Galpin/DA/N2 IIIBenv 
Mus dunni cocultivations are performed to amplify any low level RCR in a test 
cell line. Producer cell lines are cocultivated with Mus dunni cells for 15 days, and the 
resulting supernatants are tested using MdC, MdH, or extended S+L* assays. 
The producer cell line DA/N2 IIIBenv. 2C3 was tested by Mus dunni cocultivation 
for the generation of RCR (Figure 7). Positive controls are the DA/N2 IIIBenv. 2C3 
cells cocultivated with Mus dunni cells and spiked with increasing numbers of Md/MA 
cells ( Mus dunni cells infected with the hybrid MoMLV-4070A amphotropic virus 
MA). These data show no detection of RCR in the producer cells. The controls 
indicate that approximately one cell infected with RCR can be detected in this test 
system. We have performed similar testing on the producer cell line before banking, 
on the banked cell line, and on cells post production. These have given uniformly 
negative results. 
3.0 PRECLINICAL STUDIES 
Preclinical studies have utilized three animal models to test the immunobiological 
activity of HIV-IT (V). The study models were normal mice. Rhesus monkeys, and baboons. 
In addition, in vitro immune stimulation studies have been conducted with lymphocytes 
obtained from HIV- 1 -infected individuals. Animal studies have examined and demonstrated 
the ability of HIV-IT (V) to stimulate HIV-1 envelope-specific CTL and antibody responses 
in vivo in the animal models. No toxicological effects have been observed in animals 
receiving HIV-IT (V) treatment. 
These preclinical studies involving direct administration of HIV-IT (V) provided the 
foundation for the planning of clinical trials in humans. The study parameters were: dose, 
number of injections, types of induced immune responses, and the establishment of human 
CTL monitoring systems. 
Recombinant DNA Research, Volume 17 
