July 30, 1990, Human Gene Therapy Subcommittee 
TILs making more TNF and cleared via the same mechanisms without adverse 
consequences. LAK cells, which make 500-600 picograms of TNF per 10*^ cells, have 
been given to approximately 200 patients, and they are cleared by the liver and spleen in 
an identical fashion to the method of clearance of TIL cells. No adverse organ-specific 
effects have been noted. 
Dr. Rosenberg said that after the proposal had been turned down by the NCI Clinical 
Research Committee, the NHLBI Clinical Research Committee, and NIH IBC, 
additional data were gathered and some technical changes in the protocol were made 
which resulted in their approving the protocol. The result of this was the addendum to 
the protocol that was supplied to the HGTS. The IBC had approved the protocol with 
three stipulations: 
1. That evidence be presented to show that TNF production by the tumor 
cells was the factor that led to their regression; 
2. That studies be performed in an animal model to show that tumor 
regression could be abrogated by use of anti-TNF antibody; and, 
3. That safety studies be performed in a primate model, along with 
presentation of additional information based on longer follow-up of the 
N2/TIL protocol. 
Dr. Rosenberg noted that Dr. Carter, Chairman of the NIH IBC, was in attendance at 
today's meeting and that new information would be presented regarding these three 
points. 
Dr. Rosenberg said that, with regard to evidence showing that TNF production by tumor 
cells was the factor leading to their regression, a study was performed by Dr. Anthony 
Asher and Jim Mule, of his laboratory, in which animals received tumor cells that were 
either transduced with a TNF-neo vector or just a neo vector. The animals given the 
TNF-neo vector showed tumor growth and then regression. These animals were then 
treated with either a control antibody or anti-TNF antibody. The anti-TNF antibody had 
no impact on the neo-producing cells alone. However, animals that had the neo-TNF 
vector, who received anti-TNF antibody, had tumor growth. 
He also presented results of studies of the N2/TIL protocol in which data has been 
gathered on eight patients to date, three of which have died of progressive melanoma. 
After examining over 25 samples obtained from autopsies of two of the patients, 
sampling from multiple sites, no positive gene modified cells were detectable, even at the 
1 in 100,000 level. There is one positive sample which is being re-evaluated, as it is 
suspected of being a false-positive. 
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