representative experiment is shown in Fig. 1. It is important to 
note that a non-specific inhibitory effect is seen when cells are 
grown in the presence of G-418. This effect has been described 
by many laboratories. In addition to this effect, Rev M10 shows 
the ability to inhibit productive viral replication compared to 
cells containing the frameshift Rev M10 negative control. 
PROTECTIVE EFFECTS Of REV M 10 IN PRIMARY HUMAN T CELLS 
Rev Ml 0 neo 
fsRevMlO neo 
I I P8L 
Figure 1. Freshly isolated PBMC's were stimulated for 48 hours at 
37 °C with 5/ig/ml PHA. On day 3 and 5, cells were infected for 8 
hrs with ipe rip supernatants, (plus 5/xg/ml polybrene) containing 
the PLJ-Rev M10 neo or frameshift PLJ-fsRev MlO(neo) retrovirus. 
Cells were subsequently selected with 300//g/ml G418 for a 
further 8 days. On day 8 , 5 x 10 6 cells in 0.5ml of medium were 
incubated with approximately 5 x 10 4 TCID 50 of HIV 
bru 
or a 
freshly passaged clinical isolate for 2hrs at 37 c . Cells were 
washed and resuspended at 1 x 10 6 /ml. Day 7 post HIV infection, 
duplicate culture supernatants ( 10 ^ 1 ) were assayed for reverse 
* 
transcriptase activity. 
Recombinant DNA Research, Volume 18 
[33] 
