Figure 2.
In vivo neutralization and in vitro antiviral properties of 7-269 IgA bNAb. (A) In vivo neutralization activity of human 7-269 bNAb in HIV-1–infected hu-mice. Graphs compare HIV-1 plasma viremia in ART-treated BRGS hu-mice receiving a single i.p. injection (0.5 mg) of 7-269 (n = 5) or mGO53 control (n = 8) IgA antibodies 24 h before ART interruption. (B) Kaplan–Meier analysis of the in vivo effect of 7-269 IgA on viral rebound (VR) following ART interruption. Groups were compared using log-rank (Mantel–Cox) test. (C) Dot plots comparing the percentage of transcytosis (top) and post-transcytosis infectivity (as RLU, bottom) of AD8 and CH058 virions alone (No Ab), in the presence of non–HIV-1 mGO53 control (Ctr) and 7-157 and 7-269 IgA antibodies. Mean values of triplicate values from two independent experiments are shown. Antibody groups were compared to the No Ab group using Mann–Whitney test. **, P < 0.01; ns, not significant. (D) Heatmap comparing the percentage of target cells infected by lab-adapted (AD8, YU2) and T/F (CH058, CH077, THRO) viruses, and bound by selected IgG antibodies (IgG+Gag+) as measured by flow cytometry. Mean values from two independent experiments are shown. (E) Binding of IgA bNAbs to HIV-1–infected cells. Flow cytometric histogram (top left) shows the reactivity of 7-269 IgA antibodies to Gag+ infected target cells. Heatmap (bottom left) shows the same as in D but for IgA antibodies. Graphs (right) show antibody binding titrations to AD8- and CH058-infected cells, measured as percentage IgA+ among Gag+ cells by flow cytometry. Mean values from two independent experiments are shown. FI, fluorescence intensity. (F) ADCC potential of pt7 bNAbs expressed as IgG antibodies against AD8-infected target cells. Flow cytometric histogram (left) comparing the percentage of FarRed+Gag+ cells among infected CEM.NKR cells incubated with 7-269 IgG or non–HIV-1 isotype control (mGO53, IgGCtr) in the presence of human NK cells or not. Histogram (right) comparing the percentage ADCC of AD8-infected targets incubated with selected IgG and IgA antibodies. PGT128 and 10-1074 are positive controls, and mGO53 is the negative control. Dots correspond to means of percentage ADCC values measured in duplicate for each NK-isolated human donor (n = 4 and n = 5 for IgAs and IgGs, respectively). (G) Competition ADCC. Heatmap (left) showing competition for ELISA binding to BG505 SOSIP.664 of selected HIV-1 bNAbs. Lighter colors indicate stronger inhibition; dark blue indicates no competition. Dot plot (right) comparing the percentage ADCC of AD8-infected targets by 7-319 and PGT128 IgG antibodies in the presence of 7-269 IgA used as competitor. Dots correspond to means of percentage ADCC values measured in duplicate for each NK-isolated human donor (n = 8). Groups were compared using two-tailed Wilcoxon test. **, P < 0.01. (H) Bar graphs showing the ADCP activity of selected IgA (blue) and IgG (red) antibodies expressed as normalized PS (nPS). Each dot corresponds to a healthy donor of primary monocytes (n = 4 or 8) and presents the mean of duplicate nPS values. d10-1074, dimeric 10-1074 IgA; 10-1074G, 10-1074GASDIE mutant antibody. Antibody groups were compared to the control (Ctr) group using two-tailed Mann–Whitney test. Only P values <0.05 are indicated: *, P < 0.05; **, P < 0.01. (I) Monocyte- and neutrophil-mediated ADCC potential of 7-269 IgA bNAb against AD8-infected target cells. Bar graphs comparing the percentage of FarRed+Gag+ cells among infected CEM.NKR cells incubated with 7-269 IgA1, 10-1074 IgA1, or non–HIV-1 isotype control (mGO53 IgA1, Ctr) in the presence of human monocytes or neutrophils with target:effector ratios of 1:5, 1:10, and 1:20. Each dot corresponds to a healthy donor of primary monocytes and neutrophils (up to n = 4) and presents the mean of duplicate values. P values comparing HIV-1 antibodies with the control (Ctr) using two-tailed Mann–Whitney test were not significant.

In vivo neutralization and in vitro antiviral properties of 7-269 IgA bNAb. (A) In vivo neutralization activity of human 7-269 bNAb in HIV-1–infected hu-mice. Graphs compare HIV-1 plasma viremia in ART-treated BRGS hu-mice receiving a single i.p. injection (0.5 mg) of 7-269 (n = 5) or mGO53 control (n = 8) IgA antibodies 24 h before ART interruption. (B) Kaplan–Meier analysis of the in vivo effect of 7-269 IgA on viral rebound (VR) following ART interruption. Groups were compared using log-rank (Mantel–Cox) test. (C) Dot plots comparing the percentage of transcytosis (top) and post-transcytosis infectivity (as RLU, bottom) of AD8 and CH058 virions alone (No Ab), in the presence of non–HIV-1 mGO53 control (Ctr) and 7-157 and 7-269 IgA antibodies. Mean values of triplicate values from two independent experiments are shown. Antibody groups were compared to the No Ab group using Mann–Whitney test. **, P < 0.01; ns, not significant. (D) Heatmap comparing the percentage of target cells infected by lab-adapted (AD8, YU2) and T/F (CH058, CH077, THRO) viruses, and bound by selected IgG antibodies (IgG+Gag+) as measured by flow cytometry. Mean values from two independent experiments are shown. (E) Binding of IgA bNAbs to HIV-1–infected cells. Flow cytometric histogram (top left) shows the reactivity of 7-269 IgA antibodies to Gag+ infected target cells. Heatmap (bottom left) shows the same as in D but for IgA antibodies. Graphs (right) show antibody binding titrations to AD8- and CH058-infected cells, measured as percentage IgA+ among Gag+ cells by flow cytometry. Mean values from two independent experiments are shown. FI, fluorescence intensity. (F) ADCC potential of pt7 bNAbs expressed as IgG antibodies against AD8-infected target cells. Flow cytometric histogram (left) comparing the percentage of FarRed+Gag+ cells among infected CEM.NKR cells incubated with 7-269 IgG or non–HIV-1 isotype control (mGO53, IgGCtr) in the presence of human NK cells or not. Histogram (right) comparing the percentage ADCC of AD8-infected targets incubated with selected IgG and IgA antibodies. PGT128 and 10-1074 are positive controls, and mGO53 is the negative control. Dots correspond to means of percentage ADCC values measured in duplicate for each NK-isolated human donor (n = 4 and n = 5 for IgAs and IgGs, respectively). (G) Competition ADCC. Heatmap (left) showing competition for ELISA binding to BG505 SOSIP.664 of selected HIV-1 bNAbs. Lighter colors indicate stronger inhibition; dark blue indicates no competition. Dot plot (right) comparing the percentage ADCC of AD8-infected targets by 7-319 and PGT128 IgG antibodies in the presence of 7-269 IgA used as competitor. Dots correspond to means of percentage ADCC values measured in duplicate for each NK-isolated human donor (n = 8). Groups were compared using two-tailed Wilcoxon test. **, P < 0.01. (H) Bar graphs showing the ADCP activity of selected IgA (blue) and IgG (red) antibodies expressed as normalized PS (nPS). Each dot corresponds to a healthy donor of primary monocytes (n = 4 or 8) and presents the mean of duplicate nPS values. d10-1074, dimeric 10-1074 IgA; 10-1074G, 10-1074GASDIE mutant antibody. Antibody groups were compared to the control (Ctr) group using two-tailed Mann–Whitney test. Only P values <0.05 are indicated: *, P < 0.05; **, P < 0.01. (I) Monocyte- and neutrophil-mediated ADCC potential of 7-269 IgA bNAb against AD8-infected target cells. Bar graphs comparing the percentage of FarRed+Gag+ cells among infected CEM.NKR cells incubated with 7-269 IgA1, 10-1074 IgA1, or non–HIV-1 isotype control (mGO53 IgA1, Ctr) in the presence of human monocytes or neutrophils with target:effector ratios of 1:5, 1:10, and 1:20. Each dot corresponds to a healthy donor of primary monocytes and neutrophils (up to n = 4) and presents the mean of duplicate values. P values comparing HIV-1 antibodies with the control (Ctr) using two-tailed Mann–Whitney test were not significant.

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