Figure 5

Detection of systemic infection with rubella vectors by RT-PCR. Group 1 macaques were immunized with live rubella vectors given IM in the quadriceps, and gingival mouth swabs were collected at 0, 1, and 2 weeks after immunization. Viral RNA containing the MPER insert was amplified by RT-PCR, and the products were resolved in 2% agarose gel. Lane 7 shows the PCR band at 347 bp for a plasmid with the same MPER insert as in the viral vector (upper panel). No virus was detected in pre-injection samples (lanes 1–3, representing J6L, V200, and V584). One week post infection, two of three animals were positive for virus (lanes 4 and 6). By week 2, both of these were resolved (lanes 8, 10), and the third animal was positive (lane 9). Similar results are shown for RT-PCR of the BC-sGag2 insert (lower panel), with a positive band at 494 bp for the plasmid control (lane 7). All animals were negative at the start (lanes 1–3), two of three were positive at week 1 (lanes 5 and 6), and both had cleared by week 2 (lanes 9 and 10). We detected no signal for BC-sGag2 from one of the macaques (J6L) at either time point.