title: In vivo safety and persistence of endoribonuclease gene-transduced CD4+ T cells in cynomolgus macaques for HIV-1 gene therapy model.
abstract: BACKGROUND
  MazF is an endoribonuclease encoded by Escherichia coli that specifically cleaves the ACA sequence of mRNA. In our previous report, conditional expression of MazF in the HIV-1 LTR rendered CD4+ T lymphocytes resistant to HIV-1 replication. In this study, we examined the in vivo safety and persistence of MazF-transduced cynomolgus macaque CD4+ T cells infused into autologous monkeys.
  
  
  RESULTS
  The in vivo persistence of the gene-modified CD4+ T cells in the peripheral blood was monitored for more than half a year using quantitative real-time PCR and flow cytometry, followed by experimental autopsy in order to examine the safety and distribution pattern of the infused cells in several organs. Although the levels of the MazF-transduced CD4+ T cells gradually decreased in the peripheral blood, they were clearly detected throughout the experimental period. Moreover, the infused cells were detected in the distal lymphoid tissues, such as several lymph nodes and the spleen. Histopathological analyses of tissues revealed that there were no lesions related to the infused gene modified cells. Antibodies against MazF were not detected. These data suggest the safety and the low immunogenicity of MazF-transduced CD4+ T cells. Finally, gene modified cells harvested from the monkey more than half a year post-infusion suppressed the replication of SHIV 89.6P.
  
  
  CONCLUSIONS
  The long-term persistence, safety and continuous HIV replication resistance of the mazF gene-modified CD4+ T cells in the non-human primate model suggests that autologous transplantation of mazF gene-modified cells is an attractive strategy for HIV gene therapy.
authors.affiliation: Center for Cell and Gene Therapy, Takara Bio Inc, Otsu, Shiga, Japan. chonoh@takara-bio.co.jp
authors.forename: Hideto
authors.initials: H
authors.lastname: Chono
authors.forename: Naoki
authors.initials: N
authors.lastname: Saito
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Tsuda
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Shibata
authors.forename: Naohide
authors.initials: N
authors.lastname: Ageyama
authors.forename: Keiji
authors.initials: K
authors.lastname: Terao
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Yasutomi
authors.forename: Junichi
authors.initials: J
authors.lastname: Mineno
authors.forename: Ikunoshin
authors.initials: I
authors.lastname: Kato
mesh_terms: D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism* / Q000637:transplantation; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004195:Disease Models, Animal; D004720:Endonucleases / Q000235:genetics / Q000378:metabolism*; D004722:Endoribonucleases / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005434:Flow Cytometry; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D015658:HIV Infections / Q000235:genetics / Q000276:immunology / Q000628:therapy*; D015497:HIV-1 / Q000276:immunology; D006801:Humans; D008198:Lymph Nodes / Q000276:immunology / Q000378:metabolism; D008252:Macaca fascicularis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013154:Spleen / Q000276:immunology / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D014182:Transplantation, Autologous; D016896:Treatment Outcome; D014779:Virus Replication / Q000276:immunology
chemical_list: D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C422955:MazF protein, E coli; D004720:Endonucleases; D004722:Endoribonucleases