his rbd jn 1  (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: 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: bioRxiv

Article Title: Heterogeneous hybrid immunity against Omicron variant JN.1 at 11 months following breakthrough infection

doi: 10.1101/2024.03.02.583082

Figure Lengend Snippet: ( A ) The frequency of each variant is calculated as a centered 14-day rolling average. ( B ) The most representative amino acid mutations of each variant were selected to construct pseudotyped virus for this study. Each mutation in included SARS-CoV-2 variant is indicated relative to the reference D614G sequence. A dot indicates an identical amino acid at the indicated position, while a dash indicates a deletion at that point. NTD N-terminal domain, RBD Receptor-binding domain, FP Fusion peptide, HR1 Heptad repeat 1, HR2 Heptad repeat 2, TM transmembrane domain, CTD C-terminal domain.

Article Snippet: The plates were incubated with biotinylated SARS-CoV-2 (JN.1) spike RBD protein (Sino Biological, 1 μg/ml) for 2 h and Streptavidin-AP Conjugate from Invitrogen (1.5 μg/ml) for 1 h, followed by the addition of NBT/BCIP chromogen substrate solution for 12 min.

Techniques: Variant Assay, Construct, Virus, Mutagenesis, Sequencing, Binding Assay

Journal: bioRxiv

Article Title: Heterogeneous hybrid immunity against Omicron variant JN.1 at 11 months following breakthrough infection

doi: 10.1101/2024.03.02.583082

Figure Lengend Snippet: Comparison of SARS-CoV-2 Spike RBD IgG titers ( A ) and SARS-CoV-2 pseduovirus neutralizing antibody titers ( B ) in response to prototype strain and Omicron lineages including BA.5, XBB, EG.5.1 and JN.1 in blood from subjects grouped as: unvaccinated children with BTI (n=4); vaccinated children with BTI (n=10); vaccinated adolescents with BTI (n=14); vaccinated adults with BTI (n=14); vaccinated adults with BTI (n=30); vaccinated seniors with BTI (n=30); vaccinated adults with BTI plus RI (n=13); vaccinated seniors with BTI plus RI (n=8). Dotted lines reflect lower limits of quantitation. Bold horizontal lines and numbers above reflect median values.

Article Snippet: The plates were incubated with biotinylated SARS-CoV-2 (JN.1) spike RBD protein (Sino Biological, 1 μg/ml) for 2 h and Streptavidin-AP Conjugate from Invitrogen (1.5 μg/ml) for 1 h, followed by the addition of NBT/BCIP chromogen substrate solution for 12 min.

Techniques: Comparison, Quantitation Assay

Journal: bioRxiv

Article Title: Heterogeneous hybrid immunity against Omicron variant JN.1 at 11 months following breakthrough infection

doi: 10.1101/2024.03.02.583082

Figure Lengend Snippet: The Antigenic maps were generated based on binding antibody data ( A ), and neutralizing antibody data ( B ) for all cohorts. The SARS-CoV-2 variants are shown as circles and labeled. Sera sample are shown as empty squares. Both axes represent the antigenic unit (AU) corresponding to fold change in the antibody titers.

Article Snippet: The plates were incubated with biotinylated SARS-CoV-2 (JN.1) spike RBD protein (Sino Biological, 1 μg/ml) for 2 h and Streptavidin-AP Conjugate from Invitrogen (1.5 μg/ml) for 1 h, followed by the addition of NBT/BCIP chromogen substrate solution for 12 min.

Techniques: Generated, Binding Assay, Labeling

Journal: bioRxiv

Article Title: Heterogeneous hybrid immunity against Omicron variant JN.1 at 11 months following breakthrough infection

doi: 10.1101/2024.03.02.583082

Figure Lengend Snippet: Paired-sample comparison of NAb titers against prototype strain and Omicron lineages including BA.5, XBB, EG.5.1 and JN.1 was conducted for blood (blue) and BAL (red) (n=17).

Article Snippet: The plates were incubated with biotinylated SARS-CoV-2 (JN.1) spike RBD protein (Sino Biological, 1 μg/ml) for 2 h and Streptavidin-AP Conjugate from Invitrogen (1.5 μg/ml) for 1 h, followed by the addition of NBT/BCIP chromogen substrate solution for 12 min.

Techniques: Comparison

Journal: bioRxiv

Article Title: Heterogeneous hybrid immunity against Omicron variant JN.1 at 11 months following breakthrough infection

doi: 10.1101/2024.03.02.583082

Figure Lengend Snippet: NAb titers against prototype, BA.5, XBB, EG.5.1 and JN.1 in BAL from vaccinated subjects with BTI (orange; n=30). or without BTI (grey; n=7). Dotted lines reflect lower limits of quantitation.

Article Snippet: The plates were incubated with biotinylated SARS-CoV-2 (JN.1) spike RBD protein (Sino Biological, 1 μg/ml) for 2 h and Streptavidin-AP Conjugate from Invitrogen (1.5 μg/ml) for 1 h, followed by the addition of NBT/BCIP chromogen substrate solution for 12 min.

Techniques: Quantitation Assay

Journal: bioRxiv

Article Title: Heterogeneous hybrid immunity against Omicron variant JN.1 at 11 months following breakthrough infection

doi: 10.1101/2024.03.02.583082

Figure Lengend Snippet: ( A ) Circulating memory B cells responding to prototype and JN.1 variants were measured in blood from BTI groups with RI (n=12) or without RI (n=11). ( B ) Representative flow cytometry plot show expression of CD21 and CD27 on prototype (blue) and JN.1(orange) RBD-specific memory B cells, gated on IgD-B cells in blood from subject with BTI. The phenotype of activated memory (AM; red) and resting memory (RM; grey) was indicated for circulating memory B cells responding to prototype and JN.1. ( C ) Circulating memory B cells responding to prototype and JN.1 variants were measured in BAL from BTI groups (n=12). ( D ) Representative flow cytometry plot show expression of CD21 and CD27 on prototype (blue) and JN.1(orange) RBD-specific memory B cells, gated on IgD-B cells in BAL from subject with BTI. The phenotype of AM (red) and RM (grey) was indicated for lung-resident memory B cells responding to prototype and JN.1. Box and whisker plots reflect interquartile ranges (boxes), medians (horizontal lines), and range (whiskers).

Article Snippet: The plates were incubated with biotinylated SARS-CoV-2 (JN.1) spike RBD protein (Sino Biological, 1 μg/ml) for 2 h and Streptavidin-AP Conjugate from Invitrogen (1.5 μg/ml) for 1 h, followed by the addition of NBT/BCIP chromogen substrate solution for 12 min.

Techniques: Flow Cytometry, Expressing, Whisker Assay

Journal: bioRxiv

Article Title: Heterogeneous hybrid immunity against Omicron variant JN.1 at 11 months following breakthrough infection

doi: 10.1101/2024.03.02.583082

Figure Lengend Snippet: ( A ) JN.1 RBD-binding IgG-secreting memory B cells were detected in PBMCs from BTI subjects with binding antibody (Ab) titers >500 (red) or <500 (blue). Dotted line reflects lower limits of quantitation. ( B ) Representative ELISpot wells coated with JN.1 RBD and developed after plating the stimulated PBMCs from BTI subjects with binding antibody titers >500 (above) or titers <500 (below). ( C ) Correlation of JN.1 RBD-specific memory B cell ELISpot data with ELISA binding antibody titers in BTI subjects. Bold line indicates best linear fits. P and R values indicate two-sided Spearman rank-correlation tests.

Article Snippet: The plates were incubated with biotinylated SARS-CoV-2 (JN.1) spike RBD protein (Sino Biological, 1 μg/ml) for 2 h and Streptavidin-AP Conjugate from Invitrogen (1.5 μg/ml) for 1 h, followed by the addition of NBT/BCIP chromogen substrate solution for 12 min.

Techniques: Binding Assay, Quantitation Assay, Enzyme-linked Immunospot, Enzyme-linked Immunosorbent Assay