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Viruses 2018, 10(10), 574; https://doi.org/10.3390/v10100574

Improved Baculovirus Vectors for Transduction and Gene Expression in Human Pancreatic Islet Cells

1
Department of Biological and Medical Sciences, Oxford Brookes University, Oxford OX3 0BP, UK
2
Oxford Expression Technologies Ltd., Bioinnovation Hub, Gipsy Lane Campus, Oxford OX3 0BP, UK
3
Department of Biotechnology, College of Sciences, Baghdad University, Baghdad 10071, Iraq
4
Centre for Molecular and Cell-Based Therapeutics SA de CV, Mexico City 15820, Mexico
5
Nuffield Department of Surgical Sciences, University of Oxford, Oxford OX3 9DU, UK
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Received: 5 October 2018 / Accepted: 18 October 2018 / Published: 20 October 2018
(This article belongs to the Special Issue Baculovirus Advances and Applications)
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Abstract

Pancreatic islet transplantation is a promising treatment for type 1 diabetes mellitus offering improved glycaemic control by restoring insulin production. Improved human pancreatic islet isolation has led to higher islet transplantation success. However, as many as 50% of islets are lost after transplantation due to immune responses and cellular injury, gene therapy presents a novel strategy to protect pancreatic islets for improved survival post-transplantation. To date, most of the vectors used in clinical trials and gene therapy studies have been derived from mammalian viruses such as adeno-associated or retrovirus. However, baculovirus BacMam vectors provide an attractive and safe alternative. Here, a novel BacMam was constructed containing a frameshift mutation within fp25, which results in virus stocks with higher infectious titres. This improved in vitro transduction when compared to control BacMams. Additionally, incorporating a truncated vesicular stomatitis virus G protein increased transduction efficacy and production of EGFP and BCL2 in human kidney (HK-2) and pancreatic islet β cells (EndoC βH3). Lastly, we have shown that our optimized BacMam vector can deliver and express egfp in intact pancreatic islet cells from human cadaveric donors. These results confirm that BacMam vectors are a viable choice for providing delivery of transgenes to pancreatic islet cells. View Full-Text
Keywords: BacMam; baculovirus; gene therapy; high-titre virus; human pancreatic islet cells BacMam; baculovirus; gene therapy; high-titre virus; human pancreatic islet cells
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Graves, L.P.; Aksular, M.; Alakeely, R.A.; Ruiz Buck, D.; Chambers, A.C.; Murguia-Meca, F.; Plata-Muñoz, J.-J.; Hughes, S.; Johnson, P.R.V.; Possee, R.D.; King, L.A. Improved Baculovirus Vectors for Transduction and Gene Expression in Human Pancreatic Islet Cells. Viruses 2018, 10, 574.

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