Hematopoietic potential and retroviral transduction of CD34+ Thy-1+ peripheral blood stem cells from asymptomatic human immunodeficiency virus type-1-infected individuals mobilized with granulocyte colony-stimulating factor.
The potential of hematopoietic stem cells (HSCs) from human immunodeficiency virus type-1 (HIV-1)-infected individuals, eg, self-renewal and multilineage differentiative capacity, might be perturbed due to the underlying disease. In this study, we assessed the HSC activity in the CD34+ Thy-1+ cell population of peripheral blood stem cells (PBSCs) of three asymptomatic HIV-1-infected individuals after granulocyte colony-stimulating factor (G-CSF; 10 microg/kg/d) mobilization. On day 4 of G-CSF treatment, 0.8% to 1% of the total blood mononuclear cells were CD34+. Leukapheresis followed by a two-step cell isolation process yielded a CD34+ Thy-1+ cell population of high purity (76% to 92% CD34+ Thy-1+ cells). This cell population showed no evidence of HIV-1-containing cells based on a semiquantitative HIV-1 DNA polymerase chain reaction. Furthermore, the purified cells showed normal hematopoietic potential in in vitro clonogenic assays. Successful gene transfer into committed progenitor cells (colony-forming units-cells) and more primitive stem/progenitor cells (long-term culture colony-forming cells) could be shown after amphotropic retroviral transduction. These data provide evidence that the CD34+ Thy-1+ stem cell compartment can be mobilized and enriched in early stage HIV-1-infected patients. Furthermore, successful transduction of this cell population as a prerequisite for stem cell-based clinical gene therapy protocols was demonstrated.
Junker
U
U
Progenesys Program, Systemix Inc, Palo Alto, CA 94304, USA.
Moon
J J
JJ
Kalfoglou
C S
CS
Sniecinski
I
I
Forman
S J
SJ
Zaia
J A
JA
Kaneshima
H
H
Böhnlein
E
E
D000328:Adult; D018952:Antigens, CD34 / Q000032:analysis; D001772:Blood Cell Count; D002469:Cell Separation; D004279:DNA, Viral / Q000097:blood; D005240:Feasibility Studies; D005260:Female; D005434:Flow Cytometry; D005822:Genetic Vectors; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology*; D016655:HIV Core Protein p24 / Q000097:blood; D015658:HIV Infections / Q000097:blood*; D006410:Hematopoiesis / Q000187:drug effects*; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000187:drug effects* / Q000821:virology; D006801:Humans; D007937:Leukapheresis; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D012190:Retroviridae / Q000235:genetics*; D018800:Thy-1 Antigens / Q000032:analysis; D014766:Viremia / Q000097:blood
D018952:Antigens, CD34; D004279:DNA, Viral; D016655:HIV Core Protein p24; D018800:Thy-1 Antigens; D016179:Granulocyte Colony-Stimulating Factor