Figure 1

M48U1 has a memory effect on cell-associated virus. A) The Trojan Horse transmission concept was modeled in vitro using HIV-infected PBMCs which were treated for 24 h with antiretrovirals (ARVs) from different classes at 100x EC50 concentrations. Subsequently, virus production was quantified using a Gag p24 ELISA. To mimic escape from vaginal drug pressure, the infected cells were then incubated in ARV-free medium for three consecutive periods of 24 h. Infectivity of newly produced virions was determined after each period by titration in TZM-bl cells and PBMCs using equal amounts of p24. B) Production of Gag p24 by infected PBMCs during 24 h treatment with eleven ARVs: the non-nucleoside reverse transcriptase inhibitors (NNRTIs; blue) UC781 and TMC120, the nucleotide reverse transcriptase inhibitor tenofovir (PMPA; red), the entry inhibitors (grey) maraviroc (MVC), T-20, C34-chol, griffithsin (GRFT) and M48U1, the protease inhibitors (green) lopinavir (LPV) and saquinavir (SQV), and the integrase inhibitor raltegravir (RAL). Values are expressed relative to untreated control cultures and represent the mean +/− SEM of at least three independent experiments, each carried out in triplicate. C) Bal infectivity of de novo produced virus by pretreated infected PBMCs in absence of drug during three consecutive periods of 24 h. Values are expressed relative to untreated control cultures and represent the mean +/− SEM of at least three independent experiments, each carried out in triplicate. D) Infectivity of de novo produced transmitted/founder viruses REJO and THRO (subtype B) and Bal by M48U1 treated infected PBMCs in absence of drug during three consecutive periods of 24 h. Values are expressed relative to untreated control cultures. E) Infectivity of wild type (black) and M48U1-resistant (red) Bal virus produced during a 24 h period in absence of drug by pretreated infected PBMCs. Values represent the 50% Tissue Culture Infective Dose (TCID50)/ml as measured in one experiment carried out in triplicate.