Figure 5
Human adult primary microglia recapitulate engagement with hBORGs, supports HIV-1 infection and produces inflammatory cytokines. (A) Schematic diagram of the experimental design is depicted. Primary adult brain microglia (0.5 × 106 cells) were infected with HIV-1 (panel a, green) or mock-infected, membrane-labeled (red) and were added to hBORGs for overnight. Scale is 100 μm. After 24 h, microglia incorporated-hBORGs were transferred to a new plate and maintained for an additional 30 days for further analyses. Image (panel b) depicts the infiltration of primary adult brain microglia into hBORGs. Infected and uninfected microglia were membrane-labeled (red), incorporated into hBORGs and imaged by confocal microscopy. HIV-infected microglia (green, indicated with black arrow) and uninfected microglia (red) was incorporated in hBORGs on day 1 post coculture (panel b). Scale bar is 100 μm. (B) RT-qPCR assessment of HIV-1 gag mRNA copies in primary MG-hBORGs at day 30 post infection (N = 3). (C) Representative cumulative HIV-1 virus titer in supernatants from HIV-infected MG-hBORGs where MG-hBORGs were developed using NPCs from two different donors. Kinetics of cumulative levels of (D) TNF-α and (E) IL-1β released from HIV-1 infected, mock-infected MG-hBORGs and BORGs without microglia during the course of infection measured by ELISA (N = 4). **p < 0.01, * p < 0.05.
