Clonal analysis of individual marrow-repopulating cells after experimental peripheral blood progenitor cell transplantation.
Methods to analyze the clonality of an adverse event in preclinical or clinical retroviral stem cell gene therapy protocols are needed. We analyzed the progeny of retrovirally transduced human peripheral blood progenitor cells (PBPCs) after transplantation and engraftment in immune-deficient mice. The integration site of the provirus serves as a unique tag of the individual transduced PBPC. A plasmid library of junctions between proviral and human genomic DNA was generated. We were able to detect individual transduced cell clones that amounted to 0.14%-0.0001% of chimeric bone marrow cells. This is the first report in which the contribution of individual marrow-repopulating cells to human hematopoiesis is directly quantified.
German Cancer Research Center, Heidelberg, Germany.
K Zsuzsanna
KZ
Nagy
Stephanie
S
Laufs
Bernhard
B
Gentner
Sonja
S
Naundorf
Klaus
K
Kuehlcke
Julian
J
Topaly
Eike C
EC
Buss
W Jens
WJ
Zeller
Stefan
S
Fruehauf
D000818:Animals; D018952:Antigens, CD34 / Q000097:blood*; D001854:Bone Marrow Cells / Q000166:cytology*; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D033581:Stem Cell Transplantation / Q000379:methods*; D014183:Transplantation, Heterologous / Q000276:immunology*
D018952:Antigens, CD34