Journal: bioRxiv

Article Title: HIV broadly neutralizing antibodies with enhanced Fc-CD16 affinity increase NK cell ADCC even with limited envelope binding

doi: 10.64898/2026.04.27.721006

Figure Lengend Snippet: Cytokine stimulation enhances NK cell activation while downregulating CD16 expression through ADAM17-dependent shedding. ( A ) Representative flow plots of CD69 induction of NK cells stimulated with IL-15, IL-12/IL-15, or IL-12/IL-15/IL-18. ( B ) CD69 induction across donors (n=10). ( C ) Viability upon different treatments. ( D ) Changes in CD16 expression across multiple donors upon cytokine treatment. ( E ) Spearman correlation analysis between CD16 expression and CD69 induction. ( F ) Representative histogram of CD16 expression upon triple cytokine treatment in the presence of TAPI-I (right panel) or DMSO control (left panel). ( G ) CD16 expression in donors (n=13) stimulated with cytokines in the absence or presence of TAPI-1. ( H ) Effects of TAPI-I on NK cell activation. ( I ) Percentage remaining p24 + cells upon coculture of HIV-infected CD4 + T cells with autologous NK cells from donors (n=7) pre-stimulated with cytokines in the presence of TAPI-1 or DMSO control. ( J ) Percentage ADCC calculated as indicated in methods from data shown in ( I ). Statistical significance was calculated using one-way ANOVA with multiple comparison compared to the no cytokine condition for panel ( B-D ), two-way ANOVA with multiple comparison compared to the IL12/IL15/IL18 condition for panel ( I ), and two-way ANOVA for panel ( J ). Open symbols represent female donors and closed symbols male donors.

Article Snippet: Ten cytokines were measured using Quanterix SP-X Corplex Cytokine Panel (IFN-γ, IL-1β, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, IL-22, TNF-α) (cat# 85-0329).

Techniques: Activation Assay, Expressing, Control, Infection, Comparison

Journal: bioRxiv

Article Title: HIV broadly neutralizing antibodies with enhanced Fc-CD16 affinity increase NK cell ADCC even with limited envelope binding

doi: 10.64898/2026.04.27.721006

Figure Lengend Snippet: Functional activity, cytokine secretion, and Fc binding of VRC07-523-LS and Fc variants. ( A ) Table summarizing VRC07-523LS antibody mutants with optimized Fc biding including specific mutations and effects on CD16 binding. ( B ) CD16 reporter activity in Jurkat-Lucia NFAT-CD16 cells exposed to antibody variants. ( C ) CD69 induction across donors (n=7). ( D ) Viability upon different conditions. ( E-O ) Supernatants from NK cells cultured overnight with WT VRC07-523-LS or Fc mutant antibodies were analyzed for cytokine secretion (fg/mL). Levels of ( E ) IFN-γ (Limit of detection (LOD) 1fg/mL), ( F ) TNF-α (LOD 16.7fg/mL), ( G ) IL-1β (LOD 16.3fg/mL), ( H ) IL-4 (LOD 339.1fg/mL), ( I ) IL-5 (LOD 8.7fg/ml), ( J ) IL-6 (LOD 21.1fg/mL), ( K ) IL-8 (LOD 41.7fg/mL), ( M ) IL-10 (LOD 16.8fg/mL), ( N ) IL-12p70 (LOD 24.7 fg/mL), ( O ) IL-22 (LOD 4.5 fg/mL) (n=6). ( P ) Representative flow cytometry histograms showing VRC07-523LS binding to surface Env on uninfected (blue), uninfected exposed (red), or HIV-infected cells (black). A FMO control is included (grey). Quantification of Env binding for the different antibodies (n=7). ( Q ) Surface expression of CD107a upon co-culture of NK cells alone (no CD4 + ), or NK cells co-cultured with uninfected autologous CD4 + or NL4-3 infected CD4 + T cells in the presence of the indicated antibody (n=2). Statistical significance was assessed using a Friedman test with multiple comparisons compared to unstimulated condition for panels ( C-O ), and two-way ANOVA with multiple comparisons for panel ( P ). Open symbols represent female and closed symbols male donors.

Article Snippet: Ten cytokines were measured using Quanterix SP-X Corplex Cytokine Panel (IFN-γ, IL-1β, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, IL-22, TNF-α) (cat# 85-0329).

Techniques: Functional Assay, Activity Assay, Binding Assay, Cell Culture, Mutagenesis, Flow Cytometry, Infection, Control, Expressing, Co-Culture Assay

Journal: bioRxiv

Article Title: HIV broadly neutralizing antibodies with enhanced Fc-CD16 affinity increase NK cell ADCC even with limited envelope binding

doi: 10.64898/2026.04.27.721006

Figure Lengend Snippet: Enhancing antibody affinity to CD16 improves NK cell-mediated ADCC against HIV-infected cells independently of cytokine activation or ADAM17 inhibition. Percent of remaining HIV p24⁺ infected CD4⁺ T cells following co-culture with autologous NK cells from donors pre-treated with or without cytokines (IL-12/IL-15/IL-18) (n=16) ( A ) and ADAM17 inhibitor TAPI-1 (n=7) ( B ), in the presence of the indicated antibody. ( C ) Hierarchical cluster analysis of percentage of remaining p24⁺ cells in different conditions shown in A-B. ADCC activity under the same treatment conditions in the absence ( D ) or the presence ( E ) of the ADAM17 inhibitor TAPI-1. Statistical significance was determined using two-way ANOVA with multiple comparison compared to WT. Open symbols represent female and closed symbols male donors. ( F ) Hierarchical cluster analysis of ADCC in different conditions shown in D-E . Clusters were done using ClustVis.

Article Snippet: Ten cytokines were measured using Quanterix SP-X Corplex Cytokine Panel (IFN-γ, IL-1β, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, IL-22, TNF-α) (cat# 85-0329).

Techniques: Infection, Activation Assay, Inhibition, Co-Culture Assay, Activity Assay, Comparison

Journal: bioRxiv

Article Title: HIV broadly neutralizing antibodies with enhanced Fc-CD16 affinity increase NK cell ADCC even with limited envelope binding

doi: 10.64898/2026.04.27.721006

Figure Lengend Snippet: Effects of Fc-optimized VRC07-523LS mutants across HIV-1 subtypes. ( A ) Neutralization curves of VRC07-523LS against indicated HIV-1 viral strain. ( B ) Env binding of the different antibodies to HIV-infected CD4T cells with the indicated viral strain (n=3-4). ( C ) Spearman correlation between Env binding and neutralization IC 50 values. ( D ) Percentage of remaining p24 + cells following natural cytotoxicity by autologous NK cells pretreated with a combination of cytokines or no cytokine control (n= 6-16). ( E-I ) Percentage of remaining p24+ cells for each of the indicated viruses in the presence of the different antibodies (n= 7-8). Statistical significance was assessed using a one-sample t test with a hypothetical value set to 1 for panel ( B ), wilcoxon signed rank test with hypothetical value set to 100 for panel ( E ), and two -way ANOVA with multiple comparison compared to WT for panels ( E–I ). Open symbols represent female and closed symbols male donors. ( J ) Hierarchical cluster analysis of the percentage remaining p24 + cells. Clusters were done using ClustVis.

Article Snippet: Ten cytokines were measured using Quanterix SP-X Corplex Cytokine Panel (IFN-γ, IL-1β, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, IL-22, TNF-α) (cat# 85-0329).

Techniques: Neutralization, Binding Assay, Infection, Control, Comparison

Journal: bioRxiv

Article Title: HIV broadly neutralizing antibodies with enhanced Fc-CD16 affinity increase NK cell ADCC even with limited envelope binding

doi: 10.64898/2026.04.27.721006

Figure Lengend Snippet: Cytokine activation of NK cells requires higher levels of surface Env binding to promote ADCC. ( A ) Percentage of ADCC mediated by VRC07-523LS for each of the viral strains upon pretreatment of NK cells with the combination of cytokines or no cytokine control (n=6-16). Statistical significance between no cytokine and IL12/IL15/IL18 was determined by two-way ANOVA and Wilcoxon signed rank test with hypothetical value set to 0 for each condition. Open symbols represent female and closed symbols male donors. ( B ) Spearman correlation between the percentage of ADCC and surface Env binding for the indicated antibodies and NK cells pretreated with no cytokine (top) or IL12/IL15/IL18 (bottom). Each virus is indicated with a different color (n=4-6).

Article Snippet: Ten cytokines were measured using Quanterix SP-X Corplex Cytokine Panel (IFN-γ, IL-1β, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, IL-22, TNF-α) (cat# 85-0329).

Techniques: Activation Assay, Binding Assay, Control, Virus

Journal: Scientific Reports

Article Title: Optimization Strategies for Bruch’s Membrane Opening Minimum Rim Area Calculation: Sequential versus Simultaneous Minimization

doi: 10.1038/s41598-017-14284-1

Figure Lengend Snippet: Principle of the BMO-MRA calculation in Spectralis SP-X VWM software. Figure 1a Graphical model; between every two B-scans, the local BMO-based minimum rim area is calculated as a trapezoid covering the minimum area between the internal limiting membrane (ILM) and Bruch ’ s membrane opening (BMO); Global and sectorial BMO-MRA values are determined by the addition of respective local minimum rim area results; model displays extreme tilt between two segments for purpose of illustration; BMO, Bruch ’ s membrane opening; BMO-MRA, BMO-based minimum rim area. (Note: graphical model modified from a version published previously ; (figure created with Microsoft Powerpoint of Microsoft Office Plus 2010, Microsoft Corporation).

Article Snippet: Spectralis SP-X VWM software is exclusive property of Heidelberg Engineering GmbH, Heidelberg, Germany.

Techniques: Software, Membrane, Modification

Journal: Scientific Reports

Article Title: Optimization Strategies for Bruch’s Membrane Opening Minimum Rim Area Calculation: Sequential versus Simultaneous Minimization

doi: 10.1038/s41598-017-14284-1

Figure Lengend Snippet: Principle the BMO-gMRA calculation in Spectralis SP-X VWM software. Graphical model; The gMRA surface is locally discretized in each SD-OCT star scan sector i as two adjacent triangles with areas ∆ i 1 and ∆ i 2. While BMO positions remain fixed during optimization, the position of BMO-ILM connections (i.e., the arrow in the OCT image) is optimized along the ILM segmentation line of the star. In this approach, global optimization is performed in all (typically 48) degrees of freedom simultaneously. BMO, Bruch’s membrane opening; BMO-gMRA, BMO-based globally optimized minimum rim area (Figure created with Microsoft Powerpoint of Microsoft Office Plus 2010, Microsoft Corporation).

Article Snippet: Spectralis SP-X VWM software is exclusive property of Heidelberg Engineering GmbH, Heidelberg, Germany.

Techniques: Software, Membrane