are submitted with this protocol. 
Following extensive review by investigational review committees of the. 
National Cancer Institute, National Heart, Lung and Blood Institute, the 
Recombinant DNA Advisory Committee (RAC) and the Food and Drug Administration 
we received permission to transfer TIL modified by insertion of the gene coding 
for neomycin resistance into cancer patients. The first patient was treated on 
May 22, 1989, and we have subsequently treated a total of seven patients with 
these gene modified TIL. Extensive studies have been conducted on the first 
five patients and they are presented in Tables 6 and 7 and Figure 1 (25). No 
toxicities of any kind could be attributed to the gene modification of the TIL. 
The expected toxicities associated with the concomitant administration of IL-2 
were seen. Patients received up to 1.45 x 10^^ gene transduced TIL popula- 
tions. The percent of cells transduced in these populations varied between one 
and 11%. In all cases, integration and expression of the NeoR gene was 
demonstrated. As shown in Figure 1, gene modified TIL could be detected at 
tumor deposits as long as 64 days after gene transduction. 
All safety studies performed in these patients showed no evidence of ex- 
posure to replication competent virus. 3T3 amplification and S+/L- assays 
revealed no replication competent virus present in the TIL at the time they 
were infused. Polymerase chain reaction analysis for the viral envelope gene 
and reverse transcriptase assay of the gene-modified TIL culture were also 
negative. Western blot analyses of patient serum at varyious times after cell 
administration were all negative for evidence of exposure to virus as well. 
These studies demonstrated that gene modification of the TIL could be performed 
and that these TIL could be infused with no exposure of the patient to a 
replication competent virus. These studies have provided us with valuable 
experience to perform the proposed studies with TNF modified TIL with greater 
ease and safety. 
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Recombinant DNA Research, Volume 14 
