Recombinant DNA Advisory Committee - 10/7-8/91 
Dr. Leventhal said she would discuss the TNF protocol. In this experiment. Dr. 
Rosenberg plans to remove tumor from the patients that present with a diagnosis of 
melanoma, renal cell carcinoma, or colon cancer. He will attempt to grow these tumor 
cells in culture and, if successful, transduce these lines with the TNF gene. If the 
patients should relapse or their cancer progresses, these transduced cells will be injected 
subcutaneously into the thigh, and draining lymph nodes will be harvested 21 days later. 
Lymphocytes from these draining lymph nodes will be expanded and then reinfused into 
the patients along with IL-2. She pointed out that cell lines are cultivated for every 
patient who undergoes surgery at the NIH for primary therapy. These tumor cells will 
be cryopreserved and will be administered to the patient should he/she have recurrent 
tumor. Dr. Leventhal expressed concern that there would not be time, in a patient's 
rapidly progressing disease, to perform all of the tasks in the protocol. 
Dr. Leventhal said the rationale for these experiments is that the TNF transduced cells 
are more immunogenic than untransduced cells. In the model experiments, the modified 
cells do not appear to grow as well as the unmodified cells, but an immunologic basis for 
this growth disparity is not completely proven. In Dr. Rosenberg's previous study with 
neo-modified TIL, he showed that the TIL home to tumor in three of five patients. 
However, it is not clear how many of the cells that are removed from the lymph node, 
after subcutaneous injection of modified tumor cells, can be called TIL or TIL 
precursors. Dr. Leventhal questioned whether the tumor cells will grow in the patient 
after they are modified. If they do grow, they may endanger the patient because they are 
seeded with cells that make TNF constantly. This may render the patient symptomatic 
as TNF causes chills and fever. If the cells grow and remain in the patient forever, there 
would be the question of chronic toxicity. If the cells do not grow, will they be capable 
of stimulating the lymphocytes in the draining lymph nodes? 
Dr. Leventhal recounted that due to these concerns the HGTS elected to approve the 
initial treatment of five patients with the stipulation that the local reaction be observed 
closely, and that measurements, descriptions, or photographs of the injection sites be 
submitted. The draining lymph nodes and other tumor sites should be studied by 
polymerase chain reaction (PCR) for signs of spreading transduced tumor cells and to 
study the trafficking pattern. The cells harvested from the lymph nodes should be 
studied to determine the cell types present. A toxicity reporting scale should be 
developed that provides more detail about the use of IL-2 and reinfused cells. She noted 
that Dr. Rosenberg had agreed to limit his initial treatment with this protocol to five 
patients and to perform PCR analysis of the draining lymph nodes near the cell injection 
site. 
Dr. Haselkom asked the investigators to discuss why the TNF-modified cells regress in 
the nude mouse. He asked if any experiments had been performed with animals that 
have established tumors rather than animals that have had tumor cells injected. With 
Recombinant DNA Research, Volume 15 
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