will be held constant unless dose modification is needed for toxicity (Section 7.0). A 
CBC with differential will be obtained two times per week while receiving taxol or 
vinblastine therapy. 
5. 1 1 Patients will be treated with taxol or vinblastine and G-CSF until stable disease has 
been documented over 3 cycles or until progressive disease. 
6.0 Cell Harvesting, Processing, Transduction, and Reinfusion 
6. 1 Peripheral Blood Stem Cell Collection 
6.1.1 Patients will undergo daily apheresis on a continuous flow apheresis 
instrument such as a Spectra 2991 (COBE) or a Fenwal CS3000 (Baxter) 
following cytoxan/G-CSF administration beginning on the day when the total 
leukocyte count reaches 1000/mm 3 . The first day's collection will be processed 
and cryopreserved as per the original protocols. The collection on the day after 
the collection is first > 1 X 10 8 /kg nucleated cells/kg will be CD34-enriched 
and transduced with the MDR-1 vector. No further processing of the collected 
cells is performed prior to the CD34-enrichment procedure (Section 6.3). 
6.1.2 Collections on subsequent days will proceed as necessary until > 3 X 10 8 total 
nucleated cells/kg are collected. These collections will be processed and 
cryopreserved directly without CD 34-enrichment or transduction. 
6.1.3 If no collection is > 1 X10 8 nucleated cells/kg, then the patient will not have 
peripheral blood cells CD34-enriched or genetically modified. 
6.2 Bone Marrow Harvest 
6.2. 1 The following preoperative tests must be obtained prior to harvest: CBC, 
platelets, PT, PTT, electrolytes, BUN, Cr, glucose, calcium, phosphate, 
albumin, magnesium, liver function tests, urinalysis, Chest X-ray, ECG. 
Type and Cross 3 units irradiated PRBC for the OR. 
6.2.2 Patients' bone marrow will be harvested after 4-5 cycles of induction 
chemotherapy and after the PBSC harvest has been done as below. Bone 
marrow harvest is arranged by contacting Dr. Wyndham Wilson (496-6404 or 
page 104-2399-7) or George Bryant (496-9521 or page 104-2556-7) at least 1 
week before the procedure. Patients must have an AGO 1500/mm 3 and 
platelets >100,000/mm 3 . Marrow collection is performed in the Clinical 
Center operating rooms under general anesthesia. The patient must be admitted 
to the hospital by 2:00 P.M. on the day prior to marrow collection and 
evaluated by the anesthesiologist and BMT team. 
6.2.3 The entire marrow is processed on a cell processing instrument such as the 
Fenwal CS3000 or the Spectra 2991 to obtain a buffy coat preparation. ABMT 
requires a minimum of 5 x 10 7 mononuclear cells/kg body weight. In general, 
>lxl0 8 nucleated cells/kg body weight are harvested. Every attempt will be 
made to harvest >3x108 nucleated cells/Kg body weight before processing. If 
it is possible to obtain >1x108 nucleated cells/Kg before albumin processing, 
70% of the purified marrow will be immediately cryopreserved using standard 
techniques. The remaining 30% will be used for retroviral transfection. If <1 
x 10 8 nucleated cells/Kg body weight are harvested, the entire purified marrow 
will be frozen immediately. In this case, no bone marrow cells will be used for 
retroviral transduction. 
6.3 Antibody Treatment and Wash 
6.3.1 The primary buffy coat bone marrow or apheresis product is transferred to a 
600 ml bag for antibody incubation. One 3.0 mg vial of CellPro 12.8 
biotinylated murine IgM anti-human CD34 antibody is thawed at room 
temperature for 20 minutes. 
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Recombinant DNA Research, Volume 18 
