3.4.b. 
GM-CSF Gene Expression in Vaccine Cells 
Prior to cell therapy, tumor vaccine cells prepared for patients 
assigned to the non-transduced arm of the study will be assayed for native GM- 
CSF secretion by ELISA and data recorded. In the GM-CSF gene transduced arm of 
the trial, secretion of GM-CSF will be determined by ELISA prior to first 
injection and set at 24-48 ng/10 6 vaccine cells/24 hr by the methods described 
in Appendix 14. C. 
3.4. c. Schedule of Vaccine Administration 
Cell therapy will be administered every 28 days for up to 3 
complete courses. Spacing and location of vaccination sites are detailed in 
Table 13. The first dose for each patient will be administered as an 
inpatient, followed by 24-hour observation before discharge. If no immediate 
toxicities are observed, the subsequent two vaccinations will be given in the 
Johns Hopkins Oncology Center Outpatient Department with 8-hours of clinical 
observation prior to release for scheduled follow-up visits. 
3.4. d. Vaccine Preparation and Formulation 
Vaccine preparation, formulation, and quality assurance protocols 
are delineated in Appendix 14. C. Vaccines will be finally formulated in the 
Johns Hopkins Oncology Center Pharmacy and placed in 10 ml graduated syringes. 
Vaccine will be administered using a 23 gauge needle as described in Table 13. 
TABLE 13 
INJECTION 
TECHNIQUE 
DOSE LEVEL 
DOSE INJECTED 
CELLS/ml 
INJECTION VOLUME 
INJECTIONS 
4 
1X10 9 
5X10 7 
0 . 5cc/site 
4 (2id/2sq) 
7.5X10 7 
1 . Occ/site 
12 (6id/6sq) 
3 
4X10 8 
5X10 7 
0 . 5cc/site 
16 (8id/8sq) 
2 
4xl0 7 
5xl0 7 
0 . 2cc/site 
4 (2id/2sq) 
1 
4X10 6 
5X10 6 
0 . 2cc/ site 
4 (2id/2sq) 
Injections with a 23 gauge needle will be given in the patients' 
forearms and thighs following a grid pattern prescribed by a customized foam 
rubber template. On the grid each injection is 3 cm at needle entry from 
nearest neighbor injection. 50% of total injections will be intradermal; 50% 
subcutaneous — alternating intradermal/subcutaneous in the cephalad-caudad 
axis. Cycle 1 vaccination limb will be right anterior thigh. Cycle 2 
vaccination limb will be non-dominant arm anterior fore-arm. Cycle 3 
vaccination limb will be left anterior thigh. 
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