line. 28 ’ 38 ’ 53 ’ 15 ' 17 ’ 24,31,34,35 Thus, neuroblastoma cells are sensitive to autologous 
IL-2 induced activated killer cells in vitro, and our own data show that killing 
can be induced even when effector cells are obtained soon after ABMT. The 
process is mediated independently of MHC recognition or of CD3 receptor 
involvement, since MAb to these structures do not affect the cytotoxic 
response. Secondly, Phase I studies of IL-2 or IL-2-induced TIL cells have 
been undertaken in children with advanced neuroblastoma, including patients 
relapsing after ABMT. One complete and four partial remissions were 
observed in 29 patients studied. Finally, IL-2 induces gamma interferon 
release from CD16+/- cells in vitro and in vivo. 46 This secondary production 
of IFN-gamma may subsequently upregulate Class I HLA expression on 
neuroblasts, increase immunogenicity and thereby further augment the effects 
of IL-2. 26 
4.0 TRANSDUCTION OF NEUROBLASTOMA CELLS 
Tumor cells from all patients with Stage C and D disease will be established 
as cell lines at the time of presentation. Neuroblastoma cell lines are grown 
out as described in Appendix A and are transduced with the GlNCVi2.23 
vector, a modified retrovirus closely related to the LNL6 backbone used in the 
previous St. Jude gene marking protocols, NEBREL and NEBREM. This 
vector contains the same neomycin resistance gene, but in addition the 
multicloning site now contains sequences encoding the human interleukin-2 
gene and a CMV promotor. 
LNL6 has been extensively studied in rodent models and in non-human- 
primates and was approved for use in man in 1989. The results of the first in 
vivo human study on transduced cells have been reported, and our own safety 
data using marrow cells as the vector target are being prepared for publication. 
The safety data for LNL-6 and for the vector to be used in the current study 
are contained within the Drug Master Files # BB-MF-3886 and BB-MF-4195 
which were used for granting our institutional IND’s 3838 (LNL6) and 4272 
(GIN). A new IND will be requested for the IL-2 modified GIN vector, and 
the current proposal will cross reference these data. The major safety concerns 
and their resolution have been dealt with at length in previous protocols. 48 ' 50 
5.0 PATIENT ELIGIBILITY (See Section 1.0) 
5.1 All patients under 21 years of age at diagnosis with neuroblastoma 
unresponsive to conventional therapy shall be eligible for this protocol 
Recombinant DNA Research, Volume 15 
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