Figure 3
From: Subtype-associated differences in HIV-1 reverse transcription affect the viral replication
Presence of the Gag and Pol domains from HIV-1 subtype C correlates with decreased level of virus replication. A - Kinetics of replication (solid lanes) and cytopathicity (dash lanes) of the backbone NL4-3 and chimeric NL-pol(1084) viruses in Sup-T1 cells. The cells (1 × 106) were incubated with virus suspensions (0.01 pg of p24CA per cell) and then cultured in a fresh culture media. Ninety percent of the volume of cell suspensions were harvested every 3 to 4 days, and replaced with uninfected cells. HIV-1 p24CA levels were detected in culture supernatants at the indicated days after infection. Cell viability was measured in cell suspensions using trypan blue staining. Each curve indicating p24CA concentration in the culture media represents the mean data of two independent experiments. Error bars show the standard error. Each curve indicating cell viability represents data of one experiment. B - Kinetics of replication (left panel) and cytopathic effect (right panel) of the indicated NL4-3-based viruses in Sup-T1 cells. Infection with virus clones and cultivation of infected cells were performed as described in A. The p24CA curves represent the mean data ± SE from two independent experiments. The curves indicating cell viability represent data from one experiment. C - Kinetics of replication and cytopathic effect of the backbone 1084i and chimeric 1084ipolL(NL) viruses in U87.CD4.CCR5 cells. Each viral inoculum (MOI = 0.05) was added to 0.25 × 106 cells. HIV-1 p24CA concentrations and cell viability were monitored at the indicated days. Each point represents mean p24CA level from two independent experiments. Error bars show the standard error. Each point indicating cell viability represents data of one experiment. D - Kinetics of replication (solid lanes) and the cytopathicity (dash lines) of the backbone NL4-3 and chimeric NL-polL(1084) viruses in Sup-T1 cells. Infection with virus clones and cultivation of infected cells were performed as in A. Each curve indicating p24CA concentration represents the mean data ± SE of two independent experiments. Each curve indicating cell viability represents data of one experiment. E - Syncytia formation by the Sup-T1 cells infected with the indicated virus strains. Live cells from the experiment described in A and B, maintained in 1 ml of culture medium, were subjected to phase-contrast microscopy on the indicated days after infection. One of ten representative images for each time point is shown.