were injected ip into BNX mice. 9 animals were reconstituted with this procedure, six with 
transduced cells and three with non-tranduced ADA(-)PBL as negative controls. Twenty BNX 
mice were reconstituted with normal PBL as positive controls for the efficiency of this human- 
mouse model. In vivo long term survival of human cells in the recipient BNX mice was 
demonstrated in the spleen by DNA analysis and the spleen lysates showed the typical band of 
human ADA by Cellogel analysis. Four weeks after reconstitution, human T and B cells were 
detected in the peripheral blood and spleen by FACS analysis. 
Immune function of the transferred human cells was evaluated by testing for the presence of 
human IgG in the mouse blood and measurement of human alloreactive T cells in the spleens of 
the recipient BNX mice. The mice reconstituted with hADA transduced ADA(-)lymphocytes 
produced human IgG at levels comparable to BNX mice reconstituted with normal PBL. By 
contrast, no human IgG was detected in the blood of recipient mice reconstituted with the non- 
transduced ADA(-)lymphocytes, even though BNX mice have normal amounts of ADA in their 
own cells and do not have elevated levels of circulating or tissue deoxyadenosine. Allospecific 
human T cell clones were recovered from the spleens of BNX recipients reconstituted with 
transduced PBL, but not non-transduced PBL, and the recovered T cells expressed vector 
derived hADA. These observations suggest that intracellular correction of ADA(-)lymphocytes 
may be more beneficial for immune reconstitution than extracellular detoxification alone. 
1 .5.4 In Vivo Human Experiments with TIL Transduced with the LNL6 Vector 
On January 19, 1989, Dr. James Wyngaarden approved our human gene transfer N2/TIL 
clinical protocol (protocol 86-C-183c). At present, 8 patients with advanced malignant 
melanoma have received tumor infiltrating lymphocytes (TIL) that have been marked with the 
safety-modified N2 vector called LNL6. The first 5 patients were treated between May 22 and 
July 21, 1989. Data from these first five patients has been prepared for publication and a 
preprint of the manuscript has been made available. 
Because we realized that this initial human gene transfer clinical protocol would provide a basis 
for risk assessment for future human gene therapy protocols, we carried out extensive safety 
studies on; a) the retroviral supernatant prior to transduction of human TIL, b) the transduced 
TIL before they were infused into the patient, and c) the patients who received the gene-marked 
TIL. 
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