Human Gene Therapy Subcommittee - 7/30/90 
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. 
Dr. Rosenberg then presented data on monkeys that had received 
human TILs transduced by the exact vectors to be used in humans, 
following a 20 mg/kg dose of cyclophosphamide. Two monkeys 
received no cells, two received 5 X 10' cells per kg of neo- 
vector TIL, two received TIL^jjp selected cells producing 1,942 
picograms per 10® cells per 24 hours, and two monkeys received 
non-selected TIL,pj,p in a concentration equal to the that proposed 
for infusion into humans based on body weight. The monkeys (two 
groups) have been followed up from 7 to 30 days, and no adverse 
effects and no toxicity have been seen in any of the monkeys. 
Dr. Rosenberg added that the supernatant will be tested for virus 
using the S'''/L*’ assay and some more sensitive assays; Western 
blots will be performed on patient sera. With respect to long- 
term effects of IL-2, cells stop circulating and toxicity ceases 
when IL-2 is stopped, although in two patients there was survival 
of a tiny number of cells out to two months post cessation of IL- 
2 . 
Dr. Rosenberg addressed the issue of transduced cells being able 
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