procedures. In order to handle large-scale tissue cultures, we plan to use 
automated systems for solution transfer and cell harvesting. A solution transfer 
pump, life cell tissue culture bags and the Celltrifuge II (Fenwall, Deerfield, IL) 
will be utilized. 
6.5.C Harvesting Conditions 
One out of ten bags will be tested for sterility 48 hrs. before 
harvesting by obtaining an aliquot of cells and sending it for culture by the 
Microbiology Laboratory of the University Hospital. Bacterial monitoring will 
include thioglycolate broth, chocolate agar and sheeps blood agar. Cell 
suspensions from the culture bags will be harvested and washed in phenol-free 
HBSS using the Celltrifuge II (Fenwal). A gram stain of the final cell pellet will 
be evaluated to confirm that there are no organisms prior to infusion. 
6.6 Adoptive Cellular Therapy and IL-2 Administration 
The lymphocytes will be suspended in infusion media consisting of 190 ml 
normal saline, 10 ml human albumin (concentration 25%, final concentration of 1.25%) and 
450,000 IU IL-2. IL-2 will be administered i.v. to enhance the activity of the activated cells 
and will be given 3 times daily for five days starting on the day of cell transfer. Infusions of IL- 
2 will be 360,000 lU/kg and will commence immediately following cell transfer. This schedule 
and dose of IL-2 has been associated with manageable and reversible side effects according 
to our previous experience with this regimen. The patients will be treated as inpatients in the 
Clinical Research Center. 
7.0 TOXICITIES TO BE MONITORED/DOSAGE MODIFICATIONS 
. 
7.1 Vaccination with retroviral transduced tumor cells 
Inoculation with a retrovirus-transduced tumor cell vaccine could theoretically pose 
risks to the patient which include: a. Possible exposure to replication competent retrovirus, 
b. Possible adverse effects from provirus integration, c. Growth of the inoculated tumor at 
tumor vaccine site, d. Systemic toxicity from IL-4 production by the tumor cell vaccine, and e. 
Generation of autoimmunity by the tumor cell vaccine. 
7.1. a Possible exposure to replication competent retrovirus . 
Retrovirus transduction of tumors for vaccine preparation will only be performed with 
FDA approved retrovirus stocks in a dedicated facility (the Human Applications Lab) located 
in the Clinical Research Center at The University of Michigan. All retrovirus stocks will be 
safety tested and documented to be free of replication competent virus, bacterial, viral, and 
fungal adventitial agents before use in tumor transduction for clinical trials. 
7.1. b Possible adverse effects from provirus insertion . 
The tumor cells will be irradiated prior to administration to patients. This should 
provide for the complete loss of ability of the tumor cells to divide and or persist in the 
patients. Thus no growth of the tumor cells is expected, and therefore any possible 
mutagenesis from provirus integration should not result in harmful effects. If any tumors do 
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