title: High-efficiency transduction of rhesus hematopoietic repopulating cells by a modified HIV1-based lentiviral vector.
abstract: Human immunodeficiency virus type 1 (HIV1) vectors poorly transduce rhesus hematopoietic cells due to species-specific restriction factors, including the tripartite motif-containing 5 isoformα (TRIM5α) which targets the HIV1 capsid. We previously developed a chimeric HIV1 (χHIV) vector system wherein the vector genome is packaged with the simian immunodeficiency virus (SIV) capsid for efficient transduction of both rhesus and human CD34(+) cells. To evaluate whether χHIV vectors could efficiently transduce rhesus hematopoietic repopulating cells, we performed a competitive repopulation assay in rhesus macaques, in which half of the CD34(+) cells were transduced with standard SIV vectors and the other half with χHIV vectors. As compared with SIV vectors, χHIV vectors achieved higher vector integration, and the transgene expression rates were two- to threefold higher in granulocytes and red blood cells and equivalent in lymphocytes and platelets for 2 years. A recipient of χHIV vector-only transduced cells reached up to 40% of transgene expression rates in granulocytes and lymphocytes and 20% in red blood cells. Similar to HIV1 and SIV vectors, χHIV vector frequently integrated into gene regions, especially into introns. In summary, our χHIV vector demonstrated efficient transduction for rhesus long-term repopulating cells, comparable with SIV vectors. This χHIV vector should allow preclinical testing of HIV1-based therapeutic vectors in large animal models.
authors.affiliation: Molecular and Clinical Hematology Branch, National Heart Lung and Blood Institutes (NHLBI)/National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National Institutes of Health (NIH), Bethesda, Maryland, USA.
authors.forename: Naoya
authors.initials: N
authors.lastname: Uchida
authors.forename: Phillip W
authors.initials: PW
authors.lastname: Hargrove
authors.forename: Coen J
authors.initials: CJ
authors.lastname: Lap
authors.forename: Molly E
authors.initials: ME
authors.lastname: Evans
authors.forename: Oswald
authors.initials: O
authors.lastname: Phang
authors.forename: Aylin C
authors.initials: AC
authors.lastname: Bonifacino
authors.forename: Allen E
authors.initials: AE
authors.lastname: Krouse
authors.forename: Mark E
authors.initials: ME
authors.lastname: Metzger
authors.forename: Anh-Dao
authors.initials: AD
authors.lastname: Nguyen
authors.forename: Matthew M
authors.initials: MM
authors.lastname: Hsieh
authors.forename: Tyra G
authors.initials: TG
authors.lastname: Wolfsberg
authors.forename: Robert E
authors.initials: RE
authors.lastname: Donahue
authors.forename: Derek A
authors.initials: DA
authors.lastname: Persons
authors.forename: John F
authors.initials: JF
authors.lastname: Tisdale
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D015139:Blotting, Southern; D002213:Capsid / Q000378:metabolism; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D005822:Genetic Vectors / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics*; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells; D006801:Humans; D008253:Macaca mulatta; D060888:Real-Time Polymerase Chain Reaction; D015302:Simian Immunodeficiency Virus / Q000235:genetics; D014161:Transduction, Genetic; D019076:Transgenes
chemical_list: D018952:Antigens, CD34; D036022:Capsid Proteins