tumor necrosis factor will be inserted into these tumor cells using r ather 
retroviral mediated gene transduction -o r by t r ansfection Let ' hlligu u a uaing- 
&a - Ls-iru m - phos p ha t e or microin j option ' . Patients that develop advanced 
u n treatable metastatic cancer and have failed all other effective treatm e nt s 
will be offered the pos sibility of joining this protocol. After signing an 
informed consent the patients will be regi stered in the protocol and wi ll 
receive the injection of the gene modified tumor cells into the mid thigh . Up 
to 2 x 10 8 gene modified tumor cells will be injected in 1 ml subcutaneously 
and three centimeters lateral or vertical from this site the patient will 
receive two intradermal injections of 2 x 10 7 gene modified tumor cells in 0.1 
ml each. These sites will then be carefully monitored. At three weeks the 
patient will undergo resection of several lymph nodes from the 
draining superficial inquinal area. These lymph nodes will be used to grow 
lymphocytes for adoptive cellular therapy of that patient. As predicted by 
animal models, it is not expected that tumor will grow at the local injection 
site. If, however, tumor. does grow then the soft tissue in the area of the 
\ 
tumor injection will be resected when a tumor has reached one to two 
centimeters and an attempt will be made to grow tumor infiltrating lymphocytes 
from this site. Lymphocytes either from draining lymph nodes or from the 
tumor site itself (whichever become available first) will be grown in vitro by 
the standard techniques used in many previous protocols and assays published 
detail. These lymphocytes will then be adoptively transferred to the patient 
along with 720,000 IU/kg of IL-2, exactly as in our previous protocols and as 
previously published in detail. The impact of the immunization procedure on 
established tumor at other sites will also be monitored to evaluate whether 
this immunization procedure itself has anti-tumor effects. Biopsies of 
cutaneous or subcutaneous lesions may be performed. 
2 . Preparation of the TNF-NeoR vector containing supernatant. The TNF- 
NeoR vector used in this protocol is identical to that used in our approved 
protocol (#90-C-186) utilizing the introduction of these genes into human 
tumor infiltrating lymphocytes for use in human therapy. The following 
[ 68 ] 
Recombinant DNA Research, Volume 15 
