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. 2019 Apr 9;27(2):572–585.e7. doi: 10.1016/j.celrep.2019.03.032

Figure 1.

Figure 1

Capture of HIV-1 Env-Reactive Mucosal B Cells

(A) Representative ELISA graph showing the reactivity of purified serum IgG (sIgG) from HIV-1-infected individuals (n = 5) against trimeric gp140, gp120, and gp41 proteins. Error bars indicate the SEM of duplicate values. Ctr+, positive control sIgG; pt3, patient 3 (Scheid et al., 2009); Ctr−, negative control sIgG; hd2, healthy donor 2 (Prigent et al., 2016).

(B) Neutralization activity of sIgG from HIV-1-infected donors (n = 5) measured by TZM-bl assay.

(C) Dot plots comparing the percentage of IgA+CD19+ and IgG+CD19+ cells in the IEL, LPL, and peripheral blood mononuclear cell (PBMC) compartments determined by flow cytometry as shown in Figure S1. Median values are indicated below.

(D) Dot plots comparing the distribution of B cell subsets in the IEL, LPL, and PBMC compartments determined by flow cytometry as shown in Figure S1. Percentage of mature naive (MN), resting memory (RM), activated memory (AM), and tissue-like memory (TLM) B cells and antibody-secreting cells (ASC) are shown.

(E) HIV-1 gp140-reactive B cell fluorescence-activated cell sorting (FACS). (Top) Cytograms show the gating strategy used for single-cell sorts of gp140-binding mucosal B cells. An example is given for patient COA21. (Bottom) Dot plots comparing the percentage of gp140-reactive IgA+ and IgG+ CD19+ lymphocytes from the IEL, LPL, and PBMC compartments determined by flow cytometry as shown in Figure S1.

(F) Pie charts show the distribution of single gp140-binding B cells captured by FACS with YU-2 gp140 trimers among IEL and LPL IgA+ and IgG+ B cell populations. The number of B cells captured is indicated in the center of each pie chart.

(G) Representative ELISA graph showing the reactivity of gp140-captured mucosal monoclonal antibodies (mAbs; n = 76) against YU-2 gp140 trimers. Error bars indicate the SEM of duplicate values. Dotted black and green lines represent the positive control 10-1074 and negative control mGO53 IgG antibodies, respectively. The proportion of high affinity anti-gp140 antibodies in each patients’ group is shown below in the pie charts. The total number of tested antibodies is indicated in the center of each pie chart.

See also Figures S1 and S2 and Tables S1 and S2.