Human Gene Therapy Subcommittee - 11/21-22/91 
Dr. Economou explained that two different retroviral vectors would be used to transduce 
TIL and PBL; these vectors are referred to as LNL6 and GINa. The major differences 
between these two vectors is the insertion of a small polylinker sequence to allow them 
to be distinguishable by PCR. Data were presented showing that purified and isolated 
CD8( + ) TEL and CD4( + ) TIL could be transduced with these vectors and that there was 
appropriate integration with the proper primers. The data also indicated that a single 
gene copy of LNL6 or GINa could be detected in 10 5 cells using these primers. 
This protocol would allow the investigators to support or refute the hypothesis that TIL 
have specific localizing properties. However, the ratio of TIL to PBL being infused must 
be determined. At various intervals, the investigators will sample blood and biopsy 
tumor, uninvolved skin, and skeletal muscle to compare these ratios. 
Dr. Economou emphasized that biopsy strategy is critical since the procedure will 
probably not directly benefit the patient. Inclusion criteria for entering into this protocol 
require that patients have subcutaneous disease. The reason for this requirement is that 
these subcutaneous nodules can be biopsied at various intervals. A diamond block 
anesthetic would be placed around a small tumor deposit to perform an elliptical 
incision. The tumor would then be removed, the adjacent fat, skin at the corners of the 
incision, and then the fascia would be opened up to remove a couple of fascicles of 
skeletal muscle. Therefore, only one anesthetic would be required to obtain four 
different tissues. These specimens would be used for pathology analysis and semi- 
quantitative PCR. 
The protocol involves obtaining immunological information on these TILs. 
Cryopreserved autologous tumor specimens will be available to test the specificity of TIL 
compared to allogeneic tumor cell lines. Autologous tumor cell lines can be generated 
in approximately 80% of these patients. TIL will be analyzed for gene expression of a 
number of cytokines and for protein production. Hopefully, the following scientific 
information will be yielded from this protocol: (1) a definitive and unambiguous test of 
the hypothesis that TIL will localize to tumor, (2) the ability to compare bulk TIL versus 
CD8( + ) and CD4( + ) TIL in vivo trafficking, and (3) a correlation of in vivo behavior 
with cytotoxicity, phenotype, and cytokine production. 
Dr. Economou introduced his co-investigator, Dr. R. E. Belldegrun from the Division of 
Urology at UCLA. Dr. Belldegrun will be responsible for the urologic cancer patients. 
Presentation-Dr. Belldegrun 
Dr. Belldegrun said that unlike melanoma, renal TIL are a heterogeneous population of 
cells. These TIL exhibit a short period of lytic activity, approximately four to five weeks, 
and then disappear. The phenotype of renal TIL differs from melanoma; it corresponds 
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