Safety and efficacy of a lentiviral vector containing three anti-HIV genes--CCR5 ribozyme, tat-rev siRNA, and TAR decoy--in SCID-hu mouse-derived T cells.
Gene therapeutic strategies show promise in controlling human immunodeficiency virus (HIV) infection and in restoring immunological function. A number of efficacious anti-HIV gene constructs have been described so far, including small interfering RNAs (siRNAs), RNA decoys, transdominant proteins, and ribozymes, each with a different mode of action. However, as HIV is prone to generating escape mutants, the use of a single anti-HIV construct would not be adequate to afford long range-viral protection. On this basis, a combination of highly potent anti-HIV genes--namely, a short hairpin siRNA (shRNA) targeting rev and tat, a transactivation response (TAR) decoy, and a CCR5 ribozyme--have been inserted into a third-generation lentiviral vector. Our recent in vitro studies with this construct, Triple-R, established its efficacy in both T-cell lines and CD34 cell-derived macrophages. In this study, we have evaluated this combinatorial vector in vivo. Vector-transduced CD34 cells were injected into severe combined immunodeficiency (SCID)-hu mouse thy/liv grafts to determine their capacity to give rise to T cells. Our results show that phenotypically normal transgenic T cells are generated that are able to resist HIV-1 infection when challenged in vitro. These important attributes of this combinatorial vector show its promise as an excellent candidate for use in human clinical trials.
Anderson
Joseph
J
Department of Microbiology, Immunology and Pathology, Colorado State University, Fort, Collins, Colorado 80523, USA.
Li
Ming-Jie
MJ
Palmer
Brent
B
Remling
Leila
L
Li
Shirley
S
Yam
Priscilla
P
Yee
Jiing-Kuan
JK
Rossi
John
J
Zaia
John
J
Akkina
Ramesh
R
D000818:Animals; D018952:Antigens, CD34 / Q000235:genetics / Q000276:immunology; D005434:Flow Cytometry; D020868:Gene Silencing; D015757:Genes, rev / Q000235:genetics*; D015756:Genes, tat / Q000235:genetics*; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006678:HIV / Q000235:genetics*; D006801:Humans; D008264:Macrophages / Q000276:immunology / Q000378:metabolism; D051379:Mice; D016513:Mice, SCID; D016337:RNA, Catalytic / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics*; D019713:Receptors, CCR5 / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism* / Q000821:virology; D013950:Thymus Gland / Q000166:cytology / Q000276:immunology / Q000821:virology; D018183:Transplantation Chimera / Q000276:immunology
D018952:Antigens, CD34; D016337:RNA, Catalytic; D034741:RNA, Small Interfering; D019713:Receptors, CCR5; D049452:Green Fluorescent Proteins