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Vet. Sci. 2018, 5(2), 57; https://doi.org/10.3390/vetsci5020057

Efficient Transduction and Expansion of Ovine Macrophages for Gene Therapy Implementations

1
Laboratory of Microbiology and Infectious Diseases, Faculty of Veterinary Medicine, Aristotle University of Thessaloniki, Thessaloniki 54124, Greece
2
Laboratory of Biology, School of Medicine, National and Kapodistrian University of Athens, Athens 11527, Greece
*
Authors to whom correspondence should be addressed.
Received: 6 May 2018 / Revised: 5 June 2018 / Accepted: 14 June 2018 / Published: 18 June 2018
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Abstract

A number of bacteria provoking zoonotic diseases present intracellular survival and a host cell tropism limited to the monocyte/macrophage lineage. Thus, infection is rendered difficult to eradicate, causing chronic inflammatory reactions to the host and widespread prevalence. Although self-inactivating lentiviral vectors have been successfully tested in the clinic against virally-induced human infectious diseases, little is known about the transduction susceptibility of ruminant animal phagocytes that play a critical role in the outbreak of zoonotic diseases such as brucellosis. In view of the development of a lentiviral vector-based platform targeting and inactivating specific genetic features of intracellular bacteria, we have tested the transducibility of ovine macrophages in terms of transgene expression and vector copy number (VCN). We show that ovine macrophages are relatively resistant to transduction even at a high multiplicity of infection with a conventional lentiviral vector expressing the green fluorescence protein and that addition of transduction enhancers, such as polybrene, increases transgene expression even after a one-week culture of the transduced cells in vitro. Overall, we demonstrate that ovine macrophages may be efficiently expanded and transduced in culture, thus providing the benchmark for gene therapy applications for zoonotic diseases. View Full-Text
Keywords: zoonotic diseases; macrophages; ruminant animals; lentiviral vectors; transduction; gene therapy zoonotic diseases; macrophages; ruminant animals; lentiviral vectors; transduction; gene therapy
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Karponi, G.; Kritas, S.; Petridou, E.; Papanikolaou, E. Efficient Transduction and Expansion of Ovine Macrophages for Gene Therapy Implementations. Vet. Sci. 2018, 5, 57.

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