FIG 6.
HIV-1 tat and gp120 disrupt tonsil epithelial junctions and facilitate HCMV infection. (A) Tonsil tissue explants were treated with active or inactive HIV-1 tat and gp120 proteins for 5 days; culture medium was changed daily to add fresh proteins. After each day, one set of explants was immunostained for occludin (red). (B) One set of tonsil explants treated with active tat and gp120 for 3 days was infected with 3 × 105 IU of HCMV VR1814; after 2 days, tissues were coimmunostained for HCMV gB (green) and occludin (red). Cell nuclei were stained with DAPI (blue), and cells were analyzed by fluorescence microscopy. Magnification, ×400. EP, epithelium; LP, lamina propria. (C) Tonsil explants from two independent donors were treated with active tat and gp120. After 3 days, explants were infected with 3 × 105 IU of HCMV VR1814. After 2 more days, tissues were immunostained for HCMV gB; gB-expressing cells were counted, and values are presented as a percentage of infection. (D) Tonsil tissues from six donors (#1 to #6) were treated with HIV-1 active or inactive tat and gp120 for 3 days. Untreated tissues served as a control. Tissues were then infected with HCMV VR1814 for 3 days. Tissues were immunostained for HCMV gB, and HCMV gB-expressing cells were quantitatively evaluated. (A and B) Data are means and SD of triplicate values. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 (compared with the controls treated with inactive tat+gp120).