Multilineage long-term engraftment potential of drug-resistant hematopoietic progenitors.
Peripheral blood progenitor cells (PBPCs) are increasingly used instead of bone marrow for autologous or allogeneic transplantation. In this study PBPCs mobilized in cancer patients by chemotherapy and granulocyte-colony stimulating factor were collected by apheresis and first enriched by immunoaffinity removal of lineage positive cells. When these cells were exposed to both cyclophosphamide and taxol or cultured for 7 days in the presence of 5-fluorouracil, stem cell factor, and interleukin-3, 88% to 93% of the enriched PBPCs were killed and short-term clonogenic capacity in methylcellulose assays was lost, but week-5 cobblestone area-forming cell (CAFC) enrichment was higher than 10-fold in comparison to enriched PBPCs and higher than 700-fold in comparison to unmanipulated apheresis cells. After drug exposure, most of the progenitors displayed a CD34+, CD38-, multidrug-resistance (MDR+), Rhodamine 123 low, Hoechst 33342 low phenotype, and as few as 180 of these drug-resistant cells were able to generate a stable multilineage human hematopoiesis in sublethally irradiated immunodeficient mice. In these animals, the level of human hematopoietic engraftment was significantly increased by cotransplantation of irradiated cells from the human L87/4 stromal cell line. These observations are consistent with the functional isolation of a population of very early hematopoietic progenitors and might help to design new protocols for the removal of neoplastic cells from autografts.
Bertolini
F
F
Division of Medical Oncology, IRCCS Maugeri Foundation, Pavia Medical Center, Italy.
Battaglia
M
M
Lanza
A
A
Gibelli
N
N
Palermo
B
B
Pavesi
L
L
Caprotti
M
M
Robustelli della Cuna
G
G
D000818:Animals; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000494:pharmacology*; D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D001781:Blood Component Removal; D002478:Cells, Cultured; D003520:Cyclophosphamide / Q000031:analogs & derivatives / Q000494:pharmacology; D004351:Drug Resistance; D005472:Fluorouracil / Q000494:pharmacology; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000187:drug effects*; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology; D008562:Membrane Glycoproteins / Q000032:analysis / Q000235:genetics; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D017239:Paclitaxel / Q000494:pharmacology; D010641:Phenotype; D011956:Receptors, Cell Surface; D019089:Stem Cell Factor / Q000494:pharmacology; D019172:Transplantation Conditioning; D014960:X Chromosome
D000964:Antimetabolites, Antineoplastic; D000970:Antineoplastic Agents; D018906:Antineoplastic Agents, Alkylating; D000972:Antineoplastic Agents, Phytogenic; D007377:Interleukin-3; D008562:Membrane Glycoproteins; C097871:PLXNA3 protein, human; D011956:Receptors, Cell Surface; D019089:Stem Cell Factor; C048341:mafosfamide; D003520:Cyclophosphamide; D017239:Paclitaxel; D005472:Fluorouracil