HGTS 7/29/88 
At this meeting of the Subcommittee Drs. Anderson, Lotze, and Blaese presented their 
human gene transfer protocol. Dr. Anderson began with an overview of the proposal with 
the rational for using TILs, Dr. Lotze discussed the use of adoptive immunotherapy in the 
treatment of cancer and particular the treatment of malignant melanoma; Dr. Blaese 
commented on the marking procedure for the lymphocytes and the advantages of 
genetic labeling. Dr Anderson returned to discuss safety issues in the proposal. 
A scientific summary of the protocol is as follows: 
Tumor infiltrating lymphocytes (TIL) would be isolated from a patient’s tumor 
and grown in culture in the presence of interleukin-2 (IL-2). An aliquot of 
TIL would be removed at the time they reached log phase growth. The 
aliquot (representing no more than one-third of the total TIL population) 
would be incubated with the retroviral vector N2 (containing the gene 
coding for neomycin resistance, or Neo R ). This treated aliquot would be 
grown in media containing G418, a neomycin analog in which only those 
cells expressing the Neo R gene can survive. The cells would be tested to 
insure that they are virus-free, have similar surface antigen patterns to the 
parent TIL population, and have not changed significantly in other 
properties (includir g continued dependence on exogenous IL-2 for growth). 
The treated aliquot would then be administered to the patient along with the 
bulk TIL population that would have been grown separately. The proportion 
of marked TIL in the final TIL population that would be returned to the 
patient would be between 5-30%. After administration, samples of blood, 
lymph nodes, and tumor biopsy material (already being obtained as part of 
the standard TIL protocol) would be tested for the presence of the Neo R 
gene by PCR DNA analysis. The marked TIL would be recovered by 
growth of the tissue sample in IL-2 medium plus G418. The recovered cells 
would be studied for phenotypic and cytotoxic properties in order to 
attempt to learn why TIL immunotherapy is successful in some cases but 
not in others. 
The Subcommittee wrestled with the issue of whether the protocol was, in reality, human 
gene therapy and whether it fell under the jurisdiction of the Recombinant DNA Advisory 
Committee and its Subcommittee. After deliberation, it was decided the protocol was 
"very similar to human gene therapy," and that the Subcommittee should review it. 
Recombinant DNA Research, Volume 13 
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