RevM10-expressing T cells derived in vivo from transduced human hematopoietic stem-progenitor cells inhibit human immunodeficiency virus replication.
A key feature of the pathogenesis of human immunodeficiency virus type 1 (HIV-1) infection is the gradual loss of CD4-positive T cells. A number of gene therapy strategies have been designed with the intent of inhibiting HIV replication in mature T cells. As T cells are products of hematolymphoid differentiation, insertion of antiviral genes into hematopoietic stem cells could serve as a vehicle to confer long-term protection in progeny T cells derived from transduced stem cells. One such "cellular immunization" strategy utilizes the gene coding for the HIV-1 rev trans-dominant mutant protein RevM10 which has been demonstrated to inhibit HIV-1 replication in T-cell lines and in primary T cells. In this study, we used a Moloney murine leukemia virus-based retrovirus encoding a bicistronic message coexpressing RevM10 and the murine CD8-alpha' chain (Lyt2). This vector allows rapid selection of transgene-expressing cells as well as quantitation of transgene expression. We demonstrate that RevM10-transduced CD34-enriched hematopoietic progenitor-stem cells (HPSC) isolated from human umbilical cord blood or from granulocyte colony-stimulating factor-mobilized peripheral blood can give rise to mature thymocytes in the SCID-hu thymus/liver mouse model. The phenotypic distribution of HPSC-derived thymocytes is normal, and expression of the transgene can be detected by flow cytometric analysis. Moreover, we demonstrate that RevM10 can inhibit HIV replication in T cells derived from transduced HPSC after expansion in vitro. This is the first demonstration of anti-HIV efficacy in T cells derived from transduced human HPSC.
Bonyhadi
M L
ML
Progenesys Program, SyStemix, Inc., Palo Alto, California 94304, USA. mbonyhad@stem.com
Moss
K
K
Voytovich
A
A
Auten
J
J
Kalfoglou
C
C
Plavec
I
I
Forestell
S
S
Su
L
L
Böhnlein
E
E
Kaneshima
H
H
D000818:Animals; D019380:Anti-HIV Agents / Q000008:administration & dosage; D000950:Antigens, Ly / Q000235:genetics; D015496:CD4-Positive T-Lymphocytes / Q000382:microbiology*; D015694:Gene Products, rev / Q000008:administration & dosage / Q000235:genetics; D005799:Genes, Dominant; D015757:Genes, rev; D015316:Genetic Therapy / Q000379:methods; D015497:HIV-1 / Q000254:growth & development*; D006412:Hematopoietic Stem Cells / Q000382:microbiology*; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D008822:Mice, Transgenic; D008979:Moloney murine leukemia virus; D014161:Transduction, Genetic; D014779:Virus Replication; D054321:rev Gene Products, Human Immunodeficiency Virus
D019380:Anti-HIV Agents; D000950:Antigens, Ly; D015694:Gene Products, rev; D054321:rev Gene Products, Human Immunodeficiency Virus