title: Local long-term expression of lentivirally delivered IL-10 in the lung attenuates obliteration of intrapulmonary allograft airways.
abstract: Obliterative bronchiolitis (OB) is a form of chronic rejection after lung transplantation. Lentiviral vectors (LVs) facilitate long-term gene transduction in many tissues and organs. We hypothesized that lentiviral gene transfer of interleukin (IL)-10, a potent immune-modulating cytokine, to the lung could modulate the alloimmune responses in the lung after transplantation. C57BL6 mice received LVs encoding luciferase, enhanced green fluorescent protein (eGFP), or human IL-10 (huIL-10) through airways and underwent repeated bioluminescent imaging, immunofluorescence imaging, or ELISA of lung tissues, respectively. Luciferase activities peaked at day 7 and were stable after day 28 to over 15 months. eGFP staining demonstrated LV-mediated gene transduction mainly in alveolar macrophages. LV-huIL-10 delivery resulted in stable long-term expression of huIL-10 in the lung tissue (average 3.66 pg/mg at 1 year). Intrapulmonary allograft tracheal transplantation (BALBc→C57BL6) was used as a model of OB. LV-huIL-10 or LV-eGFP were delivered 7 days before transplantation and compared with no LV-transfection group. Allograft airways at day 28 were almost completely obliterated in all the groups. However, at day 42, allograft airways treated with LV-huIL-10 showed a spectrum of attenuation in airway fibrosis ranging from complete obliteration through bubble-like partial opening to complete patency with epithelial coverage in association with a significantly reduced obliteration ratio compared with the other groups (p<0.05). In conclusion, lentivirus-mediated gene transduction is useful in achieving long-term transgene expression in the lung. Long-term IL-10 expression has the potential to attenuate allograft airway obliteration. LV-mediated gene therapy could be a useful strategy to prevent or treat OB after lung transplantation.
authors.affiliation: Latner Thoracic Surgery Research Laboratories, University Health Network, University of Toronto, Toronto, Ontario M5G 2C4, Canada.
authors.forename: Shin
authors.initials: S
authors.lastname: Hirayama
authors.forename: Masaaki
authors.initials: M
authors.lastname: Sato
authors.forename: Mingyao
authors.initials: M
authors.lastname: Liu
authors.forename: Severine
authors.initials: S
authors.lastname: Loisel-Meyer
authors.forename: Jonathan C
authors.initials: JC
authors.lastname: Yeung
authors.forename: Dirk
authors.initials: D
authors.lastname: Wagnetz
authors.forename: Marcelo
authors.initials: M
authors.lastname: Cypel
authors.forename: Guan
authors.initials: G
authors.lastname: Zehong
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Medin
authors.forename: Shaf
authors.initials: S
authors.lastname: Keshavjee
mesh_terms: D000818:Animals; D001989:Bronchiolitis Obliterans / Q000235:genetics* / Q000473:pathology / Q000628:therapy*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D016753:Interleukin-10 / Q000235:genetics* / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics*; D008168:Lung / Q000473:pathology; D016040:Lung Transplantation; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D014184:Transplantation, Homologous
chemical_list: C485184:enhanced green fluorescent protein; D016753:Interleukin-10; D049452:Green Fluorescent Proteins