Follicular Dendritic Cells Retain Infectious HIV in Cycling Endosomes
Fig 2
Human FDCs isolated from HIV positive subjects on ART treatment retain HIV in a Tf positive compartment.
(A) Cultured human FDCs from HIV+ subjects on ART were stained for Tf (green), HIV (p24, red), Lamp-1 (blue) and Hoechst (cyan). (inset) A single vesicle was enlarged and only the red (HIV) channel was shown. The yellow line was used for line profile analysis, which measures the fluorescent intensity over that line in all channels. Line profiles were also made in the z direction to ensure co-localization of HIV+ vesicle in the X, Y and Z direction with Tf or Lamp-1. (B) Quantification of line profile measurements of HIV positive vesicles. The vast majority of HIV containing vesicles were positive for Tf (~80%), while only ~5% was positive for Lamp-1 ****P<0.0001 (Two-way ANOVA) n = 2 (2 subjects, 4 replicates each). (C) FDCs and FRCs from HIV positive individuals on ART treatment and HIV negative subjects were cultured and viral RNA levels were quantified. RNA levels are represented per well. FRC and FDC were isolated from the same subjects to control for contaminating cells. Each data point presents an individual subject. ****P<0.0001 (Two-way ANOVA, multiple comparisons) n = 4 (4 subjects, 3 replicates).