SCIENTIFIC ABSTRACT 
Scientific Abstract 
Although it has been well established experimentally that the transfer of sensitized T 
lymphocytes can mediate potent antitumor effects, extrapolating the principles of adoptive 
immunotherapy obtained from animal studies to clinical therapy will require the development 
of innovative techniques to isolate and propagate antitumor effector T cells from cancer 
patients. Toward this end, we have established culture methods whereby cells from tumor- 
draining or tumor primed lymph nodes (LN) can be sensitized to acquire therapeutic efficacy. 
Since these cells do not demonstrate overt antitumor reactivity before culture, they are 
functionally referred to as "pre-effector" cells. One method to generate antitumor effector T 
cells involves the sequential activation of pre-effector LN cells with anti-CD3 mAb followed by 
expansion in low concentrations of IL-2. Animal experiments have demonstrated that the 
antitumor reactivity of these anti-CD3/IL-2 activated cells are exquisitely tumor-specific and 
mediate the regression of established tumor in adoptive immunotherapy. 
A major obstacle which confronts the clinical application of adoptive immunotherapy is 
the relatively weak immunogenicity of human cancers which hampers the induction of 
sensitized pre-effector cells. Recent observations in animal studies indicate the tumors can 
be genetically altered to enhance the host immune response against native or parental tumor 
antigens. The transfection of tumor cells with some cytokine genes have often resulted in T 
cell mediated rejection of these tumors by their syngeneic hosts. Similarly, we found that the 
transfection of the poorly immunogenic B16BL6 murine melanoma tumor with the IL-4 gene 
resulted in decreased tumorigenicity when inoculated into the syngeneic host. More 
importantly, draining LN cells removed from these animals and activated by the anti-CD3/IL-2 
culture procedure generated potent therapeutic effector cells which mediated the adoptive 
immunotherapy of established metastatic parental tumors. These observations provide the 
rationale for this clinical protocol to examine autologous tumor cells modified with the IL-4 
gene, which will be utilized as a vaccine to induce pre-effector LN cells in patients with 
metastatic melanoma. These vaccine-primed LN cells will be activated by the anti-CD3/IL-2 
method and subsequently transferred intravenously to patients along with the concomitant 
administration of IL-2 (360,000 lU/kg q8h x 5 days) to support their survival/function in vivo. 
The specific aims of the protocol are: 1 ) To assess the feasibility and toxicity of 
adoptive T cell immunotherapy of melanoma with anti-CD3/IL-2 activated LN cells that are 
primed in vivo with IL-4 modified autologous tumor cells, 2) To evaluate the antitumor 
efficacy and in vivo immunological reactivity of patients receiving adoptively transferred T 
cells, and 3) To investigate the in vitro immunological reactivities of the activated T cells that 
might correlate with their in vivo antitumor function. 
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Recombinant DNA Research, Volume 18 
