title: Efficient marking of human cells with rapid but transient repopulating activity in autografted recipients.
abstract: Short-term hematopoietic reconstituting cells have been identified in mice, nonhuman primates, and among human cells that engraft xenogeneic hosts. We now present clonal marking data demonstrating a rapid but unsustained contribution of cultured human autografts to the initial phase of hematologic recovery in myeloablated patients. Three patients received transplants of granulocyte colony-stimulating factor-mobilized autologous peripheral blood (PB) cells, of which a portion (8%-25% of the CD34+ cells) had been incubated in vitro with growth factors (5 days) and clinical grade LN retrovirus (3-5 days). More than 9% of the clonogenic and long-term culture-initiating cells harvested were transduced. Semiquantitative and linear amplification-mediated polymerase chain reaction analyses of serial PB samples showed that marked white blood cells appeared in all 3 patients within 11 days and transiently constituted up to 0.1% to 1% of those produced in the first month. However, within another 2 to 9 months, marked cells had permanently decreased to very low levels. Analysis of more than 50 vector insertion sites showed none of the clones detected in the first month were active later. Eighty percent of inserts were located within or near genes, 2 near CXCR4. These findings provide direct evidence of cells with rapid but transient repopulating activity in patients and demonstrate their efficient transduction in vitro.
authors.affiliation: Department of Internal Medicine I, Albert-Ludwigs-University, Freiburg, Germany.
authors.forename: Hanno
authors.initials: H
authors.lastname: Glimm
authors.forename: Manfred
authors.initials: M
authors.lastname: Schmidt
authors.forename: Marlene
authors.initials: M
authors.lastname: Fischer
authors.forename: Kerstin
authors.initials: K
authors.lastname: Schwarzwaelder
authors.forename: Manuela
authors.initials: M
authors.lastname: Wissler
authors.forename: Silke
authors.initials: S
authors.lastname: Klingenberg
authors.forename: Claudia
authors.initials: C
authors.lastname: Prinz
authors.forename: Cornelius F
authors.initials: CF
authors.lastname: Waller
authors.forename: Winand
authors.initials: W
authors.lastname: Lange
authors.forename: Connie J
authors.initials: CJ
authors.lastname: Eaves
authors.forename: Christof
authors.initials: C
authors.lastname: von Kalle
mesh_terms: D015415:Biomarkers; D002478:Cells, Cultured; D002999:Clone Cells; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000502:physiology*; D006801:Humans; D007700:Kinetics; D007962:Leukocytes; D036102:Peripheral Blood Stem Cell Transplantation; D012038:Regeneration; D012190:Retroviridae; D014161:Transduction, Genetic; D014182:Transplantation, Autologous; D016662:Virus Integration
chemical_list: D015415:Biomarkers