independently on human melanoma and renal cell carcinoma in 
vitro, and confirmed by direct injection into melanoma or other 
tumors in vivo prior to use. Each component, the liposome 
preparation and the DNA, will be tested for contaminants and 
toxicity and used according to previously established guidelines 
from the FDA. The liposome solution and DNA will be aliquoted in 
individual sterile vials mixed under sterile conditions as 
described above. 
For direct injections of the HLA-B7 plasmids, escalating 
doses will be studied in this phase I study. Four groups (3 
patients each) will be studied sequentially with at least 1 month 
of observation prior to evaluation of the next group. Patients 
in each group will receive intratumor injections. Group I will 
receive 3 injections of 0.2 ml within the same nodule (3 j ig of 
DNA + 4.5 riM DMRIE/DOPE) . Group II will receive the same 
treatment with a 10-fold higher concentration of DNA liposome 
complex. Group III will receive 100-fold higher dose, and Group 
IV will receive 1000 x higher amount. We are currently 
evaluating whether pre-treatment with low dose cytoxan can 
improve the anti-tumor response by eliminating suppressive T 
cells. If this approach is helpful, it will be included as part 
of the protocol. 
For catheter-based gene delivery, the same dose escalation 
will be used, except a single 0.6 ml injection into the end 
artery which perfused an isolated nodule will be used with an 
occlusion balloon catheter. In murine and porcine models, the 
highest treatment exceeded these proposed doses by 100-fold and 
Recombinant DNA Research, Volume 17 
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