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

• Published in: Blood 1997-06, Vol.89 (12), p.4299-4306
• Main Authors: Junker, Uwe, Moon, James J., Kalfoglou, Creton S., Sniecinski, Irena, Forman, Stephen J., Zaia, John A., Kaneshima, Hideto, Böhnlein, Ernst
• Format: Article
• Language: English
• Subjects: Adult; AIDS/HIV; Antigens, CD34 - analysis; Biological and medical sciences; Biotechnology; Blood Cell Count; Cell Separation; DNA, Viral - blood; Feasibility Studies; Female; Flow Cytometry; Fundamental and applied biological sciences. Psychology; Gene therapy; Genetic Vectors; Granulocyte Colony-Stimulating Factor - pharmacology; Health. Pharmaceutical industry; Hematopoiesis - drug effects; Hematopoietic Stem Cell Transplantation; Hematopoietic Stem Cells - drug effects; Hematopoietic Stem Cells - virology; HIV Core Protein p24 - blood; HIV Infections - blood; Humans; Immunodeficiencies; Immunodeficiencies. Immunoglobulinopathies; Immunopathology; Industrial applications and implications. Economical aspects; Leukapheresis; Male; Medical sciences; Middle Aged; Polymerase Chain Reaction; Retroviridae - genetics; Thy-1 Antigens - analysis; Viremia - blood
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood.V89.12.4299

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 μg/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.