2.0 STUDY DESIGN 
This is phase I, single site, dose escalating study using fresh or cultured autologous melanoma 
admixed with IL-2 producing allogeneic melanoma cell line M-24 and administered as an 
irradiated vaccine. The choice of fresh or cultured autologous tumor cells will depend on the 
ability to establish a cell line and the amount of tumor resected. Patients will be vaccinated on 
a biweekly basis for 3 months, then monthly thereafter for up to one year. Eligible patients will 
have metastatic melanoma. Patients will be followed and studied bi-weekly for evidence of a 
tumor-specific immunological or clinical response. The groups of 10 patients will receive 
increasing numbers of IL-2 transduced M-24 with a constant number of tumor cells as a dose 
escalation to evaluate toxicity and efficacy. 
Vaccine composition groups will be as follows: 
Group A 
Groun B 
GrouD C 
Autologous melanoma 
(fresh tumor or cultured 
cell lines) 
10 7 
10 7 
10 7 
Addition of sufficient numbers 
of IL-2 transduced M-24 
to produce the following amount 
of IL-2 per 24 hr. 
KPpg 
lCfpg 
10 5 pg 
Number of Patients Treated 
10 
10 
10 
3.0 STUDY TREATMENT 
PREPARATION OF GENETICALLY-ENGINEERED AUTOLOGOUS TUMOR CELL 
VACCINE 
All manipulations will be conducted in the Cellular and Genetic Therapy Core 
Laboratory in the UCLA Jonsson Comprehensive Cancer Center. 
3.1 Preparation of Autologous Tumor 
Patient vaccines will be composed of either freshly cryopreserved autologous 
tumor or tissue culture cell lines established from the patient’s tumor. 
3.1.1. Preparation of Tumor Cells 
Solid tumors will be resected in the operating room, minced into 5 mm 3 pieces 
and placed in an enzymatic digestion medium composed of RPMI 1640 (Flow 
Laboratories), containing hyaluronidase type V (0.01%), DNase I (0.002%), 
collagenase type IV (0.01%). (Sigma, St. Louis, MO), penicillin (50 IU/ml), 
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