Fig. 3

HTLV-1∆CTCF virus immortalizes primary T-lymphocytes. 729 HTLV-1 producer cell clones were generated by nucleofection of 729.B cells with 2 ug of HTLV-1, HTLV-1∆CTCF, HTLV-1p12Stop proviral plasmid clones followed by stable cell selection via G418 treatment and subsequent limiting dilution single cell cloning. 729 HTLV-1 producer cell clones were then irradiated and functionally assessed via p19 Gag ELISA. a p19 Gag production was comparable between HTLV-1, HTLV-1∆CTCF, HTLV-1p12Stop producer cell clones. Irradiated producer cell clones (10⁶) were then cocultured in 24-well plates with freshly isolated hPBMCs (2 × 10⁶) to assess in vitro hPBMC immortalization capacity. b Viable cells were counted at weeks 0, 1, 2, 3, 4, 6, 8, 10, 14, and 16. HTLV-1, HTLV-1p12Stop, and HTLV-1∆CTCF all maintained hPBMC immortalization capacity. c Supernatant was collected and p19 Gag production was measured at weeks 3, 6, 10, and 14. HTLV-1, HTLV-1p12Stop, and HTLV-1∆CTCF displayed comparable p19 Gag production. For figures B and C, the mean (symbols) and standard deviation (error bars) was determined from three random, independent samples from each time point