Figure 8.

HIV-1 TAR inhibitors lower proinflammatory cytokine production in astrocytes. U937 and U1 cells were cultured for 5 days. U1 monocytes were differentiated into MDMs using PMA (100 nM) and treated ± cART cocktail (10 µM of darunavir, emtricitabine, tenofovir, and 5 µM of ritonavir) for 5 days. EVs were isolated using ultracentrifugation and assessed for concentration and particle size using ZetaView NTA to equilibrate EV concentrations. EVs were used to treat astrocytes (CCF-STTG1; 1 × 10⁶) at a 1:10,000 concentration (cell/EVs ratio). CCF-STTG1 cells were treated with ± EVs from U937 cells, as well as U1 MDMs that were treated with cART (10 µM of darunavir, emtricitabine, tenofovir, and 5 µM of ritonavir) at a 1:10,000 concentration (cell/EVs ratio), and ±titration of TAR inhibitors (100, 10, and 1 nM; 103FA, 111FA, and Ral HCL) for 3 days. EVs from supernatants were enriched using NT80/82 particles and analyzed for the presence of IL-6 and Actin using Western blot.