TIL will be grown from patients and will be transduced with the TNF-NeoR 
retroviral vector and expanded and cryopreserved in at least four aliquots con- 
taining a minimum of 10^*^ TIL. Extensive safety and functional tests will be 
conducted on these TIL during this time. For infusion these TIL will be thawed 
and re-expanded for a short time (1 to 3 weeks) in culture prior to reinfuslon. 
Patients will first receive a single infusion of 10^^ TNF-modified TIL along 
with the Infusion of IL-2 at 180,000 lU/kg every eight hours for up to 5 days. 
This dose of IL-2 is the equivalent of 30,000 Cetus units/Kg which is approximately 
one-third the dose of IL-2 used in our previous protocol using NeoR gene modified 
TIL (86-C-183c). If patients do not reach Grade IV toxicity (see Appendix B) 
or if they reach Grade IV toxicity easily managed by concomitant medications 
(as in previous IL-2 or TNF protocols) then patients will be escalated three 
weeks later to receive 3 x 10^^ TIL plus IL-2 in a similar fashion. If tolerated, 
three weeks later they will receive 10^^ TIL plus IL-2 in a similar fashion. 
If tolerated, three weeks later, 2-3 x 10^^ TIL will be administered. At least 
five patients will be treated in this fashion. When the maximum dose of TNF- 
modified TIL is established (or the 2-3 x 10^^ TIL dose is achieved without 
Grade IV toxicity as mentioned above) a second series of patients will receive 
TNF modified TIL selected in G418 using a similar escalating schedule but 
starting at 1/lOth the dose of unselected TIL found acceptable for patient 
infusion. The maximum dose of TNF-modified TIL will be determined as the TNF 
TIL dose tolerated by at least 80% of patients. Detailed studies of patient 
toxicity as well as possible therapeutic effects will be noted. Detailed 
studies of the survival and distribution of these cells in patients will be 
conducted. Safety and monitoring studies will be performed In a fashion 
virtually identical to those in our previously approved protocol for the use 
of TIL modified by the LNL6 retroviral vector (86-C-183c; reference 21, 
attached) . 
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Recombinant DNA Research, Volume 14 
