GTI0101 (04/08/93) 
Page 25 of 35 
5.33 Transduction and Transplantation Procedures : 
The retroviral vector supernatant produced in this study will be produced by 
Genetic Therapy, Inc. (GTI). GTI has produced the retroviral vector supernatant 
utilized in most of the gene transfer/ therapy studies performed. The vector 
supernatant used in this protocol will be produced and quality control tested the 
same as has been done for other retroviral vector supernatants. 
5.331 Transduction Procedure 
Study Days M1-M3. P5-P10 : After removal of aliqouts of enriched cells 
for studies, the remainder of the CD34-enriched population of cells will 
be transduced with the retroviral vector. Both the marrow-derived and 
peripheral blood-derived cells will be transduced under the same 
conditions. The separated CD34-enriched cells will be cultured (IX 
lOVml) in vector supernatant with IL-3, IL-6, and SCF and 4 /xg/ml 
protamine sulfate for a period of 72 hours. Every 24 hours the cells will 
be spun down and resuspended in fresh supernatant, growth factors, and 
protamine. After the period in culture, the cells will be washed and 
samples taken for analysis (gene transfer efficiency, GC enzyme activity, 
colony-forming ability, and replication competent retroviral assays). The 
remaining cells will be resuspended in heparinized medium for reinfusion. 
Included in Appendix VII is a sample certificate of the tests that will be 
performed on each hematopoietic stem cell sample prepared for reinfusion 
into the patient. 
5.332 Reinfusion of Transduced Cells 
Study Davs M4. P8-P10 : The transduced cells will be reinfused into the 
patient over a period of two hours or less. Premedication, given 30 min 
before infusion begins, will consist of 25 mg diphenylhydramine IV and 
650 mg acetaminophen given po (100 mg hydrocortisone IV is optional). 
Patients will be closely monitored during this process and several hours 
after. 
5.333 Post-Infusion Care 
Patients will be monitored in the hospital overnight. If no adverse 
reactions are observed, the patients will be discharged the next day (Study 
Days M5, PI 1) for subsequent out-patient follow-up. Further patients 
may receive the infusion as an out-patient. 
5.334 Frequency of Infusion of Transduced Cells 
The first infusion only may be marrow-harvested cells (if the bone 
marrow harvest method is chosen, the patients will undergo 1 treatment 
only). However, if the PBMC-harvest method is chosen, patients may 
receive up to four cycles (or receive PBMC-transduced cells four times). 
The cycles will be no more frequent than every 2 months. 
Recombinant DNA Research, Volume 17 
