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Flow cytometry for clinical estimation of circulating hematopoietic
progenitors for autologous transplantation in cancer patients
S Siena, M Bregni, B Brando, N Belli, F Ravagnani, L Gandola, AC Stern, PM Lansdorp, G Bonadonna and AM Gianni
Cristina Gandini Transplantation Unit, Istituto Nazionale Tumori, Milan,
Italy.
Optimum methods of harvesting circulating hematopoietic progenitors for
autologous transplantation to support myeloablative cancer therapy are
still uncertain, mostly because of the lack of an assay for marrow-
repopulating stem cells. The CFU-GM assay, the commonly used indirect
indicator of the quality of the graft, is poorly standardized and provides
results evaluable only retrospectively. Based on the knowledge that
hematopoietic progenitors express CD34 and CD33 differentiation antigens,
we developed a dual-color direct immunofluorescence flow cytometry assay
with the aim of replacing the CFU-GM assay advantageously. For this
purpose, we applied both assays to 157 blood samples obtained daily
throughout 20 different recoveries from pancytopenia induced by high-dose
cyclophosphamide or etoposide cancer therapy with or without recombinant
human GM colony-stimulating factor (rhGM-CSF). The appearance of CD34+
cells in the circulation indicated that hematopoietic progenitors had
increased to more than 500 CFU- GM/mL, a level clinically adequate for
large-scale harvest by leukapheresis. Total CD34+ cells correlated well
with CFU-GM (r = .89), and data could be fitted by a linear regression line
described by the equation y = 388.3 + 64.0x, where y = CFU-GM/mL and x =
CD34+ cells per microliter. Moreover, in a series of six patients treated
with myeloablative chemoradiotherapy, early hematopoietic recovery of
marrow functions was predicted more accurately by the number of
transplanted CD34+/CD33+ cells than by either total nucleated cells,
CFU-GM, CD34+/CD33- cells, or CD34-/CD33+ cells. Data presented in this
article favor clinical use of the CD34/CD33 flow cytometry assay to guide
harvesting of circulating hematopoietic progenitors for autologous
transplantation and contribute to better understanding of the role played
by circulating hematopoietic progenitor cell subsets in marrow recovery
after myeloablative cancer therapy.
Volume 77,
Issue 2,
pp. 400-409,
01/15/1991
Copyright © 1991 by The American Society of Hematology
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