Inhibition of HIV-1 by lentiviral vector-transduced siRNAs in T lymphocytes differentiated in SCID-hu mice and CD34+ progenitor cell-derived macrophages.
The phenomenon of RNA interference mediated by small interfering RNAs (siRNAs) is a potent gene-silencing mechanism. A number of recent studies demonstrated inhibition of HIV-1 replication in cultured cells using this approach. To make further progress and harness this technology for HIV-1 gene therapy in a stem cell setting, in vivo studies using primary hematopoietic cells are needed. Using an HIV-based lentiviral vector we introduced an anti-Rev siRNA construct into CD34(+) hematopoietic progenitor cells. The siRNA-transduced progenitor cells were allowed to mature into macrophages in vitro and T cells in vivo in SCID-hu mouse thy/liv grafts. Phenotypically normal T cells and macrophages displaying characteristic surface markers were obtained. In vitro HIV-1 challenge of the siRNA-expressing macrophages and T cells with macrophage-tropic and T-cell-tropic HIV-1, respectively, showed marked viral resistance. These experiments demonstrate the utility of siRNAs delivered into hematopoietic stem cells via lentiviral vectors for future in vivo applications.
Banerjea
Akhil
A
Department of Microbiology, Immunology and Pathology, Colorado State University, 1619 Campus Drive, Fort Collins, Colorado 80523, USA.
Li
Ming-Jie
MJ
Bauer
Gerhard
G
Remling
Leila
L
Lee
Nan-Sook
NS
Rossi
John
J
Akkina
Ramesh
R
D000818:Animals; D018952:Antigens, CD34 / Q000096:biosynthesis*; D002454:Cell Differentiation; D002469:Cell Separation; D003114:Colony-Forming Units Assay; D005434:Flow Cytometry; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D015658:HIV Infections / Q000628:therapy*; D015497:HIV-1 / Q000378:metabolism*; D006412:Hematopoietic Stem Cells / Q000378:metabolism; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008099:Liver / Q000196:embryology; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D016513:Mice, SCID; D008957:Models, Genetic; D010641:Phenotype; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D034741:RNA, Small Interfering / Q000235:genetics*; D013601:T-Lymphocytes / Q000378:metabolism*; D013950:Thymus Gland / Q000166:cytology; D013997:Time Factors; D014158:Transcription, Genetic
D018952:Antigens, CD34; D034741:RNA, Small Interfering