title: Efficient transduction of human hematopoietic repopulating cells with a chimeric HIV1-based vector including SIV capsid.
abstract: Innate immune factors, such as TRIM5α and cyclophilin A (CypA), act as a major restriction factor of retroviral infection among species. When HIV1 infects human cells, HIV1 capsid binds to human CypA to escape from human TRIM5α restriction. However, in rhesus cells, the mismatch between HIV1 capsid and rhesus CypA is recognized by rhesus TRIM5α to reduce HIV1 infectivity through proteasomal degradation. To circumvent this block, we previously developed a chimeric HIV1 vector (χHIV) that substituted HIV1 capsid with SIV capsid, and it significantly increased transduction efficiency for nonhuman primate cells. In this study, we evaluated whether the χHIV vector efficiently transduces human cells, and the transduction efficiency might increase by a CypA inhibitor (cyclosporine) and a proteasome inhibitor (MG132). The χHIV vector could transduce human CD34⁺ cells, as efficiently as the HIV1 vector, in vitro and in xenograft mice, even in the mismatch between SIV capsid and human CypA. Cyclosporine decreased transduction efficiency with the HIV1 vector, whereas it slightly increased transduction efficiency with the χHIV vector in human CD34⁺ cells. MG132 increased transduction efficiency with both χHIV and HIV1 vectors in the same manner. However, MG132 was toxic to human CD34⁺ cells at high concentrations, and both drugs had a small range of effective dosage. These findings demonstrate that both χHIV and HIV1 vectors have similar transduction efficiency for human hematopoietic repopulating cells, suggesting that the χHIV vector escapes from TRIM5α restriction, which is independent of human CypA.
authors.affiliation: Molecular and Clinical Hematology Branch, National Heart Lung and Blood Institute-National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Naoya
authors.initials: N
authors.lastname: Uchida
authors.forename: Matthew M
authors.initials: MM
authors.lastname: Hsieh
authors.forename: Kareem N
authors.initials: KN
authors.lastname: Washington
authors.forename: John F
authors.initials: JF
authors.lastname: Tisdale
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000235:genetics / Q000378:metabolism; D000090103:Antiviral Restriction Factors; D002213:Capsid; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics; D021984:Cyclophilin A / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics; D016572:Cyclosporine / Q000494:pharmacology; D005260:Female; D005822:Genetic Vectors / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology; D007976:Leupeptins / Q000494:pharmacology; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism; D061988:Proteasome Inhibitors / Q000494:pharmacology; D015302:Simian Immunodeficiency Virus / Q000235:genetics*; D014161:Transduction, Genetic; D000070584:Tripartite Motif Proteins; D044767:Ubiquitin-Protein Ligases
chemical_list: D018952:Antigens, CD34; D000090103:Antiviral Restriction Factors; D002352:Carrier Proteins; D007166:Immunosuppressive Agents; D007976:Leupeptins; D061988:Proteasome Inhibitors; D000070584:Tripartite Motif Proteins; D016572:Cyclosporine; C477346:TRIM5 protein, human; D044767:Ubiquitin-Protein Ligases; D046988:Proteasome Endopeptidase Complex; D021984:Cyclophilin A; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde