Table 7 
Treatment of B16 melanoma 
cell*. 
lung mctajtaics with Irradiated CM-CSF 
traniduced tumor 
Treatment 
mean / lung tumori/mouse 
#mlce tumor free 
Exp 1* 
Exp 2 b 
Exp 2 b 
None 
32 
. 1 I.S 
0/10 
Irradiated wild type 
32 
S.J 
0/10 
Irradiated GM-CSF 
12 
0.2 
0/10 
* C378L6 mice were Injected In the tall vein with 2xl0 4 live wild type B 1 6 melanoma 
eellj. 3 dayi later they were Infected lubcutaneouily with irradiated wild type or 
Irradiated GM-CSF transduced B16 melanoma cells. 28 days later, lungj were removed 
and examined both microscopically and microscopically. 
b C578L6 mice were Infected In the tail vein with 1.5X10 4 live wild type B16 melanoma 
cells. 3 days later they were Infected subcutaneously with lxI0 b Irradiated wild type 
or Irradiated GM-CSF transduced HI 6 melanoma cells. 28 days later, lungs were 
removed and examined both microscopically and microscopically. 
The general conclusion from the results of therapeutic experiments in 
these murine models is that it is possible to cure animals burdened with a 
relatively small amount of pre-established tumor which could be equated with 
the micro-metastatic disease that is encountered frequently in the clinic. If 
so, it appears unlikely that radio-graphically detected metastatic disease in 
humans (tumor burden >10 9 cells) will be eradicated by the highest vaccine 
doses used in this trial. Therefore, if safety and immunogenic potency is 
established in this trial, studies of efficacy might best be pursued with 
adjuvant therapy for patients with minimal tumor burdens (e.g., following 
surgical resection) who are at high risk of recurrence because of the presence 
of micro-metastatic tumor. In the U.S., over one third of patients undergoing 
radical nephrectomy for RCC with curative intent relapse from micro-metastatic 
disease . 2 
2.3 FEASIBILITY OF PRODUCING A GM-CSF SECRETING RCC TUMOR VACCINE 
Feasibility of establishing and transducing primary human tumor 
cultures . 
The methods proposed are adequate to establish and transduce each 
patient's RCC cells following nephrectomy. Before submitting this protocol to 
the Johns Hopkins Joint Committee on Clinical Investigation, feasibility 
studies were conducted within the clinical research setting of the Johns 
Hopkins Oncology Center and Department of Urology where the trial is to take 
place. Of 29 consecutive RCC specimens received at the time of surgical 
excision, 25 were established as primary cell cultures. Among the 4 specimens 
not established were two benign tumors, one tumor contaminated at the time of 
resection, and 1 oncocytoma. Of the 25 tumors that were established as 
primary cultures, 22 of these patients had histologic subtypes that would have 
made them eligible for this trial. The 3 of 25 patients that would probably 
be ineligible for our trial had oncocytomas. Patients with oncocytomas will 
not be eligible for this study because these tumors have a benign course, and 
are rarely discovered as stage III or IV disease. Finally, of the 22 
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