Figure 5
Selective induction of HIV-1 replication (or expression of VSV-G pseudotyped virus) in M12 MDM stimulated by allogeneic PHA blasts or their supernatant and its selective prevention by lamivudine/3TC. (A) Kinetics of RT activity. PHA blasts upregulated virus replication in M12 MDM, but not in CTR or M1-MDM; the arrowheads indicate the timing of PHA blast addition to the infected MDM cultures. Mean ± SEM of 7 independent donors is reported. Comparison of different time points between M1 vs. M12 plus PHA blasts was performed using One-way ANOVA. ***p < 0.001. The addition of lamivudine/3TC (1 µM - black arrowheads) did not substantially affect virus production in CTR and M1-MDM, but fully prevented HIV-1 replication in M12 MDM. (B) Left panel: kinetics of RT activity; right panel: RT activity ratios relative to unstimulated M12 MDM calculated at peak virus replication in the different conditions (n = 2). The arrowhead indicates the timing of stimuli addition to the infected MDM cultures. (C) CTR, M1-MDM and M12 MDM were infected with a VSV-G pseudotyped virus expressing eGFP. Thirteen days after infection the cells were incubated with Blast Sup for 72 h before determining the levels of eGFP expression. Both M1-MDM and M12 MDM showed reduced levels of GFP expression vs. CTR cells, but only M12 MDM were significantly enhanced in terms of virus expression by incubation with Blast Sup (p < 0,05 by T-test, n = 3; see also Fig. 9S).
