Journal: PLOS Pathogens

Article Title: Determinants of natural killer cell-mediated antibody dependent cellular cytotoxicity in SARS-CoV-2 antibodies

doi: 10.1371/journal.ppat.1014026

Figure Lengend Snippet: (A) Distribution of C68 Abs across SARS-CoV-2 Spike protein domains. Pie chart segments indicate total Abs targeting each domain: NTD (22), RBD (69), S2 (45), and CTD (6). (B) Schematic of full-length Spike and cryo-EM structure of the WH-1 SARS-CoV-2 Spike trimer (PDB: 6VSB), with domains color-coded to match the pie chart and bar graph designations NTD (blue), RBD (green), CTD (cyan), S2 (magenta); adapted from . (C) Proportion of ADCC-capable Abs across domains of the WH-1 SARS-CoV-2 spike protein. The numerical fractions above each bar indicate the number of ADCC capable Abs relative to the total number of Abs against the indicated domain. Abs were defined as ADCC-capable if the average %CD107a ⁺ NK cells from two independent experiments exceeded 5%, as described in methods. Pairwise comparisons were performed using Fisher exact test with Holm correction for multiple testing. Of note, CTD Abs are clonally restricted, limiting our ability to draw conclusions. (D) ADCC potency of Abs by Spike domain. Each point represents a single Ab. Boxplots show median, IQR, and 1.5 × IQR whiskers. Dotted line indicates limit of detection. Wilcoxon tests with Holm correction; significance indicated as *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; ns = not significant. (E) Proportion of Abs that neutralize pseudovirus bearing the WH-1 SARS-CoV-2 Spike protein shown by epitope domain target. The numerical fractions above each bar indicate the number of neutralizing Abs relative to the total number of Abs against the indicated domain. Neutralization status was considered positive when IC₅₀ < 20,000 ng/mL as described in methods. Pairwise comparisons were performed using Fisher exact test with Holm correction for multiple testing. F) Neutralization potency (log₁₀[IC₅₀, ng/mL]) by Spike domain. Dotted line indicates limit of detection. Wilcoxon tests with Holm correction as in (D).

Article Snippet: The proteins used included: SARS-CoV-2 S1 WH-1 strain (Sino Biological, cat. 40591-V08H), SARS-CoV-2 S2 (mammalian cell-derived, cat. 40590-V08H1), SARS-CoV-2 NTD subunit (Sino Biologicals, 40591-V49H) and SARS-CoV-2 RBD (Sino Biological, cat. 40592-V08H).

Techniques: Cryo-EM Sample Prep, Neutralization

Journal: PLOS Pathogens

Article Title: Determinants of natural killer cell-mediated antibody dependent cellular cytotoxicity in SARS-CoV-2 antibodies

doi: 10.1371/journal.ppat.1014026

Figure Lengend Snippet: Correlation between binding to WH-1 SARS-CoV-2 Spike (MFI) and ADCC potency (% CD107a + NK cells) for binder Abs (MFI > 1000). Each data point represents individual Abs. Pearson correlation coefficient (R), p value and 95% CI (gray) are shown.

Article Snippet: The proteins used included: SARS-CoV-2 S1 WH-1 strain (Sino Biological, cat. 40591-V08H), SARS-CoV-2 S2 (mammalian cell-derived, cat. 40590-V08H1), SARS-CoV-2 NTD subunit (Sino Biologicals, 40591-V49H) and SARS-CoV-2 RBD (Sino Biological, cat. 40592-V08H).

Techniques: Binding Assay

Journal: PLOS Pathogens

Article Title: Determinants of natural killer cell-mediated antibody dependent cellular cytotoxicity in SARS-CoV-2 antibodies

doi: 10.1371/journal.ppat.1014026

Figure Lengend Snippet: (A) Pie charts showing the distribution of C68 mAb across SARS CoV-2 Spike domain (NTD, RBD, S2, CTD) at post-vaccination infection 1 (PVI-1) and PVI-2. (B–C) Boxplots comparing (B) neutralization potency (log₁₀ IC50 in ng/mL), and (C) ADCC activity between Abs isolated at PVI-1 and PVI-2. Each dot represents one mAb. Dotted line indicates limit of detection. Statistical comparisons were performed using the Wilcoxon rank-sum test; p-values are shown above each plot.

Article Snippet: The proteins used included: SARS-CoV-2 S1 WH-1 strain (Sino Biological, cat. 40591-V08H), SARS-CoV-2 S2 (mammalian cell-derived, cat. 40590-V08H1), SARS-CoV-2 NTD subunit (Sino Biologicals, 40591-V49H) and SARS-CoV-2 RBD (Sino Biological, cat. 40592-V08H).

Techniques: Infection, Neutralization, Activity Assay, Isolation

Journal: PLOS Pathogens

Article Title: Determinants of natural killer cell-mediated antibody dependent cellular cytotoxicity in SARS-CoV-2 antibodies

doi: 10.1371/journal.ppat.1014026

Figure Lengend Snippet: Scatter plots showing the correlation between CDR3 length (VH (A), VL (B) and ADCC activity against WH-1 SARS-CoV-2. Analysis includes ADCC capable Abs. Pearson correlation coefficient (R), p-value, and 95% CI (gray shading) are shown.

Article Snippet: The proteins used included: SARS-CoV-2 S1 WH-1 strain (Sino Biological, cat. 40591-V08H), SARS-CoV-2 S2 (mammalian cell-derived, cat. 40590-V08H1), SARS-CoV-2 NTD subunit (Sino Biologicals, 40591-V49H) and SARS-CoV-2 RBD (Sino Biological, cat. 40592-V08H).

Techniques: Activity Assay

Journal: Communications Biology

Article Title: Endemic penetrance of SARS-CoV-2 has impacted marginally on immunity to spike protein of human coronaviruses

doi: 10.1038/s42003-025-09474-x

Figure Lengend Snippet: Spike-specific IgG titre against SARS-CoV-2, OC43, HKU1, NL63 and 229E in SARS-CoV-2-seronegative unvaccinated donors ( a , n = 124), SARS-CoV-2-seropositive unvaccinated donors ( b , n = 152), SARS-CoV-2-seronegative vaccinated donors ( c , n = 24) and SARS-CoV-2-seropositive vaccinated donors ( d , n = 69). Kruskal–Wallis (Dunn’s multiple comparisons test). Black lines indicate median titre and interquartile range, dotted black line indicates the cutoff. Correlation of age with Spike-specific IgG titre against SARS-CoV-2 (black), OC43 (blue) and HKU1 (green) in seronegative unvaccinated donors ( e , n = 124), seropositive unvaccinated donors ( f , n = 152), seronegative vaccinated donors ( g , n = 24) and seropositive vaccinated donors ( h , n = 69). The Spearman correlation coefficient ( r ) and P -values are shown. Fitted lines are linear regressions. No age-related correlations were observed with HCoVs (Supplementary Fig. ). i Fold change in median SARS-CoV-2 spike IgG titre in seropositive unvaccinated (light red), seronegative vaccinated (black) and seropositive vaccinated (red) donors in relation to seronegative unvaccinated donors. Fold changes are listed. j Fold change in median OC43, HKU1, NL63 and 229E spike-specific IgG in seropositive unvaccinated (light red), seronegative vaccinated (black) and seropositive vaccinated (red) donors in relation to seronegative unvaccinated donors (grey). Fold changes are listed, black dotted line denotes a fold change of 1. Source data: Supplementary Data .

Article Snippet: Donor plasma was diluted 1:10 in PBS prior to incubation at 37 °C for 30 min with an equal volume of either recombinant SARS-CoV-2 S1 protein (R&D Systems, 10569-CV-100) or recombinant SARS-CoV-2 S2 protein (R&D Systems, 10594-CV-100) at a concentration of 500 μg/ml in PBS, or with PBS alone.

Techniques:

Journal: Communications Biology

Article Title: Endemic penetrance of SARS-CoV-2 has impacted marginally on immunity to spike protein of human coronaviruses

doi: 10.1038/s42003-025-09474-x

Figure Lengend Snippet: Correlation of SARS-CoV-2 spike IgG titre with alphacoronavirus spike (NL63 (green) and 229E (yellow), a – d ) and betacoronavirus spike (OC43 (purple) and HKU1 (pink), e – h ) IgG titres in seronegative unvaccinated donors ( a , n = 124), SARS-CoV-2-seropositive unvaccinated donors ( b , n = 152), SARS-CoV-2-seronegative vaccinated donors ( c , n = 24) and SARS-CoV-2-seropositive vaccinated donors ( d n = 69). The Spearman correlation coefficient ( r ) and P -values are shown. Source data: Supplementary Data .

Article Snippet: Donor plasma was diluted 1:10 in PBS prior to incubation at 37 °C for 30 min with an equal volume of either recombinant SARS-CoV-2 S1 protein (R&D Systems, 10569-CV-100) or recombinant SARS-CoV-2 S2 protein (R&D Systems, 10594-CV-100) at a concentration of 500 μg/ml in PBS, or with PBS alone.

Techniques:

Journal: Communications Biology

Article Title: Endemic penetrance of SARS-CoV-2 has impacted marginally on immunity to spike protein of human coronaviruses

doi: 10.1038/s42003-025-09474-x

Figure Lengend Snippet: SARS-CoV-2 ( a ), OC43 ( b ), HKU1 ( c ), NL63 ( d ) and 229E ( e ) spike-specific IgG binding from unvaccinated (pink, n = 18) or vaccinated (orange, n = 23) SARS-CoV-2-seropositive donors following pre-adsorption with either S1 or S2 domains from SARS-CoV-2 Spike protein. Data are expressed as a percentage compared to binding within the unadsorbed control (Control). Friedman test with Dunn’s multiple comparison test was used. Source data: Supplementary Data .

Article Snippet: Donor plasma was diluted 1:10 in PBS prior to incubation at 37 °C for 30 min with an equal volume of either recombinant SARS-CoV-2 S1 protein (R&D Systems, 10569-CV-100) or recombinant SARS-CoV-2 S2 protein (R&D Systems, 10594-CV-100) at a concentration of 500 μg/ml in PBS, or with PBS alone.

Techniques: Binding Assay, Adsorption, Control, Comparison

Journal: Communications Biology

Article Title: Endemic penetrance of SARS-CoV-2 has impacted marginally on immunity to spike protein of human coronaviruses

doi: 10.1038/s42003-025-09474-x

Figure Lengend Snippet: a IFNγ FluoroSpot response to SARS-CoV-2 and HCoV S1 peptide stimulation ( n = 66–87). Kruskal–Wallis (Dunn’s multiple comparisons test). Black lines indicate the median and interquartile range. b IFNγ FluoroSpot response to SARS-CoV-2 and HCoV S2 peptide stimulation ( n = 64–86). Kruskal–Wallis (Dunn’s multiple comparisons test). Black lines indicate the median and interquartile range. c Ratio of S2-specific to S1-specific IFNγ cellular responses in SARS-CoV-2 and HCoV ( n = 54–84). Betacoronaviruses are indicated with purple dots, and alphacoronaviruses are indicated with blue dots. Kruskal–Wallis (Dunn’s multiple comparisons test). Black lines indicate the median and interquartile range. d Ratio of S2-specific to S1-specific IFNγ cellular responses to SARS-CoV-2 (black dots, n = 84), alphacoronaviruses NL63 and 229E (blue dots, n = 66) and betacoronaviruses OC43 and HKU1 (purple dots, n = 74) in relation to age. Dotted black line indicates a S2:S1 ratio of 1. The Spearman correlation coefficient and P -values are shown. Fitted lines are linear regressions. Source data: Supplementary Data .

Article Snippet: Donor plasma was diluted 1:10 in PBS prior to incubation at 37 °C for 30 min with an equal volume of either recombinant SARS-CoV-2 S1 protein (R&D Systems, 10569-CV-100) or recombinant SARS-CoV-2 S2 protein (R&D Systems, 10594-CV-100) at a concentration of 500 μg/ml in PBS, or with PBS alone.

Techniques:

Journal: Communications Biology

Article Title: Endemic penetrance of SARS-CoV-2 has impacted marginally on immunity to spike protein of human coronaviruses

doi: 10.1038/s42003-025-09474-x

Figure Lengend Snippet: a Mean fluorescence intensity (MFI) of CXCL8 within FluoroSpot eluates after stimulation with either S1 or S2 spike peptides from SARS-CoV-2 or OC43 and HKU1 compared to DMSO control. Donors were SARS-CoV-2 vaccinated donors and >65 years of age ( n = 15). Kruskal–Wallis (uncorrected Dunn’s test). Black lines indicate the median MFI with interquartile range. b Mean fluorescence intensity (MFI) of CXCL10, TNFɑ, IFN-ɑ2, IL-7 and IL-10 within FluoroSpot eluates after stimulation with SARS-CoV-2 S1 or S2 peptides compared to a DMSO control in vaccinated donors over the age of 65 ( n = 15). Kruskal–Wallis (uncorrected Dunn’s test). Black lines indicate the median MFI with interquartile range. Source data: Supplementary Data .

Article Snippet: Donor plasma was diluted 1:10 in PBS prior to incubation at 37 °C for 30 min with an equal volume of either recombinant SARS-CoV-2 S1 protein (R&D Systems, 10569-CV-100) or recombinant SARS-CoV-2 S2 protein (R&D Systems, 10594-CV-100) at a concentration of 500 μg/ml in PBS, or with PBS alone.

Techniques: Fluorescence, Control