Recombinant DMA Advisory Committee - 5/30-31/91 
IV. PROPOSED ADDITION TO APPENDIX D OF THE NIH GUIDELINES REGARDING A 
HUMAN GENE TRANSFER PROTOCOL ENTITLED THE ADMINISTRATION OF 
INTERLEUKIN‘2, INTERLEUKIN-4, AND TUMOR INFILTRATING LYMPHOCYTES TO 
PATIENTS WITH MELANOMA: 
Dr. Lotze said that because this protocol had been before both the HGTS and the RAC on 
previous occasions he would only discuss issues which had arisen at the previous RAC meeting 
and then would ask the primary reviewers to comment on the protocol. 
He said that the Anderson-Blaese-Rosenberg protocol had shown that it was possible to 
introduce the neomycin-resistance gene into patients via the tumor infiltrating lymphocyte 
(TIL) and that both transduced and non-transduced cells can be grown which appear to have 
the desired biologic characteristic of G418 sensitivity. Furthermore, these cells can be 
detected in peripheral blood out to at least 2-3 weeks and in the tumor out as far as a couple 
of months. 
Dr. Lotze said that the only major difference between the Anderson-Blaese-Rosenberg 
protocol and the one he was proposing was the incorporation of interleukin-4, another T cell 
growth factor, and that in every other respect this protocol is identical to that which was 
approved by the RAC for the National Cancer Institute (NCI). 
Dr. Lotze said that his laboratory had demonstrated their ability to grow cells from human 
tumors in combinations of IL-2 and IL-4 and that cells so cultured can indeed be marked with 
the neo gene using the retroviral vectors. In addition, he noted that additional studies had 
been undertaken to expand over 15 different TIL preparations with combinations of 11^2 and 
IL-4 and cells had been transduced and/or selected. In all cases negative controls were used 
to look for neo expression and no such expression was found, however in situations in which 
the cells were selected for neo expression and grown in IL-2/IL-4 there was increased 
expression of neo. 
Dr. Lotze underlined the fact that the company which will perform the PCR assays for them 
is capable of detecting down to 1 in 10^ cells and that the plan is to have PCR done on 
clinical specimens by that company. 
Dr. Lotze noted that one question that had arisen dealt with quantitation of lymphocytes in 
individual tissues. He said that after consideration of many techniques for accomplishing this, 
it was felt that standard immunohistochemistry would afford the most straightforward method 
of assessing this parameter. He presented a series of slides showing the ability of standard 
immunohistochemistry to quantitate lymphocytes in various tissue samples. 
In conclusion. Dr. Lotze reminded the committee that this protocol had been through 
extensive review and modification and that due to this process many typographical errors had 
been made due to the multitude of changes that have been requested in the protocol. He 
noted that the investigators were anxious to begin the studies and hopeful of receiving RAC 
approval to proceed. He underlined that official approval is still needed from the Cancer 
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Recombinant DNA Research, Volume 14 
