title: Hematopoietic stem cell-based gene therapy for acquired immunodeficiency syndrome: efficient transduction and expression of RevM10 in myeloid cells in vivo and in vitro.
abstract: Gene delivery via the hematopoietic stem cell (HSC) offers an attractive means to introduce antiviral genes into both T cells and macrophages for acquired immunodeficiency syndrome (AIDS) gene therapy. An amphotropic retroviral vector encoding a bicistronic gene coexpressing RevM10 and the murine CD8alpha' chain (lyt2) was developed to transduce HSC/progenitor cells. After transduction of CD34+ cells isolated from human umbilical cord blood, the lyt2 molecule detected by flow cytometry was used to monitor the level of gene transduction and expression and to enrich RevM10-expressing cells by cell sorting without drug selection. Using this quantitative method, high levels of gene transduction and expression (around 20%) were achieved by high-speed centrifugation of CD34+ cells with the retroviral supernatant (spinoculation). After reconstitution of human bone marrow implanted in SCID mice (SCID-hu bone) with the transduced HSC/progenitor cells, a significant number of donor-derived CD14+ bone marrow cells were found to express the RevM10/lyt2 gene. Finally, replication of a macrophage-tropic human immunodeficiency virus-type 1 (HIV-1) isolate was greatly inhibited in the lyt2+/CD14+ cells differentiated from transduced CD34+ cells after the enrichment of lyt2+ population. Thus, the RevM10 gene did not appear to inhibit the differentiation of HSC/progneitor cells into monocytes/macrophages. The level of retrovirus-mediated RevM10 expression in monocytes/macrophages derived from transduced HSCs is sufficient to suppress HIV-1 replication.
authors.affiliation: Progenesys Program, SyStemix, Inc, Palo Alto, CA 94304, USA.
authors.forename: L
authors.initials: L
authors.lastname: Su
authors.forename: R
authors.initials: R
authors.lastname: Lee
authors.forename: M
authors.initials: M
authors.lastname: Bonyhadi
authors.forename: H
authors.initials: H
authors.lastname: Matsuzaki
authors.forename: S
authors.initials: S
authors.lastname: Forestell
authors.forename: S
authors.initials: S
authors.lastname: Escaich
authors.forename: E
authors.initials: E
authors.lastname: Böhnlein
authors.forename: H
authors.initials: H
authors.lastname: Kaneshima
mesh_terms: D000163:Acquired Immunodeficiency Syndrome / Q000628:therapy*; D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D005312:Fetal Blood / Q000166:cytology; D015694:Gene Products, rev / Q000172:deficiency / Q000235:genetics / Q000502:physiology; D017930:Genes, Reporter; D015757:Genes, rev; D015316:Genetic Therapy; D015497:HIV-1 / Q000502:physiology; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000821:virology; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D014162:Transfection; D054321:rev Gene Products, Human Immunodeficiency Virus
chemical_list: D015694:Gene Products, rev; D054321:rev Gene Products, Human Immunodeficiency Virus