SCIENTIFIC ABSTRACT 
A Phase I Study of Vaccination with Autologous, Irradiated Melanoma Cells 
Engineered to Secrete Human Granulocyte-Macrophage Colony Stimulating 
Factor 
This clinical trial for patients with metastatic melanoma will investigate the 
use as therapeutic vaccines of autologous, irradiated melanoma cells 
engineered by retroviral mediated gene transfer to secrete human 
granulocyte-macrophage colony stimulating factor (GM-CSF). A total of 16-25 
patients will be treated at three different dose levels of vaccine. Each patient 
will receive inoculations of 1x10^ autologous melanoma cells (secreting 40- 
1000 ng of GM-CSF/IO^ cells/24 hours) subcutaneously and intradermally. The 
interval between vaccinations will be varied from every month to every two 
weeks to weekly for a total duration of three months. This design will test the 
effects of increasing both the frequency of vaccination and the total number 
of cells and GM-CSF administered. 
The proposed study is based on pre-clinical experiments in murine tumor 
model systems which indicated that injection of irradiated tumor cells 
engineered to secrete murine granulocyte-macrophage colony stimulating 
factor generated potent, specific, and long lasting anti-tumor immunity. 
Efficacy of irradiated, GM-CSF expressing cells could be demonstrated in models 
of melanoma, renal cell carcinoma, fibrosarcoma, lung carcinoma, colon 
carcinoma, neuroblastoma, bladder carcinoma, and prostate carcinoma. 
The MFG-S replication defective retroviral vector provides efficient transfer 
and expression of the gene encoding human granulocyte-macrophage colony 
stimulating factor in primary short term melanoma explants. As no selection 
for transduced cells is required, antigenic heterogeneity within the 
vaccinating inoculum is relatively preserved. The vaccinating cells are 
lethal ly irradiated with 15,000 rads (which does not interfere with their GM- 
CSF production) to provide safeguards against both the injection of tumor cells 
potentially rendered more virulent by in vitro manipulation or insertional 
mutagenesis, and the autonomous growth of nonneoplastic cells induced by 
autocrine synthesis of their own growth factors. 
The overall goals of the proposed phase I study are: 
1. To evaluate the safety of clinical administration of autologous, 
irradiated melanoma cells engineered, by retroviral mediated gene 
transfer, to secrete human granulocyte-macrophage colony stimulating 
factor. 
2. To determine, if possible, the dose limiting toxicity (DLT) and 
maximum tolerated dose (MTD) of autologous, irradiated melanoma 
cells engineered to express GM-CSF. 
3. To describe and quantify any local or systemic immune response 
stimulated by vaccination with autologous, irradiated GM-CSF 
expressing cells. 
Recombinant DNA Research, Volume 20 
