4.0 PROTOCOL SCHEMA 
Tumor and SXln Harvest 
Tumor 
Prepare and Freeze Single 
Cell Tumor Suspension 
Thaw 
Separate viable cells 
S*in 
Establish fibroblast line 
4 
Transduce 
Insert IL-4 gene 
4 
Expand 
Select with G418 
i 
IL-4 Elisa and bioassav 
Activity in culture supernatant, 
when cell population reaches 5 X 10 7 
4 
Harvest 
2 X 10 7 Fibroblasts 
Prepare Vaccine 
5 X 10 6 Tumor Cells 
+ 
<10 7 Fibroblasts; < 10 5 U IL-4/D 
+ 
5000 rads 
4 
Primary vaccination-dav 1 
5 sites: 10 7 nontransduced , control fibroblasts + 5 X 10 6 tumor cells 
graded doses of fibroblasts transduced with the IL-4 gene + 5 X 10 6 tumor 
cel^s 
Biopsv-dav IS 
All sites 
Secondary vaccination-dav 15 
5 sites: all with highest tolerated dose of 
IL-4- transduced fibroblasts + 5 X 10 6 tumor cells 
i 
Biopsy 
Days 16, 17, 19, 22, and 29 
4 
Clinical and radiographic assessment-dav 60 
Recombinant DNA Research, Volume 16 
[505] 
