cells as well as normal tumor cells mixed at the same site. In this proposal 
we thus plan to take advantage of the increased immunogenic ity of these TNF 
producing tumors to attempt to immunize patients with advanced and otherwise 
untreatable metastatic cancer. These tumor cells will be injected 
subcutaneously and intradermally into the thigh in an area that can be 
followed easily in an attempt to both immunize the patient against their 
cancer as well as to provide local immunization that could be used to grow 
lymphocytes for use in adoptive therapy. Lymphocytes will be obtained either 
from the draining inguinal lymph node group or from the tumor site itself. 
In addition to the increased immunogenicity of the gene modified tumor, 
recent work in the Surgery Branch has suggested that the subcutaneous 
injection of tumor can lead to the development of more effective tumor 
infiltrating lymphocytes. In 12 successive experiments, TIL grown from 
visceral sites were simultaneously tested by careful in vivo titration against 
TIL from tumor injected into the cutaneous site. In 11 of 12 experiments, TIL 
from the subcutaneous location were more effective than those at visceral 
sites (see Table 6). In three other experiments, TIL from murine tumors in 
the liver were less effective than TIL from cutaneous sites. 
In the human, other workers have shown that small cutaneous tumor auto 
inoculations can provoke significant immune responses. Hoover and Hanna have 
published work in which colorectal cancers from primary sites were irradiated 
and utilized for autologous immunization of patients in the adjuvant setting 
(22). Patients with Dukes B2 and C tumors demonstrated improved overall 
survival in a randomized study, presumably due to an effective immune response 
to the immunization. In addition, Berg and Mastrangelo have investigated the 
immunization of melanoma patients with irradiated autologous tumor, and have 
demonstrated the induction of significant T-cell infiltrates in tumors as well 
as rare clinical responses in patients with metastatic disease (23,24). These 
studies generally utilized irradiated tumor cell inoculation and therefore 
result in no tumor for the generation of TIL. The presence of tumor is vital 
for generating optimal TIL in that T-cells separated from fresh tumors will 
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Recombinant DNA Research, Volume 15 
