Hisashi Oishi1, Stephen C Juvet1, Tereza Martinu1, Masaaki Sato2, Jeffrey A Medin3, Mingyao Liu1, Shaf Keshavjee4. 1. Latner Thoracic Surgery Research Laboratories, University Health Network, University of Toronto, Toronto, Ontario, Canada. 2. Latner Thoracic Surgery Research Laboratories, University Health Network, University of Toronto, Toronto, Ontario, Canada; Department of Thoracic Surgery, University of Tokyo, Tokyo, Japan. 3. Departments of Pediatrics and Biochemistry, Medical College of Wisconsin, Milwaukee, Wis. 4. Latner Thoracic Surgery Research Laboratories, University Health Network, University of Toronto, Toronto, Ontario, Canada. Electronic address: shaf.keshavjee@uhn.ca.
Abstract
OBJECTIVE: Our objective was to develop a rapid-onset and durable gene-delivery strategy that is applicable at the time of transplant to determine its effects on both acute rejection and chronic lung allograft fibrosis using a mouse orthotopic lung transplant model. METHODS: C57BL/6 mice received an orthotopic left lung transplant from syngeneic donors or C57BL/10 donors. By using syngeneic lung transplantation, we established a novel gene transfer protocol with lentivirus luciferase intrabronchial administration to the donor lung ex vivo before transplantation. This strategy was applied in allogeneic lung transplantation with lentivirus engineering expression of human interleukin-10 or lentivirus luciferase (control). RESULTS: Bioluminescent imaging revealed that the highest early transgene expression was achieved when lentivirus luciferase was administered both directly into the donor lung graft ex vivo before implantation and subsequently to the recipient in vivo daily on post-transplant days 1 to 4, compared with post-transplant in vivo administration only (days 0 to 4). Our previous work with adenoviral interleukin-10 gene therapy indicates that early interleukin-10 expression in the allograft is desirable. Therefore, we selected the combined protocol for human interleukin-10 encoding lentiviral vector therapy. In the allogeneic transplant setting, ex vivo and in vivo human interleukin-10 encoding lentiviral vector therapy reduced acute rejection grade (2.0 vs 3.0, P < .05) at day 28. The percentage of fibrotic obliterated airways was reduced in the human interleukin-10 encoding lentiviral vector-treated group (10.9% ± 7.7% vs 40.9% ± 9.3%, P < .05). CONCLUSIONS: Delivery of lentiviral interleukin-10 gene therapy, using a novel combined ex vivo and in vivo delivery strategy, significantly improves acute and chronic rejection in the mouse lung transplant model.
OBJECTIVE: Our objective was to develop a rapid-onset and durable gene-delivery strategy that is applicable at the time of transplant to determine its effects on both acute rejection and chronic lung allograft fibrosis using a mouse orthotopic lung transplant model. METHODS: C57BL/6 mice received an orthotopic left lung transplant from syngeneic donors or C57BL/10 donors. By using syngeneic lung transplantation, we established a novel gene transfer protocol with lentivirus luciferase intrabronchial administration to the donor lung ex vivo before transplantation. This strategy was applied in allogeneic lung transplantation with lentivirus engineering expression of humaninterleukin-10 or lentivirus luciferase (control). RESULTS: Bioluminescent imaging revealed that the highest early transgene expression was achieved when lentivirus luciferase was administered both directly into the donor lung graft ex vivo before implantation and subsequently to the recipient in vivo daily on post-transplant days 1 to 4, compared with post-transplant in vivo administration only (days 0 to 4). Our previous work with adenoviral interleukin-10 gene therapy indicates that early interleukin-10 expression in the allograft is desirable. Therefore, we selected the combined protocol for humaninterleukin-10 encoding lentiviral vector therapy. In the allogeneic transplant setting, ex vivo and in vivo humaninterleukin-10 encoding lentiviral vector therapy reduced acute rejection grade (2.0 vs 3.0, P < .05) at day 28. The percentage of fibrotic obliterated airways was reduced in the humaninterleukin-10 encoding lentiviral vector-treated group (10.9% ± 7.7% vs 40.9% ± 9.3%, P < .05). CONCLUSIONS: Delivery of lentiviral interleukin-10 gene therapy, using a novel combined ex vivo and in vivo delivery strategy, significantly improves acute and chronic rejection in the mouse lung transplant model.
Authors: Qimeng Gao; Isabel F DeLaura; Imran J Anwar; Samuel J Kesseli; Riley Kahan; Nader Abraham; Aravind Asokan; Andrew S Barbas; Matthew G Hartwig Journal: Front Immunol Date: 2022-07-01 Impact factor: 8.786
Authors: Xin Jin; Janne Kaes; Jan Van Slambrouck; Ilhan Inci; Stephan Arni; Vincent Geudens; Tobias Heigl; Yanina Jansen; Marianne S Carlon; Robin Vos; Dirk Van Raemdonck; Yi Zhang; Bart M Vanaudenaerde; Laurens J Ceulemans Journal: Cells Date: 2022-01-30 Impact factor: 6.600