tumor and patient comfort, but an effort will be made to obtain samples at weekly or biweekly 
intervals. Peripheral blood draws will be coordinated with these biopsies. These samples will 
be subjected to semiquantitative PCR analysis. See Appendix XI for biopsy schedule. 
(c) AUTOPSY - Should a treated patient expire within 6 months of TIL infusion, 
permission for autopsy will be requested and samples of brain, lung, liver, spleen, 
intestine, kidney, skeletal muscle, skin, blood and tumor obtained. These will be 
subjected to semi-quantitative PCR analysis. 
(d) RESEARCH GOALS - Although TIL appear to participate in the regression 
of certain tumors, their mechanism of action isn’t understood. TIL may 
specifically localize to tumor sites and mediate tumor destruction, or, 
alternatively, may help to generate an effective systemic immune response without 
tumor localization. Animal models of adoptive immunotherapy have demonstrated 
that both CD8 and CD4 immune cells are able to mediate tumor destruction. 
In designing this clinical trial, we wished to test the hypothesis that TIL have 
unique immunological and in vivo trafficking properties . We have elected to 
compare bulk TIL, CD8(+)TIL and CD4(+)TIL with respect to in vivo life-span 
and trafficking. If TIL have "special" properties, we would expect that they 
would behave differendy than IL-2 expanded bulk, CD8(+) or CD4( + )PBL. 
Accordingly, we have selected PBL as a "control" effector cell. In addition to 
having bulk, CD8 and CD4 TIL expanded, PBL will be obtained, enriched and 
expanded in parallel cultures. We will use PCR distinguishable neoR retroviral 
vectors to label the 2 different effector populations. Using semiquantitative PCR, 
we will know the rado of administered cell populations (e.g. bulk TIL/PBL, CD8 
TIL/CD8 PBL, CD4 TIL/CD4 PBL, CD8 TIL/CD4 TIL). This ratio can then 
be determined in samples of blood, tumor and normal tissue obtained at intervals 
following adoptive transfer. If the ratio changes significantiy and in a 
reproducible fashion, we can conclude that the in vivo trafficking properties of 
these cells differ. We would be particularly interested to see if TIL populations 
are enriched in tumor deposits and whether this is a result of localization or 
improved life-span. 
16. STATISTICAL CONSIDERATIONS 
Study subjects will be considered responders to the combination of TIL and rIL-2, and 
IFNaA if, by the end of the observation period they have achieved an objective response 
(either a complete response or partial response) of at least 4 weeks duration. The 
objective response rate will be determined as the percentage of all evaluable study 
subjects considered as responders to the described therapy. 
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Recombinant DNA Research, Volume 15 
