Fig. 5.

Replication properties of the B1 and B2 mutant viruses. Relative virus production (A) and infectivity (B) of the mutant viruses. Viruses were harvested 36 hours post-transfection and quantified by p24 CA ELISA; equal amounts of viruses were used to infect TZM-bl cell and luciferase activities were measured. Measurements from wild-type NL4-3 (WT) were defined as 100%. Results from three independent experiments are summarized; error bars indicate standard deviations. Replication of the wild-type and the B1 (C) and B2 (D) mutant viruses. Representative replication kinetics of the mutant virus are shown on the left whereas those from NL4-3 virus performed in parallel are shown on the right. Equal amounts of viruses were used to infect CEM cells, supernatants were harvested at 2, 4, 6, 8, 10 or 12 days post-infection, and virus production was measured by p24 ELISA. Three independent experiments were performed for each mutant; one representative replication curve for each mutant virus is shown. Replication fitness of the B1 (E) and B2 (F) mutant virus measured by competition assay. Wild-type and mutant viruses were mixed at the ratios of 1:9 (top panels) and 1:1 (bottom panels). Results from two independent experiments using CEM cells are summarized in the left panels with error bars indicating standard deviation, whereas results from one representative experiment using PBMCs are shown in the right panels. PBMCs from three donors were used in three independent experiments. Day zero samples are the results from the input virus of each infection. X-axis: Days post infections. Y-axis: Proportions of wild-type and mutant virion RNA released in the supernatant.