title: Preintegration HIV-1 inhibition by a combination lentiviral vector containing a chimeric TRIM5 alpha protein, a CCR5 shRNA, and a TAR decoy.
abstract: Human immunodeficiency virus (HIV) gene therapy offers a promising alternative approach to current antiretroviral treatments to inhibit HIV-1 infection. Various stages of the HIV life cycle including pre-entry, preintegration, and postintegration can be targeted by gene therapy to block viral infection and replication. By combining multiple highly potent anti-HIV transgenes in a single gene therapy vector, HIV-1 resistance can be achieved in transduced cells while prohibiting the generation of escape mutants. Here, we describe a combination lentiviral vector that encodes three highly effective anti-HIV genes functioning at separate stages of the viral life cycle including a CCR5 short hairpin RNA (shRNA) (pre-entry), a human/rhesus macaque chimeric TRIM5 alpha (postentry/preintegration), and a transactivation response element (TAR) decoy (postintegration). The major focus on designing this anti-HIV vector was to block productive infection of HIV-1 and to inhibit any formation of provirus that would maintain the viral reservoir. Upon viral challenge, potent preintegration inhibition of HIV-1 infection was achieved in combination vector-transduced cells in both cultured and primary CD34(+) hematopoietic progenitor cell (HPC)-derived macrophages. The generation of escape mutants was also blocked as evaluated by long-term culture of challenged cells. The ability of this combination anti-HIV lentiviral vector to prevent HIV-1 infection, in vitro, warrants further evaluation of its in vivo efficacy.
authors.affiliation: Stem Cell Program, Department of Internal Medicine, University of California-Davis, Sacramento, California, USA. joseph.anderson@ucdmc.ucdavis.edu
authors.forename: Joseph S
authors.initials: JS
authors.lastname: Anderson
authors.forename: John
authors.initials: J
authors.lastname: Javien
authors.forename: Jan A
authors.initials: JA
authors.lastname: Nolta
authors.forename: Gerhard
authors.initials: G
authors.lastname: Bauer
mesh_terms: D000818:Animals; D000090103:Antiviral Restriction Factors; D065100:CCR5 Receptor Antagonists; D002352:Carrier Proteins / Q000235:genetics*; D005434:Flow Cytometry; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage*; D006678:HIV / Q000235:genetics; D015658:HIV Infections / Q000235:genetics / Q000628:therapy* / Q000821:virology; D006412:Hematopoietic Stem Cells / Q000378:metabolism; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008253:Macaca mulatta; D008264:Macrophages / Q000378:metabolism; D011506:Proteins / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics*; D019713:Receptors, CCR5 / Q000235:genetics*; D014161:Transduction, Genetic; D019076:Transgenes / Q000502:physiology; D000070584:Tripartite Motif Proteins; D044767:Ubiquitin-Protein Ligases; D016662:Virus Integration; D054321:rev Gene Products, Human Immunodeficiency Virus / Q000037:antagonists & inhibitors / Q000502:physiology*; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000037:antagonists & inhibitors / Q000502:physiology
chemical_list: D000090103:Antiviral Restriction Factors; D065100:CCR5 Receptor Antagonists; D002352:Carrier Proteins; D011506:Proteins; D034741:RNA, Small Interfering; D019713:Receptors, CCR5; D000070584:Tripartite Motif Proteins; D054321:rev Gene Products, Human Immunodeficiency Virus; D054322:tat Gene Products, Human Immunodeficiency Virus; C477346:TRIM5 protein, human; C482587:TRIM5(alpha) protein, rhesus monkey; D044767:Ubiquitin-Protein Ligases
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