can develop microscopic metastatic implantation, but these studies show that 
these implants fail to reach a significant size prior to the patient’s death 
from their pre-existing known metastatic disease. Certainly the cells which 
might escape from a small, cutaneously-implanted tumor site into the systemic 
circulation (if this occurs at all), would be far less than that from the 
intravenous auto-inoculation which occurs on a daily basis in these shunted 
patients or in any patient with widely metastatic cancer. 
Thus, data suggests that auto-inoculation of a small amount of viable 
tumor at an isolated cutaneous site will often generate tumors for TIL growth, 
but that in the setting of widely metastatic cancer, such an approach is 
unlikely to significantly affect survival or the disease course of such 
patients. In the experiments in this protocol, however, the introduction of 
the TNF gene into tumor is designed to increase the immunogenic ity of the 
tumor (as reviewed earlier) and will prevent tumor growth in most cases. The 
use of draining lymph node lymphocytes can thus provide a source of cells for 
use in adoptive immunotherapy of these patients. 
Shu et al. have published data in murine models showing that draining 
lymph node lymphocytes (DLNL) from sites of tumor immunization can be 
sensitized in vitro (by mixed tumor-lymphocyte culture) and expanded in IL-2 
(32-34). These cells can then be adoptively transferred to tumor-bearing mice 
and show antitumor activity (Table 7). Many of the features of these cells 
are similar to TIL (such as phenotype and tumor specificity patterns) although 
they have a lesser capability to expand in vitro and may be somewhat less 
effective on a cell-for-cell basis (25,35). These cultured draining lymph 
node cells are currently undergoing Phase I testing by Dr. Alfred Chang at the 
University of Michigan. In order to provide patients undergoing tumor 
immunization with a treatment alternative in the event of failure of the 
primary TIL culture, at the time of resection of inoculation sites (or at 
three weeks after inoculation if no tumor growth is apparent), draining lymph 
nodes will be excisionally biopsied to prepare an alternative T-cell culture 
for adoptive transfer. This will be performed with sensitization using 
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Recombinant DNA Research, Volume 15 
