title: Targeting lentiviral vector expression to hepatocytes limits transgene-specific immune response and establishes long-term expression of human antihemophilic factor IX in mice.
abstract: Stable gene replacement by in vivo administration of lentiviral vectors (LVs) has therapeutic potential for metabolic disorders and other systemic diseases. We studied the expression of intracellular and secreted proteins by LVs in immunocompetent mice. Liver, spleen, and bone marrow cells were efficiently transduced. However, transgene expression, driven by a ubiquitous promoter, was limited by transgene-specific cellular and humoral immune responses, leading to the clearance of transduced cells. After green fluorescent protein (GFP) gene transfer, the liver showed infiltration of CD8(+) cytotoxic T cells, and GFP-specific CD8(+) T cells were isolated from the spleen. After human factor IX (hF.IX) gene transfer, anti-hF.IX antibodies were induced. These immune responses were not detected in mice injected with heat-inactivated or genome-lacking LVs or in GFP-transgenic mice, indicating that they were specifically triggered by transgene expression in vivo. Intriguingly, selective targeting of LV expression to hepatocytes limited the immune responses to the transgenes. By this approach, high levels of hF.IX, potentially in the therapeutic range, were reached and maintained long term in immunocompetent mice, without inducing antibody formation. These results prompt further studies in relevant animal models to explore the potential of in vivo LV administration for the gene therapy of hemophilias and other liver-based diseases.
authors.affiliation: Laboratory of Gene Transfer and Therapy, Institute for Cancer Research and Treatment (IRCC), University of Torino, Torino, Italy.
authors.forename: Antonia
authors.initials: A
authors.lastname: Follenzi
authors.forename: Manuela
authors.initials: M
authors.lastname: Battaglia
authors.forename: Angelo
authors.initials: A
authors.lastname: Lombardo
authors.forename: Andrea
authors.initials: A
authors.lastname: Annoni
authors.forename: Maria Grazia
authors.initials: MG
authors.lastname: Roncarolo
authors.forename: Luigi
authors.initials: L
authors.lastname: Naldini
mesh_terms: D000818:Animals; D005164:Factor IX / Q000008:administration & dosage* / Q000235:genetics / Q000276:immunology; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000627:therapeutic use*; D049452:Green Fluorescent Proteins; D022781:Hepatocytes / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007518:Isoantibodies; D016086:Lentivirus / Q000235:genetics; D008099:Liver / Q000276:immunology; D008164:Luminescent Proteins; D051379:Mice; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014161:Transduction, Genetic; D019076:Transgenes / Q000276:immunology
chemical_list: D007518:Isoantibodies; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D005164:Factor IX