CXCL8 increases HIV-1<i>p24</i> and 2-LTR circles in HIV-1-infected primary human microglia.

<p>Primary human microglia were infected with HIV-1_ADA or HIV-1_JRFL (MOI 0.1) overnight, followed by treatment with 50 ng/ml CXCL8. (<b>A</b>) Supernatants were collected 2 weeks post-infection and HIV-1<i>p24</i> levels were measured by ELISA. (<b>B</b>) DNA was isolated and 2-LTR circle junctions were amplified by real-time PCR. (<b>C–E</b>) MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) reagent was added to microglia and incubated for 15 minutes at 37°C. Results are expressed as mean ± SEM, analyzed by one-way ANOVA and Newman-Keuls post-test for multiple comparisons. Phase contrast pictures showing (<b>C</b>) cell morphology of uninfected primary human microglia. (<b>D</b>) Multinucleated giant cells formed with HIV-1_ADA infection (<b>E</b>) HIV-1_ADA infection and CXCL8 treatment. (<b>F-H</b>) Expression of CD68 (green) and HIV-1<i>p24</i> (red) was measured by immunocytochemistry 2 weeks post-infection with HIV-1_ADA. Nuclei were stained in blue by DAPI. (<b>F</b>) Uninfected primary human microglia (<b>G</b>) co-localization (yellow) of CD68 and HIV-1<i>p24</i> in HIV-1_ADA-infected microglia. (<b>H</b>) HIV-1<i>p24</i> expression and multinucleated giant cells in HIV-1_ADA-infected microglia. Arrows represent multinucleated giant cells. Original magnification 200X.