established from a patient, AV, in 1975 from regional lymph node 
metastases (see reference 29). Patient AV originally was diagnosed with 
melanoma Clark’s level IV of the anterior chest wall in 1974 at age 18. In 
March 1975, melanoma recurred in multiple left axillary lymph nodes 
with deep invasion into soft tissue. This disease was resected and he 
received experimental adjuvant immunotherapy with Corynebacteria 
parvum. In August 1975, disease recurred in the left supraclavicular and 
infraclavicular area. He had surgical resection and started vaccine 
treatment with irradiated autologous melanoma cells and BCG adjuvant. 
In December 1975, the patient had a third recurrence of tumors in the left 
neck with invasion of the axillary vein and brachial plexus. He was 
treated with surgical resection followed by radiation therapy, then 
dacarbazine adjuvant chemotherapy in 1976-1977. In 1977, he restarted 
vaccination with autologous tumor cells and BCG. He developed a 
recurrent melanoma in the left neck in February 1978 that was resected. 
The vaccine was continued up to 1980. As of 1991, he has had no further 
recurrence of melanoma. CD8-positive T cells were derived from 
peripheral blood after vaccination that efficiently lysed autologous SK- 
MEL-29 tumor cells in an HLA-A2 restricted fashion, and have been 
extensively reported by Knuth and colleagues (52,56). 
The SK-MEL-29 melanoma cell line has been infected with the retroviral 
vector, NAP-AD/IL-2 (see Appendix C for construction). Southern 
blotting demonstrated appropriate integration of the provirus into the host 
chromosomes. Additional studies revealed this cell line to be mycoplasma 
and helper virus free. Those tumor cells constitutively and stably secrete 
40 U IL-2/ml. Cells will be irradiated with 10,000 rads prior to injection, 
to assure their inability to proliferate (see appendix F). Two dose levels of 
melanoma cells will be used for the vaccines. 
2.9 Clinical Protocol 
This is a pilot study of immunization with SK-MEL-29 allogeneic melanoma 
cells that have been altered to secrete IL-2, given to patients with stage IV 
metastatic melanoma, in order to test the safety of this procedure. Patients will 
be HLA tissue typed. Patients who are HLA-A2 positive will receive 
vaccinations of allogenic SK-MEL-29 cells that secrete IL-2. The expected 
accrual for this study is twelve patients. The expected duration for the trial will 
be 6-12 months. 
Tissue culture of SK-MEL-29 cells will be grown in the laboratories of Drs. 
Bemd Gansbacher and Alan Houghton. The melanoma cells have been infected 
with the retroviral vector NAP-AD/IL2. Cells that secrete IL-2 will be irradiated 
(10,000 rad) and given as a series of four vaccinations. If a patient has a 
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