Tipranavir

Washed human platelets were diluted to 1.1×10⁸ platelets/mL in TSS and 300 μL aliquots were plated into each well of a 96-well plate. Platelets were treated with ARVs or vehicle (PBS or <0.1% DMSO) for 30 minutes at 37°C as follows: ritonavir 5 μM, abacavir 20 μM, darunavir 10 μM, raltegravir 20 μM, lamivudine 20 μM, tipranavir 25 μM, and atazanavir 5 μM. At 30 minutes, the 96-well plates were centrifuged at 1,200 x g for 10 minutes. Platelet supernatants were collected into a separate 96-well plate for storage and stored at −20°C until used to detect TXB₂ and PGE₂ using enzyme immunoassays (Cayman Chemical). The following ARV reagents were obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: ritonavir (Cat. # 4622), darunavir ethanolate (Cat. # 11447), abacavir (Cat. # 4680), lamivudine (Cat. # 8146), and raltegravir (Cat. # 11680); tipranavir (Cayman Cat. #21028) and atazanavir (Cayman Cat. # 11733) were purchased from Cayman.