specific details about each metabolite). MS of each metabolite was examined using an API 4000 triple quadrupole MS equipped with electrospray and an Agilent high-performance liquid chromatography (HPLC) G1312B pump, using plasma samples. (B-F) All 5 metabolites and their internal standards were infused into MS for scanning spectrum and determining fragmentation. All the metabolites were estimated from 4 dpi sera and compared with mock controls (n = 5 per group). Metabolites: Trp, Kyn, and kynurenic mouse sera; Student t test (∗ P < .05). K18-hACE2 mice were infected with SARS-CoV-2, and paraffin-embedded kidney sections (n = 3) were made at 4 dpi and stained for AHR in panel G and TF in panel I. CD31 was used in both the panels as an endothelial marker. Original magnifications, ×400 for panel G and ×200 for panel I. The yellow asterisk depicts the nuclear AHR expression. Data for AHR and TF in panels H,J were quantified and plotted as integrated density per micrometer. Data were analyzed using the Student t test (∗ P < .05). (K-N) Paraffin-embedded lung sections were prepared at 4 dpi and stained for AHR (K) and TF (M). CD31 was used in both the panels as an endothelial marker. Scale bar, 100 μm. The yellow asterisk depicts the nuclear AHR expression. Data in panels L,N were quantified and plotted as integrated density per micrometer. Data were analyzed using the Student t test (∗ P < .05; ∗∗ P < .01). DAPI, 4,ʹ6-diamidino-2-phenylindole; i.n., intranasal; L, lumen; ns, not significant; PFU, plaque-forming units.

Journal: Blood Vessels, Thrombosis & Hemostasis

Article Title: Tryptophan metabolism reprogramming potentially contributes to the prothrombotic milieu in mice and humans with SARS-CoV-2 ∗

doi: 10.1016/j.bvth.2025.100128

Figure Lengend Snippet: LC-MS–based targeted metabolomics in sera, and AHR and TF expression in renal arteries and lung vasculature from K18-hACE2 mice (4 dpi). Serum samples from K18-hACE2 mice infected with SARS-CoV-2 were used for LC-MS–based metabolomic study. Separation was on Atlantis T3 C18 3 μm 50 × 2.1 mm column (Waters Corp, Milford, MA). A binary gradient consisting of solvent A (5 mmol/L ammonium acetate) and solvent B (methanol) and different gradients were applied (see “Methods” for specific details about each metabolite). MS of each metabolite was examined using an API 4000 triple quadrupole MS equipped with electrospray and an Agilent high-performance liquid chromatography (HPLC) G1312B pump, using plasma samples. (B-F) All 5 metabolites and their internal standards were infused into MS for scanning spectrum and determining fragmentation. All the metabolites were estimated from 4 dpi sera and compared with mock controls (n = 5 per group). Metabolites: Trp, Kyn, and kynurenic mouse sera; Student t test (∗ P < .05). K18-hACE2 mice were infected with SARS-CoV-2, and paraffin-embedded kidney sections (n = 3) were made at 4 dpi and stained for AHR in panel G and TF in panel I. CD31 was used in both the panels as an endothelial marker. Original magnifications, ×400 for panel G and ×200 for panel I. The yellow asterisk depicts the nuclear AHR expression. Data for AHR and TF in panels H,J were quantified and plotted as integrated density per micrometer. Data were analyzed using the Student t test (∗ P < .05). (K-N) Paraffin-embedded lung sections were prepared at 4 dpi and stained for AHR (K) and TF (M). CD31 was used in both the panels as an endothelial marker. Scale bar, 100 μm. The yellow asterisk depicts the nuclear AHR expression. Data in panels L,N were quantified and plotted as integrated density per micrometer. Data were analyzed using the Student t test (∗ P < .05; ∗∗ P < .01). DAPI, 4,ʹ6-diamidino-2-phenylindole; i.n., intranasal; L, lumen; ns, not significant; PFU, plaque-forming units.

Article Snippet: Heterozygous transgenic humanized ACE2 (K18-hACE2) mice (strain: 2B6.Cg-Tg(K18-ACE2)2Prlmn/J; mixed-sex) were obtained from The Jackson Laboratory (Bar Harbor, ME).

Techniques: Liquid Chromatography with Mass Spectroscopy, Expressing, Infection, Metabolomic, Solvent, Targeted Proteomics, High Performance Liquid Chromatography, Clinical Proteomics, Staining, Marker

Evolution, prevalence, and replicative kinetics of XBB.1.9 subvariants. ( A ) Evolutionary origins of the XBB.1.9 sublineages, including XBB.1.9.1, EG.5.1, and HK.3. Synonymous mutations in nucleotides and amino acid mutations are shown in bold and bold-italic font, respectively. ( B ) Prevalence of XBB.1.9.1 (blue), EG.5.1 (green), and HK.3 (orange) in China (CHN), the United States (USA), Europe (EUP), and the Republic of Korea (ROK) for 14 months from January 2023 (2023.01) to March 2024 (2024.03). ( C and D ). Replicative kinetics of BA.2 (dark red), XBB.1.9.1 (blue), EG.5.1 (green), and HK.3 (orange) in terms of viral titers (upper panel) and viral loads (lower panel) in Vero E6, Vero E6 TMPRSS2+ , HeLa hACE2+ , Huh-7, and Caco2 cells. Cells were infected at an MOI of 0.01. The significance of the differences in replication between BA.2 and XBB.1.9.1, EG.5.1, or HK.3 is indicated above the lines by the asterisks in colors corresponding to the individual viruses. The significance of the differences in replication between HK.3 and XBB.1.9.1 or EG.5.1 is indicated by gray or black asterisks below the lines. A detection reference (from a weakly positive sample, CT = 27.0) is represented by dashed lines. ( E ) Viability of HK.3-infected cells. Significance of viability differences between Vero E6 and Vero E6 TMPRSS2+ cells is revealed. ( F ) Relative RNA expressions of TMPRSS2 (left) and ACE2 (right). Significance of the differences in TMPRSS2 expression between Vero E6 and other cells and in ACE2 expression between HeLa hACE2+ cells and others is indicated. ( G ) Ratio of viral titers (upper panel) and viral loads (lower panel) in Vero E6 cells with high versus low TMPRSS2 expression. ( H ) Replicative kinetics of two HK.3 isolates. Statistical analyses were conducted using Student’s t -test. Significances: P < 0.05 (*), P < 0.01 (**), or P < 0.001 (***). Viral titer reflects the number of infectious viral particles (TCID 50 /mL), whereas viral load represents RNA replication levels (copy number of genomic RNA).

Journal: Journal of Virology

Article Title: Increased pathogenicity and transmission of SARS-CoV-2 Omicron XBB.1.9 subvariants, including HK.3 and EG.5.1, relative to BA.2

doi: 10.1128/jvi.01342-25

Figure Lengend Snippet: Evolution, prevalence, and replicative kinetics of XBB.1.9 subvariants. ( A ) Evolutionary origins of the XBB.1.9 sublineages, including XBB.1.9.1, EG.5.1, and HK.3. Synonymous mutations in nucleotides and amino acid mutations are shown in bold and bold-italic font, respectively. ( B ) Prevalence of XBB.1.9.1 (blue), EG.5.1 (green), and HK.3 (orange) in China (CHN), the United States (USA), Europe (EUP), and the Republic of Korea (ROK) for 14 months from January 2023 (2023.01) to March 2024 (2024.03). ( C and D ). Replicative kinetics of BA.2 (dark red), XBB.1.9.1 (blue), EG.5.1 (green), and HK.3 (orange) in terms of viral titers (upper panel) and viral loads (lower panel) in Vero E6, Vero E6 TMPRSS2+ , HeLa hACE2+ , Huh-7, and Caco2 cells. Cells were infected at an MOI of 0.01. The significance of the differences in replication between BA.2 and XBB.1.9.1, EG.5.1, or HK.3 is indicated above the lines by the asterisks in colors corresponding to the individual viruses. The significance of the differences in replication between HK.3 and XBB.1.9.1 or EG.5.1 is indicated by gray or black asterisks below the lines. A detection reference (from a weakly positive sample, CT = 27.0) is represented by dashed lines. ( E ) Viability of HK.3-infected cells. Significance of viability differences between Vero E6 and Vero E6 TMPRSS2+ cells is revealed. ( F ) Relative RNA expressions of TMPRSS2 (left) and ACE2 (right). Significance of the differences in TMPRSS2 expression between Vero E6 and other cells and in ACE2 expression between HeLa hACE2+ cells and others is indicated. ( G ) Ratio of viral titers (upper panel) and viral loads (lower panel) in Vero E6 cells with high versus low TMPRSS2 expression. ( H ) Replicative kinetics of two HK.3 isolates. Statistical analyses were conducted using Student’s t -test. Significances: P < 0.05 (*), P < 0.01 (**), or P < 0.001 (***). Viral titer reflects the number of infectious viral particles (TCID 50 /mL), whereas viral load represents RNA replication levels (copy number of genomic RNA).

Article Snippet: The hACE2 protein with an Fc tag (Acro Biosystems) was immobilized onto the sample flow cell of the sensor chip.

Techniques: Infection, Expressing

Characteristics of the spikes of XBB.1.9 subvariants. ( A ) Spike-mediated infection determined by pseudovirus assays. XBB.1-S of the XBB.1 lineage and XBB.1-P of the XBB.1.9 lineage were included. The infection efficiency of BA.2 has been set to 1 to show relative infectivity. ( B ) Spike-mediated cell‒cell fusion based on luciferase activity. BA.2 (dark red), XBB.1-P (blue), EG.5.1 (green), and HK.3 (orange) are indicated by solid lines. D614G (pink), XBB.1-S (purple), and a negative control (N.C. in gray) are indicated by dotted lines. The significance of the differences between XBB.1 variants and BA.2 is indicated in colors corresponding to the individual XBB variants, which are placed within black rectangles by the asterisks, respectively. ( C ) Spike-mediated syncytia formation (scale bar: 400 µm). ( D ) The proteolytic processing of spike protein was analyzed in authentic SARS-CoV-2 virions propagated in Vero E6 TMPRSS2+ cells, including the ancestral strain (BJ05P14), Delta, and Omicron subvariants (BA.2, XBB.1.9.1, EG.5.1, and HK.3). Relative spike protein expression levels of BA.2, XBB.1.9.1, EG.5.1, and HK.3 virions were determined (a representative result) with an exposure time of 1 ms (left). The ratio of S2 subunit bands to full-length S protein (S2/S) was quantified (three biological replicates) using ImageJ/Fiji software (right). The ratio of BA.2 has been set to 1. ( E ) Purification of XBB.1-S, XBB.1-P, EG.5.1, and HK.3 spikes. ( F ) Comparison of the binding affinities of the XBB.1 spikes to hACE2. SPR characterization of the spike includes XBB.1-S, XBB.1-P, EG.5.1, and HK.3 interacting with hACE2. The dissociation constant is revealed above the lines. Sensorgrams depict different concentrations of ligands. Statistical analyses were conducted using Student’s t -test. Significances: P < 0.05 (*), P < 0.01 (**), or P < 0.001 (***).

Journal: Journal of Virology

Article Title: Increased pathogenicity and transmission of SARS-CoV-2 Omicron XBB.1.9 subvariants, including HK.3 and EG.5.1, relative to BA.2

doi: 10.1128/jvi.01342-25

Figure Lengend Snippet: Characteristics of the spikes of XBB.1.9 subvariants. ( A ) Spike-mediated infection determined by pseudovirus assays. XBB.1-S of the XBB.1 lineage and XBB.1-P of the XBB.1.9 lineage were included. The infection efficiency of BA.2 has been set to 1 to show relative infectivity. ( B ) Spike-mediated cell‒cell fusion based on luciferase activity. BA.2 (dark red), XBB.1-P (blue), EG.5.1 (green), and HK.3 (orange) are indicated by solid lines. D614G (pink), XBB.1-S (purple), and a negative control (N.C. in gray) are indicated by dotted lines. The significance of the differences between XBB.1 variants and BA.2 is indicated in colors corresponding to the individual XBB variants, which are placed within black rectangles by the asterisks, respectively. ( C ) Spike-mediated syncytia formation (scale bar: 400 µm). ( D ) The proteolytic processing of spike protein was analyzed in authentic SARS-CoV-2 virions propagated in Vero E6 TMPRSS2+ cells, including the ancestral strain (BJ05P14), Delta, and Omicron subvariants (BA.2, XBB.1.9.1, EG.5.1, and HK.3). Relative spike protein expression levels of BA.2, XBB.1.9.1, EG.5.1, and HK.3 virions were determined (a representative result) with an exposure time of 1 ms (left). The ratio of S2 subunit bands to full-length S protein (S2/S) was quantified (three biological replicates) using ImageJ/Fiji software (right). The ratio of BA.2 has been set to 1. ( E ) Purification of XBB.1-S, XBB.1-P, EG.5.1, and HK.3 spikes. ( F ) Comparison of the binding affinities of the XBB.1 spikes to hACE2. SPR characterization of the spike includes XBB.1-S, XBB.1-P, EG.5.1, and HK.3 interacting with hACE2. The dissociation constant is revealed above the lines. Sensorgrams depict different concentrations of ligands. Statistical analyses were conducted using Student’s t -test. Significances: P < 0.05 (*), P < 0.01 (**), or P < 0.001 (***).

Article Snippet: The hACE2 protein with an Fc tag (Acro Biosystems) was immobilized onto the sample flow cell of the sensor chip.

Techniques: Infection, Luciferase, Activity Assay, Negative Control, Expressing, Software, Purification, Comparison, Binding Assay

Competitive fitness of XBB.1.9.1 and EG.5.1/HK.3 in wild-type hamsters. ( A ) Relative infection tropism of spikes. The infectivity ratio of ghACE2 to hACE2 is determined as tropism. ( B ) Flow chart of competitive fitness. ( C and D ) A mixture of XBB.1.9.1 and EG.5.1 ( C ) or HK.3 ( D ) at viral titer ratios of 1:1 (upper panel) or 1:3 (lower panel) was inoculated into hamsters. The RNA proportion of XBB.1.9.1 in the mixture was shown by numbers in the bars. Firstly, the RNA proportion of XBB.1.9.1 in initial inoculum was 80.8% or 54.5% ( C ) and 94.9% or 90.1% ( D ) which was displayed on the right of the initial proportion (yellow number). Secondly, the RNA proportion of XBB.1.9.1 in tissue samples (3 DPI) was shown in the bars (white number) below the horizontal of each figure grouping. The area in the bar means the RNA proportions of XBB.1.9.1 (blue) and EG.5.1 (green) or HK.3 (orange). Tissue samples are the lung and turbinate: lung (left) and turbinate (right).

Journal: Journal of Virology

Article Title: Increased pathogenicity and transmission of SARS-CoV-2 Omicron XBB.1.9 subvariants, including HK.3 and EG.5.1, relative to BA.2

doi: 10.1128/jvi.01342-25

Figure Lengend Snippet: Competitive fitness of XBB.1.9.1 and EG.5.1/HK.3 in wild-type hamsters. ( A ) Relative infection tropism of spikes. The infectivity ratio of ghACE2 to hACE2 is determined as tropism. ( B ) Flow chart of competitive fitness. ( C and D ) A mixture of XBB.1.9.1 and EG.5.1 ( C ) or HK.3 ( D ) at viral titer ratios of 1:1 (upper panel) or 1:3 (lower panel) was inoculated into hamsters. The RNA proportion of XBB.1.9.1 in the mixture was shown by numbers in the bars. Firstly, the RNA proportion of XBB.1.9.1 in initial inoculum was 80.8% or 54.5% ( C ) and 94.9% or 90.1% ( D ) which was displayed on the right of the initial proportion (yellow number). Secondly, the RNA proportion of XBB.1.9.1 in tissue samples (3 DPI) was shown in the bars (white number) below the horizontal of each figure grouping. The area in the bar means the RNA proportions of XBB.1.9.1 (blue) and EG.5.1 (green) or HK.3 (orange). Tissue samples are the lung and turbinate: lung (left) and turbinate (right).

Article Snippet: The hACE2 protein with an Fc tag (Acro Biosystems) was immobilized onto the sample flow cell of the sensor chip.

Techniques: Infection

In vivo virological characteristics of XBB.1.9 subvariants in K18-hACE2 hamsters. K18-hACE2 hamsters were intranasally inoculated with BA.2, XBB.1.9.1, EG.5.1, or HK.3. Four hamsters per group were used to measure the various parameters ( A, B, and C ). Four hamsters per group were euthanized at 3 DPI and used for data collection ( D–H ). The data (in A to E) of the mock, BA.2, XBB.1.9.1, EG.5.1, and HK.3 groups are shown in gray, red, blue, green, and orange, respectively (as shown in panel A ). ( A ) Body weights of the infected hamsters. Significant differences between the mock group and each infected group are revealed above the lines using asterisks in the colors corresponding to the respective infected group. ( B ) Percentage survival of the infected hamsters. Survival differences between multiple XBB.1.9 variants and BA.2 were analyzed using a Log-rank (Mantel-Cox) test with significance displayed in colors corresponding to the individual XBB.1.9 variant. ( C ) Viral loads in the nasal lavages of hamsters. The viral load baseline is indicated by dotted gray lines. ( D and E ) Radar chart of pathology ( D ) and pathology scores ( E ) of the infected lungs of hACE2 hamsters. The average of BA.2 (dark red), XBB.1.9.1 (blue), EG.5.1 (green), and HK.3 (orange) infected hamster (of 3–4 individuals) was indicated. ( F ) H&E staining and IHC images of the lungs of the infected hamsters. The lungs of two infected individuals in each group, namely, repetition 1 (REP1) and repetition 2 (REP2), are shown. The time point of tissue samples corresponds to 3 DPI. The scale bar represents 100 µm. ( G and H ) Viral titers ( G ) and viral loads ( H ) in the lungs (dark red) or turbinates (gray) of the infected hamsters. Statistical analyses were conducted using Student’s t -test. Significances: P < 0.05 (*), P < 0.01 (**), or P < 0.001 (***).

Journal: Journal of Virology

Article Title: Increased pathogenicity and transmission of SARS-CoV-2 Omicron XBB.1.9 subvariants, including HK.3 and EG.5.1, relative to BA.2

doi: 10.1128/jvi.01342-25

Figure Lengend Snippet: In vivo virological characteristics of XBB.1.9 subvariants in K18-hACE2 hamsters. K18-hACE2 hamsters were intranasally inoculated with BA.2, XBB.1.9.1, EG.5.1, or HK.3. Four hamsters per group were used to measure the various parameters ( A, B, and C ). Four hamsters per group were euthanized at 3 DPI and used for data collection ( D–H ). The data (in A to E) of the mock, BA.2, XBB.1.9.1, EG.5.1, and HK.3 groups are shown in gray, red, blue, green, and orange, respectively (as shown in panel A ). ( A ) Body weights of the infected hamsters. Significant differences between the mock group and each infected group are revealed above the lines using asterisks in the colors corresponding to the respective infected group. ( B ) Percentage survival of the infected hamsters. Survival differences between multiple XBB.1.9 variants and BA.2 were analyzed using a Log-rank (Mantel-Cox) test with significance displayed in colors corresponding to the individual XBB.1.9 variant. ( C ) Viral loads in the nasal lavages of hamsters. The viral load baseline is indicated by dotted gray lines. ( D and E ) Radar chart of pathology ( D ) and pathology scores ( E ) of the infected lungs of hACE2 hamsters. The average of BA.2 (dark red), XBB.1.9.1 (blue), EG.5.1 (green), and HK.3 (orange) infected hamster (of 3–4 individuals) was indicated. ( F ) H&E staining and IHC images of the lungs of the infected hamsters. The lungs of two infected individuals in each group, namely, repetition 1 (REP1) and repetition 2 (REP2), are shown. The time point of tissue samples corresponds to 3 DPI. The scale bar represents 100 µm. ( G and H ) Viral titers ( G ) and viral loads ( H ) in the lungs (dark red) or turbinates (gray) of the infected hamsters. Statistical analyses were conducted using Student’s t -test. Significances: P < 0.05 (*), P < 0.01 (**), or P < 0.001 (***).

Article Snippet: The hACE2 protein with an Fc tag (Acro Biosystems) was immobilized onto the sample flow cell of the sensor chip.

Techniques: In Vivo, Infection, Variant Assay, Staining

a Kinetic analysis of B07, B09, and B10 VHHs on hACE2-Fc by BioLayer Interferometry (BLI) using various concentrations of VHHs (as indicated on the figure). AHC biosensors were used to immobilize hACE2-Fc. K D of each VHHs is indicated. One representative experiment out of two. b Epitope mapping. sACE2-Fc (5 µg/mL) was immobilized onto the AHC biosensors. After a baseline step, a first VHH was applied (VHH1; 5 µg/mL). The sensor was dipped in a mixture of VHH1 and the competitor VHH2 at the same concentration (5 µg/mL). An anti-IgM specific VHH was used as control (ctl). (Top) Immobilized sACE2 pre-bound to B07 (VHH1) and VHH2 being B09, B10, or IgM (ctl). (Middle) Immobilized sACE2 pre-bound to B09 (VHH1) and VHH2 being B07, B10, or IgM (ctl). (Bottom) Immobilized sACE2 pre-bound to B10 (VHH1) and VHH2 being B07, B09, or IgM (ctl). Source Data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Targeting ACE2 with a camelid antibody inhibits SARS-CoV-2 binding and has protective effects in vivo

doi: 10.1038/s41467-025-65144-w

Figure Lengend Snippet: a Kinetic analysis of B07, B09, and B10 VHHs on hACE2-Fc by BioLayer Interferometry (BLI) using various concentrations of VHHs (as indicated on the figure). AHC biosensors were used to immobilize hACE2-Fc. K D of each VHHs is indicated. One representative experiment out of two. b Epitope mapping. sACE2-Fc (5 µg/mL) was immobilized onto the AHC biosensors. After a baseline step, a first VHH was applied (VHH1; 5 µg/mL). The sensor was dipped in a mixture of VHH1 and the competitor VHH2 at the same concentration (5 µg/mL). An anti-IgM specific VHH was used as control (ctl). (Top) Immobilized sACE2 pre-bound to B07 (VHH1) and VHH2 being B09, B10, or IgM (ctl). (Middle) Immobilized sACE2 pre-bound to B09 (VHH1) and VHH2 being B07, B10, or IgM (ctl). (Bottom) Immobilized sACE2 pre-bound to B10 (VHH1) and VHH2 being B07, B09, or IgM (ctl). Source Data are provided as a Source Data file.

Article Snippet: The construct human ACE2 (hACE2) was obtained by substitution of the sequence encoding mCardinal from mCardinal-C1 plasmid (addgene, #54799) with hACE2 from the pLenti6-attB-hACE2-BSD .

Techniques: Concentration Assay, Control

Cells were incubated with the different VHHs (10 µg/mL), stained with an anti-myc antibody and a AF488-conjugated anti-mouse antibody, before being analyzed by flow cytometry. a B07, B09, B10 binding efficacy on HEK293-hACE2-expressing cells. Data are means ± SD of three independent experiments. Parental HEK293 cells were used as control. b B07, B09, B10 binding efficacy on cells expressing endogenous hACE2 (IGROV-1). Anti-IgE VHH was used as a control (ctl). Data are means ± SD, n = 6 (ctl, B07) or 3 (B09, B10) independent experiments. c Fluorescence diagram overlays: B07, B09, B10 efficacy on cells expressing exogenous (HEK293-hACE2) or endogenous hACE2 (IGROV-1). Background (blue) corresponds to the fluorescence intensity obtained on parental cells (HEK293) or using a VHH control (anti-IgE VHH) (IGROV-1). Source Data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Targeting ACE2 with a camelid antibody inhibits SARS-CoV-2 binding and has protective effects in vivo

doi: 10.1038/s41467-025-65144-w

Figure Lengend Snippet: Cells were incubated with the different VHHs (10 µg/mL), stained with an anti-myc antibody and a AF488-conjugated anti-mouse antibody, before being analyzed by flow cytometry. a B07, B09, B10 binding efficacy on HEK293-hACE2-expressing cells. Data are means ± SD of three independent experiments. Parental HEK293 cells were used as control. b B07, B09, B10 binding efficacy on cells expressing endogenous hACE2 (IGROV-1). Anti-IgE VHH was used as a control (ctl). Data are means ± SD, n = 6 (ctl, B07) or 3 (B09, B10) independent experiments. c Fluorescence diagram overlays: B07, B09, B10 efficacy on cells expressing exogenous (HEK293-hACE2) or endogenous hACE2 (IGROV-1). Background (blue) corresponds to the fluorescence intensity obtained on parental cells (HEK293) or using a VHH control (anti-IgE VHH) (IGROV-1). Source Data are provided as a Source Data file.

Techniques: Incubation, Staining, Flow Cytometry, Binding Assay, Expressing, Control, Fluorescence

hACE2 activity in the presence of VHHs B07, B09, B10 or absence (-) was assayed using the SensoLyte 390 ACE2 Activity Assay Kit, which measures fluorogenic peptide cleavage. a Soluble hACE2 (sACE2) activity in the presence of 10 µM VHHs. An ACE2 inhibitor, provided with the kit, was used as control (DX600, 1 µM). Data are means ± SD of three independent experiments. Paired T test two-sided: ns, nonsignificant; ** p = 0.0053. b VHHs B07, B09, B10 were used at different doses (one representative experiment out of three performed in duplicates. Mean values of the duplicate). The mock control (buffer) was arbitrarily positioned at 10 -14 . c The ACE2 inhibitor DX600 was used at different doses. Data are means ± SD of seven independent experiments. The mock control (buffer) was arbitrarily positioned at 10 -14 . d Cell surface hACE2 activity in the presence of saturating concentration of VHHs ( > 20 µM) or 1 µM DX600. Data are means ± SD of four independent experiments. Paired T test two-sided: ns, nonsignificant; ** p = 0.0070. Source Data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Targeting ACE2 with a camelid antibody inhibits SARS-CoV-2 binding and has protective effects in vivo

doi: 10.1038/s41467-025-65144-w

Figure Lengend Snippet: hACE2 activity in the presence of VHHs B07, B09, B10 or absence (-) was assayed using the SensoLyte 390 ACE2 Activity Assay Kit, which measures fluorogenic peptide cleavage. a Soluble hACE2 (sACE2) activity in the presence of 10 µM VHHs. An ACE2 inhibitor, provided with the kit, was used as control (DX600, 1 µM). Data are means ± SD of three independent experiments. Paired T test two-sided: ns, nonsignificant; ** p = 0.0053. b VHHs B07, B09, B10 were used at different doses (one representative experiment out of three performed in duplicates. Mean values of the duplicate). The mock control (buffer) was arbitrarily positioned at 10 -14 . c The ACE2 inhibitor DX600 was used at different doses. Data are means ± SD of seven independent experiments. The mock control (buffer) was arbitrarily positioned at 10 -14 . d Cell surface hACE2 activity in the presence of saturating concentration of VHHs ( > 20 µM) or 1 µM DX600. Data are means ± SD of four independent experiments. Paired T test two-sided: ns, nonsignificant; ** p = 0.0070. Source Data are provided as a Source Data file.

Techniques: Activity Assay, Control, Concentration Assay

a S-fuse assay. Inhibition by B07, B09, B10 VHHs after infection of S-Fuse cells (U2OS-hACE2 GFP1-10 and GFP11) with different SARS-CoV-2 variants (Delta B.1.617.2, BA.1, BQ.1.1, XBB.1.5, XBB1.16.1, EG.5.1.3, BA.2.86.1). Data are mean of two independent experiments. The dashed line indicates the limit of detection. An anti-IgM (or IgE) specific VHH was used as a control (ctl). IC50 values are indicated in the table. b Inhibition of 5 µg/mL spike binding to hACE2-expressing HEK293 cells by increasing concentrations of VHHs B07, B09 or B10. HEK293-hACE2 cells pretreated or not with the VHHs and incubated with soluble spike (S) protein (ancestral spike) were stained with an anti-S antibody. Results were normalized for nonspecific (0%) and specific binding in the absence of inhibitor (100%). Experiments were fitted to a one-site competitive binding model. Data are mean ± SD of three (B07, B09) independent experiments or mean of two independent experiments (B10). c Displacement of S (ancestral spike) binding by B07 (left) or B09 (right) for various concentrations of S (5 µg/mL, 8 µg/mL, 10 µg/mL). Experiments were carried out as in ( b ) (one representative experiment out of two - 10 µg/mL, or three - 5 µg/mL, 8 µg/mL). d Relationship between the observed IC50 values for B07 or B09 displacement of S binding and initial concentration of S (ancestral spike). Results represent means ± SD of three independent experiments (5 µg/mL, 8 µg/mL) or means of two independent experiments (10 µg/mL). Linear regression analysis: R B07 = 0.9005; R B09 = 0.7896. Source Data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Targeting ACE2 with a camelid antibody inhibits SARS-CoV-2 binding and has protective effects in vivo

doi: 10.1038/s41467-025-65144-w

Figure Lengend Snippet: a S-fuse assay. Inhibition by B07, B09, B10 VHHs after infection of S-Fuse cells (U2OS-hACE2 GFP1-10 and GFP11) with different SARS-CoV-2 variants (Delta B.1.617.2, BA.1, BQ.1.1, XBB.1.5, XBB1.16.1, EG.5.1.3, BA.2.86.1). Data are mean of two independent experiments. The dashed line indicates the limit of detection. An anti-IgM (or IgE) specific VHH was used as a control (ctl). IC50 values are indicated in the table. b Inhibition of 5 µg/mL spike binding to hACE2-expressing HEK293 cells by increasing concentrations of VHHs B07, B09 or B10. HEK293-hACE2 cells pretreated or not with the VHHs and incubated with soluble spike (S) protein (ancestral spike) were stained with an anti-S antibody. Results were normalized for nonspecific (0%) and specific binding in the absence of inhibitor (100%). Experiments were fitted to a one-site competitive binding model. Data are mean ± SD of three (B07, B09) independent experiments or mean of two independent experiments (B10). c Displacement of S (ancestral spike) binding by B07 (left) or B09 (right) for various concentrations of S (5 µg/mL, 8 µg/mL, 10 µg/mL). Experiments were carried out as in ( b ) (one representative experiment out of two - 10 µg/mL, or three - 5 µg/mL, 8 µg/mL). d Relationship between the observed IC50 values for B07 or B09 displacement of S binding and initial concentration of S (ancestral spike). Results represent means ± SD of three independent experiments (5 µg/mL, 8 µg/mL) or means of two independent experiments (10 µg/mL). Linear regression analysis: R B07 = 0.9005; R B09 = 0.7896. Source Data are provided as a Source Data file.

Techniques: Inhibition, Infection, Control, Binding Assay, Expressing, Incubation, Staining, Concentration Assay

a Crystal structure of hACE2 (light blue) in complex with B07 (purple) and B10 (yellow) (PDB: 9R19). b Surface representation of the complex between hACE2 (light blue) and B07 (purple) rotated to 90 ° C from a . For clarity B10 was not displayed. c Open-book representation and footprints (in white) of B07 on hACE2 (left) and hACE2 on B07 (right). The buried surfaces (BSA) are indicated under each surface. Residues of hACE2 in red correspond to those for which alanine substitutions compromise B07 VHH binding to hACE2 mutants ( f ). d B07 complementarity determining regions (CDRs) colored in green (CDR1), blue (CDR2), and red (CDR3). hACE2 footprint on B07 is represented. e Hydrogen bonds and salt bridges (yellow dashed lines) between hACE2 and B07. f Impact of hACE2 alanine substitution on B07 binding. Cells were transfected with myc tagged or untagged hACE2 alanine mutants, incubated with an anti-myc antibody or B07 (10 µg/mL), respectively and stained with a mouse anti-myc antibody and a AF647-conjugated anti-mouse antibody before being analyzed by flow cytometry. Data are mean ± SD of four (anti-myc) or three (B07) independent experiments. Source Data are provided as a Source Data file. g Electrostatic potential mapped on the surface of the structure of hACE2 and B07. h Superimposition of the binding area of B07 (purple) and SARS-CoV-2 RBD (PDB: 6M0J, green) on hACE2 (light blue). i Footprints of the site of interaction of B07 and RBD on hACE2. The contours are colored in purple (B07) or in green (RBD). The common area is striped.

Journal: Nature Communications

Article Title: Targeting ACE2 with a camelid antibody inhibits SARS-CoV-2 binding and has protective effects in vivo

doi: 10.1038/s41467-025-65144-w

Figure Lengend Snippet: a Crystal structure of hACE2 (light blue) in complex with B07 (purple) and B10 (yellow) (PDB: 9R19). b Surface representation of the complex between hACE2 (light blue) and B07 (purple) rotated to 90 ° C from a . For clarity B10 was not displayed. c Open-book representation and footprints (in white) of B07 on hACE2 (left) and hACE2 on B07 (right). The buried surfaces (BSA) are indicated under each surface. Residues of hACE2 in red correspond to those for which alanine substitutions compromise B07 VHH binding to hACE2 mutants ( f ). d B07 complementarity determining regions (CDRs) colored in green (CDR1), blue (CDR2), and red (CDR3). hACE2 footprint on B07 is represented. e Hydrogen bonds and salt bridges (yellow dashed lines) between hACE2 and B07. f Impact of hACE2 alanine substitution on B07 binding. Cells were transfected with myc tagged or untagged hACE2 alanine mutants, incubated with an anti-myc antibody or B07 (10 µg/mL), respectively and stained with a mouse anti-myc antibody and a AF647-conjugated anti-mouse antibody before being analyzed by flow cytometry. Data are mean ± SD of four (anti-myc) or three (B07) independent experiments. Source Data are provided as a Source Data file. g Electrostatic potential mapped on the surface of the structure of hACE2 and B07. h Superimposition of the binding area of B07 (purple) and SARS-CoV-2 RBD (PDB: 6M0J, green) on hACE2 (light blue). i Footprints of the site of interaction of B07 and RBD on hACE2. The contours are colored in purple (B07) or in green (RBD). The common area is striped.

Techniques: Binding Assay, Transfection, Incubation, Staining, Flow Cytometry

a Kinetic analysis of VHH B07-Fc on hACE2-His by BioLayer Interferometry (BLI) using various concentrations of VHH B07-Fc (indicated on the figure). HIS2 biosensors were used to immobilize 1 µg/mL of hACE2-His. b VHH B07-Fc binding on cells expressing exogenous (HEK293-hACE2) or endogenous hACE2 (IGROV-1). Experiments were performed as in Fig. but using 0.1 µg/mL of VHH B07-Fc. Data are mean ± SD of six (HEK293) or four (IGROV-1) independent experiments. c Fluorescence diagram overlays as performed in Fig. . d hACE2, phalloidin (F-actin) and DAPI staining on primary human nasal epithelial cells (hNECs). Representative immunofluorescence staining (out of 4 independent experiments) of hACE2 (Red: B07-Fc staining) in combination with phalloidin (Cyan) and DAPI (Blue). Scale bars: ctl-Fc, 10 µm; B07-Fc, 5 µm. e Enzymatic activity of soluble hACE2 (sACE2) in the presence of VHH B07-Fc or absence using the SensoLyte 390 ACE2 Activity Assay Kit, as performed in Fig. . DX600 was an inhibitor control of the kit (one representative experiment out of two; mean values of duplicates). f Cell surface hACE2 activity in the presence of saturating concentration of VHHs (10 µM) or 1 µM DX600. Data are means ± SD of three independent experiments. Paired T test two-sided: ns, nonsignificant; ** p = 0.0070. g Inhibition of fusion (S-Fuse assay) by B07-Fc after infection of S-Fuse cells with different SARS-CoV-2 variants: Delta B.1.617.2, BA.1, BQ.1.1, XBB.1.5, XBB.1.16.1, EG.5.1.3, BA.2.86.1, JN.1.1, KP.3.3. The dashed line indicates the limit of detection. Data are mean of two independent experiments, except for BQ1.1 (three experiments). h Inhibition of infection quantified by counting SARS-CoV-2 nucleoprotein (N) positive cells. IGROV-1 cells were pre-incubated 1 h with serial dilutions of B07-Fc and infected with the indicated variants at 3 × 10 -2 infectious units per cell. Cells were stained with a pan-coronavirus anti-N antibody at day 1 pi. The percentage of inhibition is represented. One representative experiment performed in duplicates. i − k Impact of ACE2 genetic variation on VHH B07-Fc binding. i , j Cells were transfected with different ACE2 constructs ( i ) hACE2 polymorphism; ( j ) human, hamster, or mouse ACE2, incubated with B07-Fc (0.1 µg/mL or different doses) and stained with an AF647-conjugated anti-human antibody before being analyzed by flow cytometry. i Data are mean ± SD of independent experiments: n = 5 (S19P; I21T; K26R; T27A; E35K) or n = 4 (E37K; K68E; M82I, P84T) or n = 3 (H34N). j Data are mean ± SD of 5 (human ACE2) or 3 (hamster and mouse ACE2) independent experiments. k Kinetic analysis of B07-Fc on hamster ACE2-His by BioLayer Interferometry (BLI) using various concentrations of VHH B07-Fc (indicated on the figure). HIS2 biosensors were used to immobilize 2 µg/mL of His-ACE2. K D App is indicated. Source Data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Targeting ACE2 with a camelid antibody inhibits SARS-CoV-2 binding and has protective effects in vivo

doi: 10.1038/s41467-025-65144-w

Figure Lengend Snippet: a Kinetic analysis of VHH B07-Fc on hACE2-His by BioLayer Interferometry (BLI) using various concentrations of VHH B07-Fc (indicated on the figure). HIS2 biosensors were used to immobilize 1 µg/mL of hACE2-His. b VHH B07-Fc binding on cells expressing exogenous (HEK293-hACE2) or endogenous hACE2 (IGROV-1). Experiments were performed as in Fig. but using 0.1 µg/mL of VHH B07-Fc. Data are mean ± SD of six (HEK293) or four (IGROV-1) independent experiments. c Fluorescence diagram overlays as performed in Fig. . d hACE2, phalloidin (F-actin) and DAPI staining on primary human nasal epithelial cells (hNECs). Representative immunofluorescence staining (out of 4 independent experiments) of hACE2 (Red: B07-Fc staining) in combination with phalloidin (Cyan) and DAPI (Blue). Scale bars: ctl-Fc, 10 µm; B07-Fc, 5 µm. e Enzymatic activity of soluble hACE2 (sACE2) in the presence of VHH B07-Fc or absence using the SensoLyte 390 ACE2 Activity Assay Kit, as performed in Fig. . DX600 was an inhibitor control of the kit (one representative experiment out of two; mean values of duplicates). f Cell surface hACE2 activity in the presence of saturating concentration of VHHs (10 µM) or 1 µM DX600. Data are means ± SD of three independent experiments. Paired T test two-sided: ns, nonsignificant; ** p = 0.0070. g Inhibition of fusion (S-Fuse assay) by B07-Fc after infection of S-Fuse cells with different SARS-CoV-2 variants: Delta B.1.617.2, BA.1, BQ.1.1, XBB.1.5, XBB.1.16.1, EG.5.1.3, BA.2.86.1, JN.1.1, KP.3.3. The dashed line indicates the limit of detection. Data are mean of two independent experiments, except for BQ1.1 (three experiments). h Inhibition of infection quantified by counting SARS-CoV-2 nucleoprotein (N) positive cells. IGROV-1 cells were pre-incubated 1 h with serial dilutions of B07-Fc and infected with the indicated variants at 3 × 10 -2 infectious units per cell. Cells were stained with a pan-coronavirus anti-N antibody at day 1 pi. The percentage of inhibition is represented. One representative experiment performed in duplicates. i − k Impact of ACE2 genetic variation on VHH B07-Fc binding. i , j Cells were transfected with different ACE2 constructs ( i ) hACE2 polymorphism; ( j ) human, hamster, or mouse ACE2, incubated with B07-Fc (0.1 µg/mL or different doses) and stained with an AF647-conjugated anti-human antibody before being analyzed by flow cytometry. i Data are mean ± SD of independent experiments: n = 5 (S19P; I21T; K26R; T27A; E35K) or n = 4 (E37K; K68E; M82I, P84T) or n = 3 (H34N). j Data are mean ± SD of 5 (human ACE2) or 3 (hamster and mouse ACE2) independent experiments. k Kinetic analysis of B07-Fc on hamster ACE2-His by BioLayer Interferometry (BLI) using various concentrations of VHH B07-Fc (indicated on the figure). HIS2 biosensors were used to immobilize 2 µg/mL of His-ACE2. K D App is indicated. Source Data are provided as a Source Data file.

Techniques: Binding Assay, Expressing, Fluorescence, Staining, Immunofluorescence, Activity Assay, Control, Concentration Assay, Inhibition, Infection, Incubation, Transfection, Construct, Flow Cytometry

a Schematic diagram showing the experimental design of B07-Fc prophylaxis in XBB.1.5 infected K18-hACE2 mice. Animals received intranasally (i.n.) 7 mg/kg VHH-B07-Fc (B07-Fc) or 7 mg/kg VHH-Fc ctl (ctl-Fc). Twenty-four hours later, they were infected with 10 5 PFU XBB.1.5. SARS-CoV-2 intranasally (i.n.). Three days post-infection, different tissues were collected for analysis. b Genomic RNA load measured by RT-qPCR of SARS-CoV-2 in lung. c SARS-CoV-2 infectious particles measured by conventional plaque assays. d Tubulin, phalloidin (F-actin), SARS-CoV-2 nucleocapsid (N) (viral replication) and DAPI staining on nasal respiratory epithelium, olfactory epithelium, and lung bronchioli extracted from K18-hACE2 uninfected mice, and animals that received VHH-Fc (Ctl-Fc or B07-Fc). Representative immunofluorescence staining of Tubulin (Green) in combination with phalloidin (Cyan), SARS-CoV-2 N protein (Red), and DAPI (Blue). This was performed on nasal, olfactory, and pulmonary epithelium of two VHH ctl-treated mice and four B07-Fc treated mice. Scale bars: 10 µm. e Relationship between viral genomic RNA load measured in the lung by RT-qPCR and B07-Fc concentration quantified in the lung by ELISA. f Schematic diagram showing the experimental design of B07-Fc prophylaxis in D614G infected K18-hACE2 mice. Animals received intranasally (i.n.) 5 mg/kg VHH-B07-Fc (B07-Fc) or 5 mg/kg VHH-Fc ctl (ctl-Fc). Twenty-four hours later, they were infected with 10 4 PFU D614G SARS-CoV-2 i.n. Mice were clinically monitored between days 3 to 6. Mice were euthanized on day 6 and lungs were collected for analysis. g Body weight measured over time post-infection in ctl-Fc ( n = 5) and B07-Fc-treated groups. Mice of the B07-Fc group were split into VHH_high ( n = 4) and VHH_low ( n = 6) according to the B07-Fc concentration measured in the lung on day 6 p.i. (higher or lower than 200 ng/g of lung, respectively). Data are presented as mean values +/- SD. Linear mixed model two sided (no adjustment for multiple comparisons): ns, nonsignificant (p = 0.26); ** p = 0.00378; *** p = 0.000396. h Relationship between body weight loss and B07-Fc concentration quantified in the lung by ELISA on day 6 p.i. Source Data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Targeting ACE2 with a camelid antibody inhibits SARS-CoV-2 binding and has protective effects in vivo

doi: 10.1038/s41467-025-65144-w

Figure Lengend Snippet: a Schematic diagram showing the experimental design of B07-Fc prophylaxis in XBB.1.5 infected K18-hACE2 mice. Animals received intranasally (i.n.) 7 mg/kg VHH-B07-Fc (B07-Fc) or 7 mg/kg VHH-Fc ctl (ctl-Fc). Twenty-four hours later, they were infected with 10 5 PFU XBB.1.5. SARS-CoV-2 intranasally (i.n.). Three days post-infection, different tissues were collected for analysis. b Genomic RNA load measured by RT-qPCR of SARS-CoV-2 in lung. c SARS-CoV-2 infectious particles measured by conventional plaque assays. d Tubulin, phalloidin (F-actin), SARS-CoV-2 nucleocapsid (N) (viral replication) and DAPI staining on nasal respiratory epithelium, olfactory epithelium, and lung bronchioli extracted from K18-hACE2 uninfected mice, and animals that received VHH-Fc (Ctl-Fc or B07-Fc). Representative immunofluorescence staining of Tubulin (Green) in combination with phalloidin (Cyan), SARS-CoV-2 N protein (Red), and DAPI (Blue). This was performed on nasal, olfactory, and pulmonary epithelium of two VHH ctl-treated mice and four B07-Fc treated mice. Scale bars: 10 µm. e Relationship between viral genomic RNA load measured in the lung by RT-qPCR and B07-Fc concentration quantified in the lung by ELISA. f Schematic diagram showing the experimental design of B07-Fc prophylaxis in D614G infected K18-hACE2 mice. Animals received intranasally (i.n.) 5 mg/kg VHH-B07-Fc (B07-Fc) or 5 mg/kg VHH-Fc ctl (ctl-Fc). Twenty-four hours later, they were infected with 10 4 PFU D614G SARS-CoV-2 i.n. Mice were clinically monitored between days 3 to 6. Mice were euthanized on day 6 and lungs were collected for analysis. g Body weight measured over time post-infection in ctl-Fc ( n = 5) and B07-Fc-treated groups. Mice of the B07-Fc group were split into VHH_high ( n = 4) and VHH_low ( n = 6) according to the B07-Fc concentration measured in the lung on day 6 p.i. (higher or lower than 200 ng/g of lung, respectively). Data are presented as mean values +/- SD. Linear mixed model two sided (no adjustment for multiple comparisons): ns, nonsignificant (p = 0.26); ** p = 0.00378; *** p = 0.000396. h Relationship between body weight loss and B07-Fc concentration quantified in the lung by ELISA on day 6 p.i. Source Data are provided as a Source Data file.

Techniques: Infection, Quantitative RT-PCR, Staining, Immunofluorescence, Concentration Assay, Enzyme-linked Immunosorbent Assay

( A ) Schematic showing whole-genome CRISPRa screen in HEK293 -ACE2 cells using the Weissman library ( https://weissman.wi.mit.edu/crispr/ ). Cells were transduced with lentiviral pools encoding individual activation sgRNAs tiling the genome. Cells were then inoculated with authentic SARS-CoV-2 or parallel controls, and guides promoting SARS-CoV-2 were identified by sequencing. ( N = 2.) ( B ) Top genes by local FDR (locFDR) (<0.4 considered significant) identified after selection. See also . ( C ) Plot showing sgRNA enrichment (log fold change) versus gene ranking. ( D and E ) Top pathways identified using Ingenuity Pathway Analysis (IPA) ( D ) or KEGG pathways ( E ). ( F and G ) Independent validation of top hits including cytosolic ( ARL4C , LRRC29 , RPS16 ) or mitochondrial proteins (MRPS7) ( F ), and membrane proteins promoting SARS-CoV-2 resistance (48 hours after inoculation) ( G ). Significance was determined by 2-way ANOVA and Dunnett’s test. ( N = 3.) ( H ) Diagram of SARS-CoV-2 FACS spike binding assay. ( I ) CRISPRa-driven expression of SELP/P selectin promotes binding to SARS-CoV-2 spike protein. Significance was obtained by 1-way ANOVA and Šidák’s test, * P < 0.05. ( N = 3.)

Journal: The Journal of Clinical Investigation

Article Title: P selectin promotes SARS-CoV-2 interactions with platelets and the endothelium

doi: 10.1172/JCI184514

Figure Lengend Snippet: ( A ) Schematic showing whole-genome CRISPRa screen in HEK293 -ACE2 cells using the Weissman library ( https://weissman.wi.mit.edu/crispr/ ). Cells were transduced with lentiviral pools encoding individual activation sgRNAs tiling the genome. Cells were then inoculated with authentic SARS-CoV-2 or parallel controls, and guides promoting SARS-CoV-2 were identified by sequencing. ( N = 2.) ( B ) Top genes by local FDR (locFDR) (<0.4 considered significant) identified after selection. See also . ( C ) Plot showing sgRNA enrichment (log fold change) versus gene ranking. ( D and E ) Top pathways identified using Ingenuity Pathway Analysis (IPA) ( D ) or KEGG pathways ( E ). ( F and G ) Independent validation of top hits including cytosolic ( ARL4C , LRRC29 , RPS16 ) or mitochondrial proteins (MRPS7) ( F ), and membrane proteins promoting SARS-CoV-2 resistance (48 hours after inoculation) ( G ). Significance was determined by 2-way ANOVA and Dunnett’s test. ( N = 3.) ( H ) Diagram of SARS-CoV-2 FACS spike binding assay. ( I ) CRISPRa-driven expression of SELP/P selectin promotes binding to SARS-CoV-2 spike protein. Significance was obtained by 1-way ANOVA and Šidák’s test, * P < 0.05. ( N = 3.)

Article Snippet: Transgenic mice expressing human ACE2 (hACE2) were purchased from Taconic Biosciences [B6;C3-Tg(CAG-ACE2)70Ctkt] and then bred in-house.

Techniques: CRISPR, Transduction, Activation Assay, Sequencing, Selection, Biomarker Discovery, Membrane, Binding Assay, Expressing

( A ) Ectopic ACE2 or P selectin expression strategy. Scale bars: 60 µm. ( B ) Representative flow cytometry plots showing binding of P selectin to SARS-CoV-2 spike protein. ( C ) Quantification of B . Significance was determined by paired 2-tailed t test, ** P < 0.01. ( N = 3.) ( D ) Schematic of DiD-labeled pseudolentivirus assay used in E and F . ( E and F ) Flow cytometry histograms ( E ) and quantification of binding intensity ( F ) presented for pseudovirus expressing MERS, SARS-CoV-1, and SARS-CoV-2 A2.2 and Delta variants. MFI, mean fluorescence intensity. Significance was determined by 1-way ANOVA and Dunnett’s test, * P < 0.05, ** P < 0.01. ( N = 3.) ( G ) Binding sites (red) for blocking antibodies AZD1061 (cilgavimab), AZD8895 (tixagevimab), and S309 (sotrovimab) and predicted binding site for P selectin (blue) on SARS-CoV-2 spike glycoprotein (Protein Data Bank: 7WK2) . ( H ) Key predicted interactive regions between P selectin and spike RBD. C-type lectin-like domain, CTLD. ( I ) Binding of antibody-blocked spike protein in HEK293T cells expressing P selectin. Significance was determined by 1-way ANOVA and Dunnett’s test, ** P < 0.01, *** P < 0.001. ( N = 3).

Journal: The Journal of Clinical Investigation

Article Title: P selectin promotes SARS-CoV-2 interactions with platelets and the endothelium

doi: 10.1172/JCI184514

Figure Lengend Snippet: ( A ) Ectopic ACE2 or P selectin expression strategy. Scale bars: 60 µm. ( B ) Representative flow cytometry plots showing binding of P selectin to SARS-CoV-2 spike protein. ( C ) Quantification of B . Significance was determined by paired 2-tailed t test, ** P < 0.01. ( N = 3.) ( D ) Schematic of DiD-labeled pseudolentivirus assay used in E and F . ( E and F ) Flow cytometry histograms ( E ) and quantification of binding intensity ( F ) presented for pseudovirus expressing MERS, SARS-CoV-1, and SARS-CoV-2 A2.2 and Delta variants. MFI, mean fluorescence intensity. Significance was determined by 1-way ANOVA and Dunnett’s test, * P < 0.05, ** P < 0.01. ( N = 3.) ( G ) Binding sites (red) for blocking antibodies AZD1061 (cilgavimab), AZD8895 (tixagevimab), and S309 (sotrovimab) and predicted binding site for P selectin (blue) on SARS-CoV-2 spike glycoprotein (Protein Data Bank: 7WK2) . ( H ) Key predicted interactive regions between P selectin and spike RBD. C-type lectin-like domain, CTLD. ( I ) Binding of antibody-blocked spike protein in HEK293T cells expressing P selectin. Significance was determined by 1-way ANOVA and Dunnett’s test, ** P < 0.01, *** P < 0.001. ( N = 3).

Article Snippet: Transgenic mice expressing human ACE2 (hACE2) were purchased from Taconic Biosciences [B6;C3-Tg(CAG-ACE2)70Ctkt] and then bred in-house.

Techniques: Expressing, Flow Cytometry, Binding Assay, Labeling, Fluorescence, Blocking Assay

( A ) Images of ACE2- or P selectin–expressing HEK293T cells infected with SARS-CoV-2 spike-expressing pseudolentivirus. Scale bars: 60 μm. ( B ) Quantification of A . Significance was determined by 2-way ANOVA and Dunnett’s test, **** P < 0.001. ( N = 3.) ( C and D ) Infection of HEK293-ACE2/TMPRSS2–expressing cells with SARS-CoV-2 A2.2 ( C ) or Delta ( D ) variant is suppressed by coexpression of SELP cDNA. Significance was determined by 2-way ANOVA and Šidák’s test. * P < 0.05, ** P < 0.01, *** P < 0.001. ( N = 3). ( E ) Plaque assay showing plaque-forming units (PFU) in VeroE6/TMPRSS2 cells. Significance was determined by paired t test, * P < 0.05. ( N = 4.) ( F ) Infection of HEK293-ACE2/TMPRSS2–expressing cells with SARS-CoV-2 A2.2 is suppressed by SELP mRNA expression. Significance was determined by 2-way ANOVA and Šidák’s test, * P < 0.05, ** P < 0.01, *** P < 0.001. ( N = 3.) ( G ) Schematic showing infection of mice with a lethal dose of SARS-CoV-2 and daily injection of P selectin blocking antibody to evaluate infection course. ( H ) Body weight changes after lethal SARS-CoV-2 inoculations in the presence of P selectin blocking antibody. Significance was determined by 2-way ANOVA and Šidák’s test, * P < 0.05. ( N = 8–9.)

Journal: The Journal of Clinical Investigation

Article Title: P selectin promotes SARS-CoV-2 interactions with platelets and the endothelium

doi: 10.1172/JCI184514

Figure Lengend Snippet: ( A ) Images of ACE2- or P selectin–expressing HEK293T cells infected with SARS-CoV-2 spike-expressing pseudolentivirus. Scale bars: 60 μm. ( B ) Quantification of A . Significance was determined by 2-way ANOVA and Dunnett’s test, **** P < 0.001. ( N = 3.) ( C and D ) Infection of HEK293-ACE2/TMPRSS2–expressing cells with SARS-CoV-2 A2.2 ( C ) or Delta ( D ) variant is suppressed by coexpression of SELP cDNA. Significance was determined by 2-way ANOVA and Šidák’s test. * P < 0.05, ** P < 0.01, *** P < 0.001. ( N = 3). ( E ) Plaque assay showing plaque-forming units (PFU) in VeroE6/TMPRSS2 cells. Significance was determined by paired t test, * P < 0.05. ( N = 4.) ( F ) Infection of HEK293-ACE2/TMPRSS2–expressing cells with SARS-CoV-2 A2.2 is suppressed by SELP mRNA expression. Significance was determined by 2-way ANOVA and Šidák’s test, * P < 0.05, ** P < 0.01, *** P < 0.001. ( N = 3.) ( G ) Schematic showing infection of mice with a lethal dose of SARS-CoV-2 and daily injection of P selectin blocking antibody to evaluate infection course. ( H ) Body weight changes after lethal SARS-CoV-2 inoculations in the presence of P selectin blocking antibody. Significance was determined by 2-way ANOVA and Šidák’s test, * P < 0.05. ( N = 8–9.)

Article Snippet: Transgenic mice expressing human ACE2 (hACE2) were purchased from Taconic Biosciences [B6;C3-Tg(CAG-ACE2)70Ctkt] and then bred in-house.

Techniques: Expressing, Infection, Variant Assay, Plaque Assay, Injection, Blocking Assay

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