function after intensive VP16, cytoxan, and total body 
irradiation (DM90-064) . Multiple bone marrow aspirations 
from the iliac crests will be performed before cytoxan, 
VP-16 and TBI to store twice the engrafting dose of bone 
marrow (2 x 10 8 /kg nucleated cells) . A second bone 
marrow storage will be considered if less than 2 x 10 8 
total nucleated cells/kg were collected in the first 
storage unless a back-up marrow is available. Another 
criterion for adequacy of the marrow collection will be 
a total of 2 x 10 4 CFUGM/kg. 4 x 10 8 /kg nucleated cells 
and 2 x 10 4 CFUGM/kg from the peripheral blood will also 
be collected for reconstitution. Peripheral blood stem 
cells will be collected by continuous flow centrifuge 
during hematopoietic recovery (wbc 0. 3-1. 0xl0 3 /mmr) 
following daunomycin, and high dose ara-c or other 
equivalent chemotherapy. 
The peripheral blood and marrow will be subjected to CD34 
positive selection. A total of 0.7 x 10 6 CD34 positive 
cells/kg must be available. The retroviral marking will 
be performed as described in Appendix G and as follows: 
Thirty percent of the nucleated peripheral blood or 
marrow cells remaining after purging will be incubated 
with the LNL-6 or GIN vectors respectively at the time of 
storage before cryopreservation. For every nucleated 
peripheral blood or marrow cell obtained after Ficoll 
Hypaque separation, 10 vector particles (multiplicity of 
infection units) produced by Genetics Therapy, Inc. of 
Gaithersburg, Maryland, as an FDA approved product for 
this purpose, will be added with the nucleated cells and 
incubated for six hours (see Appendix G) . The final 
concentration of the cells in the vector incubation will 
be lxlO 6 cells/cc. The choice of the dose of the LNL-6 
vector or GIN vector (10 viral infectious units/cell) was 
based on previous experience by Dr. F. Anderson and Dr. 
M. Brenner with this virus. The cells will be rinsed and 
a small portion (10 6 ) cultured in a CFUGM assay and the 
majority of cells will be frozen and reinfused with the 
rest of the autologous marrow and peripheral blood cells. 
5.3 Treatment Plan: All patients should be treated in the 
Protected Environment if available. 
a. The preparative ablative regimen will consist 
of the following systemic chemotherapy: 
Cyclophosphamide (see Appendix D) : 60 mg/kg in 0.5 
liter D5W intravenously over 3 hours daily for 2 
days - days 1 to 2 (total 120 mg/kg) , mesna, 10 
mg/kg will be given q4h during cyclophosphamide 
infusion and for 24 hours following the end of the 
mesna infusion. 
Recombinant DNA Research, Volume 15 
