autologous blood and marrow transplantation - Blog Science ...

438 - Chapter 7: Solid Tumors
Table 1. Clinical characteristics of the patients at the time of the study
Interval without
Age Previous chemotherapy Previous
Patient (years) Sex Diagnosis chemotherapy (months) radiotherapy
1 40 F BC — NA —
2 33 M Sarcoma — NA —
3 41 F BC CMFEV X 4, PLF X 3 21 —
4 40 F BC CMF X 13, PLF X 4 11 Yes
5 54 F BC CMF X 12 5 Yes
6 37 F BC GIP X 4 0.5 —
7 47 F Sarcoma — NA —
8 50 F BC GIP X 6 1 —
9 49 F BC CMF X 10 1 —
10 53 F BC CMF X 6, 1 —
TAX + VNRB X 9
BC, breast cancer; CMF, cyclophosphamide + methotrexate + 5-fluorouracil; CMFEV,
cyclophosphamide + methotrexate + 5-fluorouracil + epirubicin + vincristine; GIP, gemc-
itabine + ifosfamide + cisplatin; NA, not applicable; PLF, cisplatin + 5-fluorouracil +
folinic acid; TAX, taxol; VNRB, vinorelbine.
a daily basis. In all patients, at both cycles, complete blood counts, number of
CD34 +
cells, and progenitor cell assays were performed daily on days 0 to 10.
Flow cytometry
Ethylenediaminetetraacetic acid (EDTA)-anticoagulated blood was incubated
for 30 minutes on ice with 10 pL fluorescein isothiocyanate (FITC)- or
phycoerythrin (PE)-conjugated monoclonal antibodies. The following antibody
combinations were used: CD4-FITC/CD8-PE, CD45-FITC/CD14-PE, CD34-PE.
Each fluorescence analysis included a double negative isotype control (IgGj-
FITC/IgGj-PE). Antibodies and isotype controls were purchased from Becton
Dickinson (San Jose, CA). After red blood cell lysis with FACS Lysing Solution
(Becton Dickinson), samples were washed in Dulbecco's phosphate buffered saline
without Ca 2+
2 +
and Mg and acquired in listmode. Threshold was set on the forward
scatter (FSC) to exclude cellular debris. A minimum of 40,000 events and a
minimum of 100 CD34 +
events were acquired. Analysis was performed by setting
two gates. The first gate was set on a plot of CD45 vs. CD 14 and drawn to include
lymphocyte and monocyte cells. The second gate was set on the plot of FSC vs.
side scatter (SSC) to include lymphocyte and monocyte cells and was activated to
generate a plot of SSC vs. anti-CD34 fluorescence intensity. The percentage of
positive cells was determined by subtracting the percentage of fluorescent cells in