1 . 0 
OBJECTIVES 
1.1 To evaluate two separate purging steps in the marrow 
of CLL patients. 
1.2 To evaluate the origin of relapse of leukemia cells 
remaining in the systemic circulation following 
intensive preparative therapy versus leukemic cells 
remaining in the marrow following purging with 
monoclonal antibodies. 
1.3 The marrow will have been stored at a time of 
hematological remission following conventional dose 
chemotherapy . 
1.4 To analyze the hematological response following 
autologous transplant. 
2 . 0 BACKGROUND 
The use of exogenous hematopoietic reconstitution has 
permitted the delivery of intensive therapy for induction 
of remission and cures in both acute and chronic 
leukemias. Autologous and allogeneic bone marrow 
transplantation is designed to generate induction of long 
term disease-free remissions. 
Chronic lymphocytic leukemia is a disease in which the 
majority of patients relapse following induction of 
complete remissions induced by fludarabine and prednisone 
or fludarabine alone. For patients ineligible for 
allografting, the only option with a hope of long term 
disease-free survival is intensive therapy supported by 
autologous bone marrow transplantation. 
Marrow contamination by neoplastic lymphoid cells is a 
major problem even in patients in whom morphological 
remission is achieved, since virtually all of these 
patients exhibit marrow relapse. Thus, in autologous 
bone marrow transplantation, it is important to 
fractionate in vitro the bone marrow following 
achievement of a complete remission so as to remove the 
majority of neoplastic lymphoid cells. We are proposing 
to first induce morphological remission with the 
fludarabine and prednisone. Following harvesting of an 
engrafting dose of total nucleated cells, we will use 
CD34 selection to harvest an engrafting dose of early 
progenitor cells. Most of the normal differentiated 
lymphoid cells lack the CD34 marker. We will also use 
another in vitro monoclonal antibody fractionation step 
( CD19 negative selection) to reduce the ratio of CLL 
cells to normal cells. We will use in vitro monoclonal 
anibody bead conjugates to remove CD19 positive lymphoid 
. cells. 
Recombinant DNA Research, Volume 16 
[389] 
