regimens used. One of the most important potential advantages of the double gene 
marking approach is that each patient acts directly as their own control for the effects of 
growth factors, greatly increasing the power of the analysis on even a limited number of 
patients. This issue is addressed in the Statistical Section. Because each patient will act 
as their own control differences in underlying diseases and the degree of ablation the 
conditioning regimen will induce are not relevant. 
4.0 STUDY OUTLINE 
Marrow will be harvested as patients recover from previous chemotherapy. At least 3 x 
10* cells/kg will be harvested. 10* cell/kg will be frozen as a safety backup and the 
remaining cells will be processed on the CellPro column to purify CD34 cells. The 
purified CD34 population will be split into two aliquots which will randomly be marked 
with the LNL6 or GlNa vector. One aliquot will be frozen without further manipulation 
while the remaining fraction will be incubated in the presence of growth factors for 5-7 
days and then frozen. Patients will be conditioned with the regimen specified on their 
disease based protocol and both aliquots of CD34 cells will then be administered 
simultaneously at the time of transplant. We will use semiquantitative PCR to detect the 
presence of both marker genes in patients blood and marrow at various intervals post 
transplant. 
Outline of Protocol 
1/3 marrow 
Safety backup 
2/3 marrow 
CD34 selection 
Mark with LNL6 
Mark with GlNa 
Cryopreserve 
or 
Expand ex vivo with 
growth factors 
Expand ex vivo 
with growth factors 
or 
Cryopreserve 
Reinfuse both aliquots at time 
of BMT 
Recombinant DNA Research, Volume 19 
[493] 
