antigen in an appropriate MHC class-I context This could lead to much more simplified tumor vaccination schemes 
and potentially alleviate the need for cytokine introduction into individual autologous tumor cells. 
The retroviral vector used in these studies is based upon Moloney murine leukemia virus, which has been engineered 
to replace its extraneous viral coding sequences with the human EFN-y cDNA. Unlike true viruses, the recombinant 
vector is incapable of replication since it lacks all the viral structural genes. A detailed characterization of the vector 
gene construct will be presented to the RAC and the FDA for approval prior to initiation of the study. 
6.12 Formulation and Storage. IFN-y transduced tumor cells are supplied frozen viably in L15 tissue 
culture medium with 2.5% human serum albumin, 3% Hespan, and 20% dimethylsulfoxide at concentrations 
appropriate to provide viable thawed cells for the dose levels indicated. For example, freezing 1.5 x 10^ cells in 1 
ml of medium would yield approximately 10^ cells on thawing (anticipated recovery of 66%); this would provide 
sufficient cells for one course for a 15 kg patient (approximately 0.66 m2) receiving the level 1 dose. Viably frozen 
cells will be stored in sealed, sterile tubes in liquid nitrogen vapor and only removed immediately befwe injection. 
6.13. Stability. Once viably frozen, cells are stable indefinitely, provided they are not removed from liquid 
nitrogen vapor. 
6.14 Preparation for Injection. Cells will be rapidly thawed by partial immersion of the vial in a 37 
<>C water bath. Once thawed, they will be washed once, assessed for viability, and resuspended at an appropriate 
concentration in 1 ml of LI 5 medium containing 2.5% human serum albumin. Without delay, thawed cells will be 
irradiated (5000 cGy) and injected into the patient 
6.15. Administration. The appropriate total cell dose will be divided into two aliquots and injected into 
two sites (alternate thighs with arms) subcutaneously. The volume of cells injected generally will be 0.25 - 0.5 ml 
in each site. The cell concentration will be <50 x lO^ per ml to prevent excess cell aggregation, and so it may be 
necessary to inject more than 2 sites for some patients. 
7.0 CRITERIA FOR REMOVAL FROM PROTOCOL THERAPY 
7 . 1 Any patient who develops progressive disease. 
7 . 2 Any patient who develops unmanageable local or regional toxicity. 
7 . 3 Any patient who develops grade 3 (X 4 toxicity that does not resolve within 28 days. 
7 . 4 Completion of 7 courses of therapy (weeks 0-36). 
7 . 3 Allergic reaction to IFN-y or life-threatening allergic reaction to IFN-y transduced tumor cells. 
7.4 At discretion of patient or physician. 
8.0 SUPPORTIVE CARE 
Supportive care will be given according to the Judgement of the treating physician. 
9.0 IMMUNOBIOLOGIC AND MARROW STUDIES 
9 . 1 Patient Considerations. In addition to routine tests for evaluating toxicity, the following selected 
tests will be performed. These will provide information about the patient's immune status, immunologic response 
to the tumor cell injection, interferon level, and possible anti-tumor effect 
9.2 Sampling Schedule. The following studies will be done before and after immunization on weeks 
0, 8, 24, and 36: 
9.21 Blood. Absolute numbers of T cells, T cell subsets, and B cells; response to PHA. 
9.22 Blood.: T cell and CD8+ cytotoxicity (direct and after in vitro expansion with cytokine producing, 
irradiated immunizing turntx cells) for autologous and allogeneic neuroblastoma cells. 
9.23 Bone marrow, quantitative immunocytologic analysis for neuroblastoma cells involvement (only for 
patients with detectible metastases at study entry). Bone marrow examination should be bilateral aspirates, no 
biopsies needed. 
Recombinant DNA Research, Volume 19 
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