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Collection, Tumor Contamination, and Engraftment Kinetics of Highly
Purified Hematopoietic Progenitor Cells to Support High Dose
Therapy in Multiple Myeloma
G. Tricot,
Y. Gazitt,
T. Leemhuis,
S. Jagannath,
K.R. Desikan,
D. Siegel,
A. Fassas,
S. Tindle,
J. Nelson,
C. Juttner,
A. Tsukamoto,
J. Hallagan,
K. Atkinson,
C. Reading,
R. Hoffman, and
B. Barlogie
From the Division of Hematology/Oncology and the Arkansas Cancer
Research Center, University of Arkansas for Medical Sciences, Little
Rock, AR; and Systemix Inc, Palo Alto, CA.
Unfractionated peripheral blood stem cell (PBSC) grafts contain
measurable quantities of myeloma cells and are therefore a potential
source of relapse posttransplantation. In contrast, fluorescence-activated cell sorting (FACS)-sorted CD34+
Thy1+ Lin peripheral blood
cells are substantially enriched for stem cell activity, yet contain
virtually no clonal myeloma cells. A study was performed in patients
with symptomatic myeloma, who had received 12 months or less of
preceding standard chemotherapy, to evaluate the feasibility of large
scale purification of primitive hematopoietic stem cells in order to
study engraftment kinetics posttransplantation and the degree of tumor
cell contamination of this cell population, based on polymerase chain
reaction (PCR) analysis for the patient-specific complementarity-determining region III (CDR III). PBSC were mobilized with high dose cyclophosphamide and granulocyte-macrophage
colony-stimulating factor (GM-CSF). A combination of elutriation and
chemical lysis was used to deplete PBSC collections of monocytes,
granulocytes, erythrocytes, and platelets. Subsequently,
CD34+ Thy1+ Lin
progenitor cells were purified with high speed cell sorting. Of the 10 evaluable patients, nine met the required minimum criteria of  7.2 × 105 cells/kg to support tandem transplants. After high
dose melphalan (200 mg/m2) eight engrafted successfully,
although granulocyte (absolute neutrophil count [ANC]
>0.5 × 109/L, 16 days) and platelet recovery
(platelets > 50 × 109/L, 39 days) was substantially
delayed when compared with unmanipulated PBSC grafts; one patient
required infusion of a reserve graft because of lack of evidence of
engraftment by day +28. Three patients proceeded to a second graft
with high dose melphalan and total body irradiation; two required
infusion of a reserve graft and both died of infectious complications;
one showed delayed, but complete, engraftment after this myeloablative
regimen. Two of the nine evaluable patients attained a clinical
complete remission (CR). The grafts from three patients were tested for
tumor contamination and contained no detectable clonal myeloma cells.
Larger quantities of purified cells may be required to resolve the
problem of delayed engraftment.
Blood, Vol. 91 No. 12 (June 15), 1998:
pp. 4489-4495
© 1998 by The American Society of Hematology.
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