title: Isolation and transduction of CD34+ cells from small quantities of peripheral blood from HIV-1-infected patients not treated with hemopoietic growth factors.
abstract: A proposed hemopoietic stem cell gene therapy for treatment for HIV infection would involve transduction of CD34+ hemopoietic stem cells with vectors encoding anti-HIV constructs. Peripheral blood has proved to be a useful source of these hemopoietic stem cells and this study exploits this finding. Small quantities of peripheral blood were obtained from HIV-negative patients and HIV-positive patients who were and were not receiving hemopoietic growth factors (HGFs). CD34+ cells were obtained from these samples using a simple technique and scored for frequency of colony type. This demonstrated that HIV-negative patients had the highest frequency of colony-forming units (CFUs). HIV-positive patients not treated with HGFs had a lower frequency of CFUs, but the same colony type distribution as HIV-negative patients. HIV-positive patients treated with HGFs had the lowest frequency of CFUs, but their colony type distribution demonstrated that they had responded to treatment. CD34+ cells selected in this way were also transduced with the murine retroviral MFG vector using a technique that demonstrated transduction efficiencies ranging from 2% to 16% (median, 11.5%). This study simplifies the experimental requirements for development of a hemopoietic stem cell gene therapy for HIV infection and offers the possibility that longitudinal studies could be performed on peripheral blood CD34+ cells from HIV-positive or HIV-negative patients without the need for granulocyte colony-stimulating factor mobilization.
authors.affiliation: Department of Genito-Urinary Medicine and Communicable Diseases, Jefferiss Research Trust Laboratories, Imperial College School of Medicine at St. Mary's, London, UK.
authors.forename: G B
authors.initials: GB
authors.lastname: Adams
authors.forename: M
authors.initials: M
authors.lastname: McMullen
authors.forename: S
authors.initials: S
authors.lastname: Turner
authors.forename: I T
authors.initials: IT
authors.lastname: Olszak
authors.forename: D T
authors.initials: DT
authors.lastname: Scadden
authors.forename: M O
authors.initials: MO
authors.lastname: McClure
authors.forename: M C
authors.initials: MC
authors.lastname: Poznansky
mesh_terms: D000328:Adult; D018952:Antigens, CD34 / Q000097:blood*; D002452:Cell Count; D018929:Cell Culture Techniques; D002469:Cell Separation; D002472:Cell Transformation, Viral; D018920:Coculture Techniques; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D005434:Flow Cytometry; D015316:Genetic Therapy; D015658:HIV Infections / Q000097:blood* / Q000628:therapy; D015497:HIV-1 / Q000235:genetics; D016298:Hematopoietic Cell Growth Factors / Q000627:therapeutic use*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000276:immunology*; D006801:Humans; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D011533:Proviruses / Q000235:genetics
chemical_list: D018952:Antigens, CD34; D004279:DNA, Viral; D016298:Hematopoietic Cell Growth Factors