I. Brief scientific abstract 
A tumor nodule will be resected from a patient with advanced -Cancer and 
the lymphocytes (TIL) infiltrating into this cancer deposit will be cultured 
and expanded in vitro using techniques previously used in our adoptive immuno- 
therapy protocols. tJhen these TIL have reached log phase growth they will be 
incubated with a retroviral vector LTSN (containing genes for tumor necrosis 
factor and for neomycin resistance). These cells will then be expanded in 
culture until approximately 4 x 10^^ cells are achieved. These TIL will then be 
cr yopreserved in 10^*^ cell aliquots and tested to insure that they are free of 
replication competent virus and to determine the levels of tumor necrosis 
factor that are secreted. These gene-transduced TIL will then be administered 
to the cancer patient in escalating doses beginning at 10^^ TIL along with 
180,000 lU/kg of IL-2 every 3 hours for up to five days. Three weeks later the 
dose will be escalated to 3 x 10^*^ TIL and at three weekly intervals, escalated 
to 10^^ and then to 3 x 10^^ TIL. Extensive monitoring of patients will be performed 
to test for any toxic side effects resulting from the TIL administration. 
Studies of the survival and distribution of the gene-modified TIL will be 
performed by sequential sampling of blood, lymph nodes and tumor biopsy material. 
Tests for in vivo production of tumor necrosis factor will be conducted by 
measuring serum TNF levels and tests for the presence of the TNF gene and the 
NeoR gene will be performed by PCR-DNA analysis. Evaluation of the possible 
therapeutic response of cancer deposits to this therapy will be measured. The 
maximum dose of gene-transduced TIL that can be safely administered, will be 
determined in at least five patients. In subsequent patients transduced TIL 
will be selected in G418 and a similar escalating dose protocol will begin 
starting at 10% of the established safe number of unselected cells. When safe 
levels of non-selected and selected gene transduced TIL are determined, the 
dose of IL-2 will be raised to 720,000 lU/kg in a subsequent series of patients. 
Recombinant DNA Research, Volume 14 
[305] 
