Figure 3. ACV suppresses HIV-1 in HHV-free MT-4 cell cultures either in the presence of HHV-6 infected cells or as monophosphorylated prodrug.
a. HIV-1LAI.04 -infected MT4 cells were co-cultured with HHV-6B-infected MT4 cells in a ratio 10:1. Presented are the distributions of HHV-6B (upper panel) and HIV-1 LAI.04-infected cells (lower panel) as measured by flow cytometry at day 3 post-HIV-1 infection, in cultures untreated or treated with various concentrations of ACV. Note that the fraction of both HHV-6B- and HIV-1-infected cells is reduced in a dose dependent manner by ACV treatment.
b. HIV-1LAI.04 -infected MT4 cells were co-cultured with HHV-6B infected MT4 cells. HIV-1 replication was monitored as in Fig.1a. Note that the replication of HIV-1 in HHV-6B/HIV-1-infected co-cultures is suppressed by ACV treatment in a dose dependent manner.
c. The compound acyclovir-[1-naphthyl (methoxy-L-alaninyl)]phosphoramidate (Cf2649) has been synthesized as described in Supplemental data.
d. HIV-1 LAI.04 -infected HHV-free MT4 were treated with various concentrations of the ACV monophosphorylated prodrug Cf2649. Presented are the kinetics of HIV-1LAI.04 replication, monitored by measuring p24gag accumulated in culture media at day 2, 3, 4 and 6 post-infection. Presented data are representative of two experiments. Note that compound Cf2649 suppresses the replication of HIV-1LAI.04 in a dose dependent manner.
e. HIV-1LAI.04 -infected MT4 cells not infected with any HHVs were treated with various concentrations of the compound Cf2649. The cumulative release of p24gag into culture media over 6 days of culture treated with various concentrations of the compound Cf2649 is presented as fraction of the p24gag production in the untreated cultures. Presented data are representative of two independent experiments. Note that compound Cf2649 suppresses the replication of HIV-1LAI.04 in a dose-dependent manner.