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Circulation of CD34+ hematopoietic stem cells in the peripheral blood of
high-dose cyclophosphamide-treated patients: enhancement by intravenous
recombinant human granulocyte-macrophage colony-stimulating factor
S Siena, M Bregni, B Brando, F Ravagnani, G Bonadonna and AM Gianni
Cristina Gandini Transplantation Unit, Istituto Nazionale Tumori, Milan,
Italy.
We report that hematopoietic progenitor cells expressing the CD34 antigen
(CD34+ cells) transiently circulate in the peripheral blood (PB) of cancer
patients treated with 7 g/m2 cyclophosphamide (HD-CTX) with or without
recombinant human granulocyte macrophage-colony stimulating factor
(rHuGM-CSF). In adult humans, CD34+ cells represent a minor fraction (1% to
4%) of bone marrow (BM) cells, comprising virtually all hematopoietic
colony-forming progenitors in vitro and probably also stem cells capable of
restoring hematopoiesis of lethally irradiated hosts. We show that CD34+
cell circulation is fivefold enhanced by rHuGM-CSF 5.5 protein
micrograms/kg/day by continuous intravenous infusion for 14 days after
HD-CTX. During the third week after HD-CTX (ie, when CD34+ cells peak in
the circulation), large- scale collection of PB leukocytes by three to four
continuous-flow leukaphereses allows the yield of 2.19 to 2.73 x 10(9) or
0.45 to 0.56 x 10(9) CD34+ cells depending on whether or not patients
receive rHuGM- CSF. The number of CD34+ cells retrieved from the
circulation by leukaphereses exceeds the number that can be harvested by
multiple BM aspirations under general anesthesia. Thus, after therapy with
HD-CTX and rHuGM-CSF, PB represents a rich source of hematopoietic
progenitors possibly usable for restoring hematopoiesis after myeloablative
chemoradiotherapy. To determine whether CD34+ cells found in the PB are
equivalent to their marrow counterpart, we evaluated their in vitro growth
characteristics and immunological phenotype by colony assays and dual-color
immunofluorescence, respectively. We show that PB CD34+ cells possess
qualitatively normal hematopoietic colony growth and high cloning
efficiency comparable to that observed with BM CD34+ cells. In addition, PB
CD34+ cells display heterogeneous surface membrane differentiation antigens
analogous to BM CD34+ cells. The availability of large quantities of CD34+
cells by leukapheresis is relevant to the field of stem cell
transplantation and possibly to genetic manipulations of the hematopoietic
system in humans.
Volume 74,
Issue 6,
pp. 1905-1914,
11/01/1989
Copyright © 1989 by The American Society of Hematology
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