Inhibition of simian/human immunodeficiency virus replication in CD4+ T cells derived from lentiviral-transduced CD34+ hematopoietic cells.
We examined the ability of a HIV-1-based vector (VRX494) encoding a 937-bp antisense HIV-1 envelope sequence to inhibit the replication of chimeric SIV/HIV-1 viruses encoding the HIV-1 envelope. Challenge of VRX494-transduced CEMx174 cells resulted in potent inhibition of HIV-1 and several SHIV strains. To evaluate the potential efficacy of the VRX494 vector for stem cell gene therapy, rhesus CD34(+) bone marrow cells were transduced with VRX494 and then cultured on thymus stroma to induce T cell differentiation. Transduction conditions for CD34(+) cells were optimized to yield high transduction efficiency with minimal effective multiplicity of infection. Purified CD4(+) GFP(+) T cells derived from VRX494-transduced CD34(+) cells strongly inhibited SHIV HXBC2P 3.2 and SHIV 89.6P replication compared to controls. Southern blot analysis of VRX494-transduced T cell clones revealed a subset of cells with multiple proviral copies per cell. Expression of GFP and the antisense inhibitor in VRX494-transduced cells was upregulated by Tat. Analysis of HIV-1 envelope sequences in VRX494-transduced cells revealed modifications consistent with those mediated by double-stranded RNA-dependent adenosine deaminase. These results indicate that the macaque/SHIV model should serve as a useful preclinical model to evaluate this lentiviral vector expressing an HIV-1 antisense inhibitor for stem cell gene therapy for AIDS.
Braun
Stephen E
SE
Division of Immunology, New England Primate Research Center, Harvard Medical School, One Pine Hill Drive, Southborough, MA 01772, USA.
Wong
Fay Eng
FE
Connole
Michelle
M
Qiu
Gang
G
Lee
Lorrin
L
Gillis
Jackie
J
Lu
Xiaobin
X
Humeau
Laurent
L
Slepushkin
Vladimir
V
Binder
Gwendolyn K
GK
Dropulic
Boro
B
Johnson
R Paul
RP
D000243:Adenosine Deaminase / Q000378:metabolism; D000818:Animals; D018952:Antigens, CD34 / Q000096:biosynthesis* / Q000235:genetics; D015139:Blotting, Southern; D001854:Bone Marrow Cells / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D015686:Gene Products, env / Q000378:metabolism; D015694:Gene Products, rev / Q000378:metabolism; D015696:Gene Products, tat / Q000378:metabolism; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics*; D006412:Hematopoietic Stem Cells / Q000378:metabolism* / Q000821:virology; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008253:Macaca mulatta; D008957:Models, Genetic; D016376:Oligonucleotides, Antisense / Q000737:chemistry; D012313:RNA / Q000737:chemistry; D012190:Retroviridae / Q000235:genetics; D015302:Simian Immunodeficiency Virus / Q000235:genetics*; D013234:Stem Cells / Q000378:metabolism; D013601:T-Lymphocytes / Q000378:metabolism; D015854:Up-Regulation; D014779:Virus Replication; D054321:rev Gene Products, Human Immunodeficiency Virus; D054322:tat Gene Products, Human Immunodeficiency Virus
D018952:Antigens, CD34; D015686:Gene Products, env; D015694:Gene Products, rev; D015696:Gene Products, tat; D016376:Oligonucleotides, Antisense; D054321:rev Gene Products, Human Immunodeficiency Virus; D054322:tat Gene Products, Human Immunodeficiency Virus; D049452:Green Fluorescent Proteins; D012313:RNA; D000243:Adenosine Deaminase