Feasibility of baculovirus-mediated reporter gene delivery for efficient monitoring of islet transplantation in vivo |
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| Institution: | 1. Department of Nuclear Medicine, Ruijin Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200025, China;2. Institute of Endocrinology and Metabolism, Ruijin Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200025, China;1. Division of Cancer Research, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia;2. Division of Cancer Medicine, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia;3. Centre for Cancer Imaging, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia;4. Millennium Pharmaceuticals, Cambridge, MA;5. Takeda California, San Diego, CA;6. Department of Medicine, St Vincent’s Hospital, The University of Melbourne, Parkville, Victoria, Australia;7. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia;1. Division of Thoracic Surgery, Department of Surgery, Keio University School of Medicine, Tokyo, Japan;2. Division of Thoracic Surgery, Department of Surgery, Jikei University School of Medicine, Tokyo, Japan;3. Department of Pathology, Keio University School of Medicine, Tokyo, Japan;1. KU Leuven, Laboratory of Radiopharmacy, Leuven, Belgium;2. KU Leuven, Department of Imaging and Pathology, Leuven, Belgium and Nuclear Medicine, UZ Leuven;3. KU Leuven, Departement of Cardiovascular Sciences, Leuven, Belgium;4. C.R.E.A.Te Translational Sciences, Janssen Pharmaceutica, R&D, Beerse, Belgium |
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| Abstract: | ObjectiveThe objective of this study was to explore the feasibility of baculovirus vector-mediated sodium iodide symporter (NIS) gene delivery to monitor islet transplantation.MethodsBaculovirus vectors expressing green fluorescent protein (GFP) or NIS (Bac-GFP and Bac-NIS) were established using the Bac-to-Bac baculovirus expression system. The GFP expression of Bac-GFP-infected rat islets was observed in vitro by fluorescence microscopy. Iodine uptake and inhibition of iodine uptake by NaClO4 in Bac-NIS-infected islets were dynamically monitored in vitro. Bac-GFP- or Bac-NIS-infected islets were implanted into the left axillary cavity of NOD-SCID mice, and fluorescence imaging and 125I NanoSPECT/CT imaging were subsequently performed in vivo.ResultsBac-GFP efficiently infected rat islets (over 95% infected at MOI = 40), and the expression of GFP lasted approximately two weeks. NaClO4 could inhibit iodine uptake by Bac-NIS-infected islets. In vivo imaging revealed that the fluorescence intensity of the transplant sites in Bac-GFP-infected groups was significantly higher than in the non-infected group. Grafts could be clearly observed by 125I NanoSPECT/CT imaging for up to 8 h.ConclusionBaculovirus vectors are powerful vehicles for studying rat islets in gene delivery. It is feasible to use a baculovirus vector to delivery an NIS gene for non-invasive monitoring transplanted islets in vivo by the expression of the target gene. |
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