C. WHAT QUESTIONS ARE ANSWERABLE BY THIS PROTOCOL: 
i) Does relapse of accelerated phase or blastic crisis come 
from leukemia cells remaining as residual systemic disease in the 
patient following TBI, VP-16, and cyclophosphamide therapy, or does 
it arise from residual blastic leukemia cells remaining in the 
transfused autologous marrow? 
ii) When relapse from the infused autologous stem cells 
occurs, is it clonal or nonclonal? The latter question can be 
answered by studying if there is more than one type of marker 
integration site in the blastic or accelerated phase leukemic cells 
which appear at the time of relapse. The clonal ity of normal 
hematopoietic reconstitution following transplantation can also be 
studied by this method. 
iii) Since some patients will be in complete cytogenetic 
remission at the time of autologous stem cell collection, the 
question as to the origin of cytogenetic relapse (marrow/peripheral 
blood infusion versus sytemic disease) can be answered. 
B. RESEARCH DESIGN. ANTICIPATED RISKS. AND BENEFITS : 
1. STRUCTURE AND CHARACTERISTICS OF THE BIOLOGICAL SYSTEM: 
a. WHAT IS THE STRUCTURE OF THE CLONED DNA THAT WILL BE USED? 
We have chosen to use the LNL6 vector previously evaluated and 
approved by the NIH RAC committee and the FDA for marking purposes. 
The LNL6 vector has been used in the N2 TIL protocol of Rosenberg 
and Anderson (1) which was originally developed in the laboratory 
of A.D. Miller (2) . This vector is obtained as an FDA-approved 
supernatant developed from the PA317 producer cell line by Genetic 
Therapy, Inc. of Gaithersburg, Maryland. This vector is also 
approved for use in a similar form at St. Jude's Hospital for AML 
led by Dr. Malcolm Brenner. 
b. WHAT IS THE STRUCTURE OF THE MATERIAL THAT WILL BE 
ADMINISTERED TO THE PATIENT? 
bl . Describe the preparation, structure, and composition of 
the materials that will be given to the patient or used to treat 
the patient's cells : 
A cell-free supernatant from the PA317 producer cell lines 
which contains 10^-10^ MOl of the LNL6 vector, was obtained from 
Genetic Therapy, Inc. This preparation is approved by the FDA for 
use in treating human lymphocytes and marrow to be infused into 
patients. The viral supernatant is suspended in DMEM tissue culture 
medium. The supernatant will be incubated with the patient's marrow 
cells. This preparation of LNL6 is identical to that previously 
approved by the RAC Committee and utilized by Stephen Rosenberg and 
W. French Anderson in the N2-TIL gene transfer clinical protocol 
(1) . It has also been approved for use in AML autologous marrow 
setting by Malcolm Brenner of St. Jude's Hospital. The DNA 
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Recombinant DNA Research, Volume 14 
