4.0 CLINTCAT. PROTOCOL 
aRB# CCI# 93-103) 
A PHASE I STUDY OF IMMUNIZATION WITH GAMMA INTERFERON 
TRANSDUCED NEUROBLASTOMA CELLS 
1.0 GOALS AND OBJECTIVES 
Active specific immunotherapy with neuroblastoma cells that have been genetically engineered to express cytokines 
may evoke an effective anti-tumor immune response by improving the microenvironment in which immune system 
cells can recognize and respond to tumor associated antigens. Neuroblastoma cells frequently do not express class I 
or II MHC molecules and thus are likely to be poorly immunogenic. We will transfect amphotrophic retroviral 
vectors which express interferon-y (IFN-y) cDNA into autologous or partially HLA matched allogeneic 
neuroblastoma cells to render them producers. IFN-y has been shown to enhance expression of major 
histocompatibility complex (MHC) molecules on cell surfaces, which can result in their increased presentation of 
antigenic peptides to T lymphocytes. If IFN-y transduction increases expression of MHC, tumor associated pq)tides 
may be more effectively presented by neuroblastoma cells to the immune system. In addition, a microenvironment 
of IFN-y may facilitate processing and presentation of tumor associated antigens by macrophages to T lymphocytes. 
High risk neuroblastoma patients will be studied in this phase I/Ib toxicity/immunogenicity trial. They include 1) 
those who have undergone myeloablative therapy and autologous bone marrow transplantation (ABMT) who have no 
detectible disease but who are at significant risk for recurrence; and 2) those with p^sistent or recurrent disease. Our 
aims are as follows: 
1.1 To determine the maximum tolerable dose of IFN-y transduced autologous or allogeneic human 
neuroblastoma cells when given as a subcutaneous injection. 
1.2 To determine the local, regional, and systemic toxicides of EFN-y transduced neuroblastoma cells. 
1.3 To determine if IFN-y producing tumor cells elicit an and-neuroblastoma response in vivo as measured by 
standard clinical tests and immunocytologic evaluation of marrow metastases. 
2.0 BACKGROUND 
Neuroblastoma, which is a tumor of the peripheral nervous system, is the most common extracranial solid tumor of 
childhood and has an incid^ce of one per 7,(X)0 children und^ the age of 15 years.! Neuroblastomas differ markedly 
in their clinical behavior, with low, intermediate, and high risk groups of patients being defined at diagnosis based 
upon expected outcome following conventional therapy.^ Forty percent of children with neuroblastoma have low or 
intermediate risk tumors, and they nearly all become long-term, disease-free survivors following conventional 
therapy which includes surgery, lo^ irradiation, and chemotherapy. However, 60% are at high risk for developing 
fatal progressive disease 3-8. Our long-term goal is to improve the outcome of patients with high risk 
neuroblastoma. Although effective induction chemotherapy followed by high-dose, myeloablative 
chemoradiotherapy and ABMT achieves a complete remission in approximately 80% of high risk patients, 50% of 
these develop recurrent disease. Biologically based therapy, such as in this protocol, may be able to eliminate the 
relatively small number of tumor cells that survive maximtd chemotherapy, surgery, and irradiation. 
2.1 Preclinical Studies. Expression of MHC class-I molecules appears to be a critical determinant in rejection 
of tumor cells, since target cells must process and present peptide antigens in the context of class I molecules in 
order to be recognized and killed by CD8+ T lymphocytes.9 
IFN-y can increase the expression of MHC molecules in a variety of cell types. Antitumor effects have been 
observed after immunization of mice with IFN-y transduced fibrosarcoma cells. IFN-y secretion by CMS5 tumor 
cells decreased their tumorigenicity and induced antitumor immunity.!® This speared to be due to T cell immunity 
as judged by rejection of CMS 5 tumor challenge and persistence of specific cytotoxic activity in the spleens of 
inoculated mice. Expression of IFN-y cDNA in the Cl 300 mouse neuroblastoma cell line both enhanced MHC 
class-I expression and augmented antitumor immunity in the recipient!!*! 2 The nonimmunogenic, low MHC class 
I-expressing methylcholanthrene (MCA) induced sarcoma cell line, MCA 101, is a poor presenter of endogenously 
generated viral antigens to CD8+ T lymphocytes and cannot be used to generate tumor infiltrating lymphocytes. 
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