Figure 1
Astrocyte HIV-1 YU-2 -transfection increases CD38 expression. Cultured human astrocytes were transfected with HIV-1YU-2 plasmid and mock-transfected controls were maintained in parallel. RNA was isolated 24 h post-transfection and assayed for CD38 levels. (A) By real-time-PCR assay, significantly higher CD38 mRNA levels were detected in HIV-1YU-2-transfected astrocytes as compared to mock controls (p < 0.05). (B) HIV-1YU-2-transfected astrocytes showed significantly increased CD38 protein levels as compared to mock controls when assayed by western blot and densitometry analyses of whole cell protein lysates (p < 0.01), 5 days post-transfection. IL-1β (20 ng/ml)-treated astrocytes were maintained as positive controls for increase in CD38 mRNA and protein levels. Graph shows representative data from two independent donors. Expression of CD38 and HIV-1p24 was measured by immunocytochemistry 5 days post-transfection (C-F). Nuclei were stained in blue by DAPI in all panels. (C) Mock, GFAP-positive astrocytes (red) with no HIV-1p24 expression. (D) Co-localization (yellow) of GFAP (green) and HIV-1p24 (red) in HIV-1YU-2-transfected astrocytes. Transfection efficiency was routinely 90% as assessed by GFAP, HIV-1p24 co-localization (data not shown) (E) Basal CD38 expression (green) and no HIV-1p24 expression in mock control. (F) Increased CD38 expression (green) and co-localization (yellow) with HIV-1p24 (red) in HIV-1YU-2-transfected astrocytes. Original magnification x200.