Stable Disease (SD): Less than a 50% decrease in the product of the 
perpendicular diameters of the tumor relative to the baseline evaluation 
without the appearance of any 'new areas of disease. 
Progressive Disease (PD): Greater than a 25% increase in the products of the 
perpendicular diameters of the tumor relative to the baseline evaluation, or 
the appearance of any new areas of disease. 
6.2 Relapse 
If relapse occurs then NB cells will be separated from the marrow by FACS 
analysis separating cells which are CD34', CD45' and UJISA"^. It will be 
determined by PCR whether or not NB cells contain the marker genes. We 
will also determine whether the relapse has occurred from a single clonogenic 
progenitor or whether many cells have the ability to re-populate the patient. 
This question will be answered by analyzing the site where the marker gene 
has inserted using Southern blotting and RFLP and inverted PCR (see 
Technical Appendix). 
6.3 Regeneration of Cryopreserved Autologous Marrow on Individual Colonies 
Patients will be monitored at weekly intervals for 6 weeks, monthly intervals 
for 6 months and annually thereafter for 5 years for the presence of cells 
containing the marker genes in the peripheral blood (PB) using FACS sorted 
populations. PB will be separated into T cells, B cells, monocytes and 
granulocytes using appropriate MAb, and each lineage examined for gene 
expression and for marker gene insertion sites by PCR and Southern blotting. 
To exclude the possibility of false positive PCR by low level contamination of 
one lineage by another, the amplification will be undertaken as quantitatively 
as possible. 
When marrow is obtained to assess engraftment and disease 
response/progression, additional cultures will analyze whether marker genes 
are present in CD34+ progenitor cells and of what lineage. 
Changes in marker gene expression in cells of different lineages will be 
correlated with changes in endogenous (serum) cytokine levels including TFN, 
ylFN, IL3, and GM-CSF (measured by commercially available immunoassays), 
induced by (e.g) infection or chemotherapy. Ultimately changes will be 
correlated with the effects of exogenous recombinant cytokines such as IL2 
and IL3, which will be components of future submitted protocols used to 
treat these diseases. 
Recombinant DNA Research, Volume 14 
[719] 
