Fig. 4: Detection of infused bNAbs in LN sections. | Nature Communications

Fig. 4: Detection of infused bNAbs in LN sections.

From: Administration of anti-HIV-1 broadly neutralizing monoclonal antibodies with increased affinity to Fcγ receptors during acute SHIVAD8-EO infection

Fig. 4

A Representative confocal microscopy staining to detect infused bNAbs in LN sections from monkeys that were treated with bNAbs early after SHIVAD8-EO challenge. bNAb staining was performed in one LN section per timepoint per animal. Inset (scale bar: 500 µm) shows CD20 (yellow) and cellular nucleic acids (JOPRO, blue) in the whole LN section; big image (scale bar: 50 µm) shows human Ig kappa light chain+ cells (pink), human Ig lambda light chain+ cells (turquoise), and cellular nucleic acids (blue); and small images (scale bars: 5 µm) show cellular nucleic acids (blue) with only human Ig kappa light chain+ cells (pink) (top left), only human Ig lambda light chain+ cells (turquoise) (top right), or an overlay of both (bottom). The big image and small image on the bottom right show pink and turquoise Surface objects assigned by Imaris for easier visualization of human Ig kappa light chain+ and lambda light chain+ cells. All other images show real staining signals. Quantification of total WT bNAb+ and total DEL bNAb+ cells (B) or VRC07-523-LS+, PGT121+, VRC07-523-LS/DEL+, and PGT121/DEL+ cells (C) in whole LN sections at weeks 2 (circles) and 8 (squares) post-challenge. N = 6 per group per timepoint. Each p = 0.0087, two-sided p-values. D Percentage of follicular VRC07-523-LS+, PGT121+, VRC07-523-LS/DEL+, and PGT121/DEL+ cells in LN sections at weeks 2 (circles) and 8 (squares) post-challenge. N = 6 per group per timepoint. P = 0.0260, two-sided p-value. Bar graphs show the mean and individual datapoints (BD). The Mann-Whitney test was used to detect significant differences in bNAb+ cell levels at each timepoint or between timepoints for each type of bNAb (WT or DEL) (B). The Mann–Whitney test was also used to detect significant differences between the levels of VRC07-523-LS+ and PGT121+ cells, VRC07-523-LS/DEL+ and PGT121/DEL+ cells, VRC07-523-LS+ and VRC07-523-LS/DEL+ cells, or PGT121+ and PGT121/DEL+ cells at each timepoint, or between timepoints for cells bound to each bNAb (C, D). N indicates the number of biological replicates (monkeys). Source data are provided in the Source Data file. kappa, human Ig kappa light chain; lambda, human Ig lambda light chain.

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