Figure 6.
MV LV B cell transduction of activated B cells results in eCD4-Ig production in activated B cells and plasmablasts and increasing production of eCD4-Ig in plasmablasts
(A) Schematic of the workflow of B cell studies performed. Supernatants were collected and assessed for eCD4-Ig-KiHR production on days 7 and 10 and for HIV-1 neutralization capacity on day 10. See materials and methods for culture condition information. B cells from donors 2 and 3. (B) eCD4-Ig-KiHR production from the listed (see legend) stably LV transduced B cells were normalized to the total number of live cells per day for each collection time. Data presented in dot plots are mean ± SD of triplicates. ns, not significant; ∗∗p < 0.01, ∗∗∗p < 0.01, ∗∗∗∗p < 0.0001. (C) eCD4-Ig-KiHR-containing supernatants from MV, but not VSV-G, pseudotyped LV transduced and differentiated plasmablasts can efficiently neutralize HIV-1 isolates. Supernatants were evaluated for eCD4-Ig-KiHR neutralization capacity from B cells. The neutralization assay, and HIV-1s used for evaluation, are described in Figure 4C. Paired one-sided Welch’s t test on IC50 HIV-1 neutralization assessment of n = 6 eCD4-Ig-WT and n = 6 eCD4-Ig-KiHR repeats, three repeats for each LV promoter, p < 0.022. Blue horizontal line marks IC50, and graph points show range and average of duplicates. For some samples, the lack of range is due to lack of variance between individual assays. IC50s were calculated from six dilution points of each supernatant of interest.