grow at the inoculation site, they will be biopsied, and assayed for IL-4 production, and for 
abundance of provirus sequences. 
7.1 .c Growth of the inoculated tumor at tumor vaccine site. 
These patients will receive up to 10 8 /irradiated transduced tumor cells intradermally in 
one or two sites. Tumor cells will be irradiated with 5,000 cGy. In our previous experience, 
inoculations of 2 x 10 7 irradiated (2,500 cGy) tumor cells admixed with BCG has not resulted 
in tumor outgrowth in more than 20 patients. Berd et al. has also reported an extensive 
experience in 64 melanoma patients treated with multiple injections of 1-2.5 x 10 7 irradiated 
(2,500 cGy) autologous tumor cells plus BCG without evidence of tumor outgrowth at the 
injection sites (17). The vaccine sites will be carefully monitored, and if tumor growth does 
occur the sites have been selected so that they can easily be excised with minimal morbidity. 
A small chance does exist, however, that if this site does grow that it might lead to the spread 
of injected tumor cells to other sites of the body. This unlikely event should not negatively 
influence the prognosis of this patient population which already has documented metastatic 
disease with limited life expectancy. 
7.1. d Systemic toxicity from IL-4 production bv the tumor cell vaccine. 
If gene modified tumor cells grow and produce IL-4, then patients may be exposed to 
systemic IL-4. In theory, if a palpable tumor nodule were evident at 1 cm in size, 
approximately 10 9 cells would be present and secreting at least 50 ng of IL-4/24 hrs 
(minimum required level of IL-4 secretion is 50 pg/10 6 cells/24 hr). The maximum tolerable 
dose of intravenous bolus IL-4 per day has been reported to be 45 mcg/kg/24 hr (30). In a 50 
kg patient, the amount of IL-4 potentially secreted by 10 9 cells would represent less than 
0.01% of the maximum tolerable dose. We will monitor for any IL-4 toxicity by measurement 
of serum IL-4 levels. 
7.1. e Generation of autoimmunity by the tumor cell vaccine . 
If IL-4 production within an autologous tumor unlocks immunity to several self 
antigens, it may predispose the patient to develop autoimmunity and/or an autoimmune like 
syndrome. Tumor vaccines have been used throughout this century without such an effect, 
and in our BCG/autologous cell trial we have not seen evidence of clinical autoimmunity. We 
will measure anti-DNA antibodies to monitor for this possibility. 
7.2 Cellular Therapv/IL-2 Administration 
In all clinical studies of adoptive immunotherapy the major toxicities have been 
generally attributable to the infusion of high-dose IL-2 (ie. 720,000 lU/kg i.v. q8h). At this 
dose, the most prominent side effects are hypotension, leaky capillary syndrome, fluid 
retention, oliguria, respiratory distress and mental confusion (1 ,2). Since the side effects are 
dose related, it is anticipated that the toxicity associated with this proposal should be less 
since the dose of IL-2 being administered is 360,000 lU/kg i.v. q8h for a total of 5 days. With 
our current clinical studies, we have treated 14 patients with activated vaccine-primed LN 
cells and IL-2 at the proposed dosage without life-threatening toxicities. 
Medications are used to overcome some of the side effects of treatment. Fever, chills 
and malaise are significantly reduced by the use of acetaminophen (650 mg every six hours) 
and indomethacin (25 mg every six hours). Many patients receive hydroxyzine 
hydrochloride, an antihistamine, for treatment of a generalized erythematous rash. Patients 
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