Novel vectors for simultaneous high-level dual protein expression in vertebrate and insect cells by recombinant baculoviruses

Keil, Günther M.; Klopfleisch, Constanze; Giesow, Katrin; Blohm, Ulrike

doi:10.1016/j.jviromet.2009.05.001

Cited by 14 publications

(5 citation statements)

References 22 publications

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“…To induce the differentiation to myotubes, the medium of a 100% confluent L6 myoblast culture was replaced with medium containing only 1% FBS. 34 To transduce cells with the recombinant baculovirus, we followed the procedure described in Keil et al 57 Briefly, medium in six-well plates with confluent L6 or MA104 cells was replaced with 1.5 ml Dulbecco's modified Eagle's medium without serum or antibiotics, and the baculovirus particles were added (10 or 100 p.f.u. per cell).…”

Section: Pcr Analysis In Genomic Dna From Baculovirus Particlesmentioning

confidence: 99%

Delivery of glutamine synthetase gene by baculovirus vectors: a proof of concept for the treatment of acute hyperammonemia

et al. 2014

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Hyperammonemia, a condition present in patients with urea cycle disorders (UCDs) or liver diseases, can cause neuropsychiatric complications, which in the worst cases result in brain damage, coma or death. Diverse treatments exist for the treatment of hyperammonemia, but they have limited efficacy, adverse effects and elevated cost. Gene therapy is a promising alternative that is explored here. A baculovirus, termed Bac-GS, containing the glutamine synthetase (GS) gene was constructed for the in vitro and in vivo treatment of hyperammonemia. Transduction of MA104 epithelial or L6 myoblast/myotubes cells with Bac-GS resulted in a high expression of the GS gene, an increase in GS concentration, and a reduction of almost half of exogenously added ammonia. When Bac-GS was tested in an acute hyperammonemia rat model by intramuscularly injecting the rear legs, the concentration of ammonia in blood decreased 351 μM, in comparison with controls. A high GS concentration was detected in gastrocnemius muscles from the rats transduced with Bac-GS. These results show that gene delivery for overexpressing GS in muscle tissue is a promising alternative for the treatment of hyperammonemia in patients with acute or chronic liver diseases and hepatic encephalopathy or UCD.

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Section: Pcr Analysis In Genomic Dna From Baculovirus Particlesmentioning

confidence: 99%

Delivery of glutamine synthetase gene by baculovirus vectors: a proof of concept for the treatment of acute hyperammonemia

et al. 2014

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“…Over the past few years, a wealth of literature has addressed the problems in baculoviral vector production Carinhas et al, 2009Carinhas et al, , 2010Dojima et al, 2010;Tsai et al, 2007), quantification (Chan et al, 2006;Ferris et al, 2011;Kärkkäinen et al, 2009;Lo et al, 2011;Lo and Chao, 2004;Roldao et al, 2009), purification (Chen et al, 2009a;Kaikkonen et al, 2008;Transfiguracion et al, 2007;Vicente et al, 2009Vicente et al, , 2010aVicente et al, , 2010bWu et al, 2007) and quality assurance (Ihalainen et al, 2010;Jorio et al, 2006). New baculoviral vectors taking advantage of hybrid promoters (Gao et al, 2007;Keil et al, 2009;Lackner et al, 2008;Pan et al, 2009) and new regulatory elements (Du et al, 2010;Mahonen et al, 2007) are also being developed and evaluated for their potential applications. Additionally, the transduction conditions (Keil et al, 2009;Pan et al, 2009), supplements (Guo et al, 2010;Yin et al, 2010) and parameters (Lee et al, 2007a(Lee et al, , 2007bShen et al, 2007Shen et al, , 2008 dictating the transduction efficiencies have been evaluated.…”

Section: Discussionmentioning

confidence: 99%

“…New baculoviral vectors taking advantage of hybrid promoters (Gao et al, 2007;Keil et al, 2009;Lackner et al, 2008;Pan et al, 2009) and new regulatory elements (Du et al, 2010;Mahonen et al, 2007) are also being developed and evaluated for their potential applications. Additionally, the transduction conditions (Keil et al, 2009;Pan et al, 2009), supplements (Guo et al, 2010;Yin et al, 2010) and parameters (Lee et al, 2007a(Lee et al, , 2007bShen et al, 2007Shen et al, , 2008 dictating the transduction efficiencies have been evaluated. These technological progresses undoubtedly will facilitate the production of baculoviral vectors in compliance of cGMP regulations, and advance baculovirus from a research tool to clinical applications.…”

Section: Discussionmentioning

confidence: 99%

Baculovirus as a gene delivery vector: Recent understandings of molecular alterations in transduced cells and latest applications

Chen

Lin

Chen

et al. 2011

Biotechnology Advances

128

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Baculovirus infects insects in nature and is non-pathogenic to humans, but can transduce a broad range of mammalian and avian cells. Thanks to the biosafety, large cloning capacity, low cytotoxicity and non-replication nature in the transduced cells as well as the ease of manipulation and production, baculovirus has gained explosive popularity as a gene delivery vector for a wide variety of applications. This article extensively reviews the recent understandings of the molecular mechanisms pertinent to baculovirus entry and cellular responses, and covers the latest advances in the vector improvements and applications, with special emphasis on antiviral therapy, cancer therapy, regenerative medicine and vaccine.

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“…In vAc-HA-DUAL, the expression of the egfp reporter and HA genes was driven by two hybrid promoters, composed either of the baculovirus p10 and mammalian CMV promoters, or by the baculovirus polyhedrin and CMV promoters, respectively (Figure 1). Although the transcription mediated by these hybrid promoters requires further detailed characterization, they both appeared highly effective in insect and mammalian cells [22,23] .…”

Section: Discussionmentioning

confidence: 99%

Characterization of the immune responses elicited by baculovirus-based vector vaccines against influenza virus hemagglutinin

Yin

Zhang

et al. 2012

Acta Pharmacol Sin

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Aim:To compare the specific immune responses elicited by different baculovirus vectors in immunized mice. Methods: We constructed and characterized two recombinant baculoviruses carrying the expression cassette for the H5N1 influenza virus hemagglutinin (HA) gene driven by either an insect cell promoter (vAc-HA) or a dual-promoter active both in insect and mammalian cells (vAc-HA-DUAL). Virus without the HA gene (vAc-EGFP) was used as a control. These viruses were used to immunize mice subcutaneously and intraperitoneally. The production of total and specific antibodies was determined by ELISA and competitive ELISA. Cytokine production by the spleen cells of immunized mice was studied using the ELISPOT assay. Results: Both the vAc-HA and vAc-HA-DUAL vectors expressed HA proteins in insect Sf9 cells, and HA antigen was present in progeny virions. The vAc-HA-DUAL vector also mediated HA expression in virus-transduced mammalian cell lines (BHK and A547). Both vAc-HA and vAc-HA-DUAL exhibited higher transduction efficiencies than vAc-EGFP in mammalian cells, as shown by the expression of the reporter gene egfp. Additionally, both vAc-HA and vAc-HA-DUAL induced high levels of HA-specific antibody production in immunized mice; vAc-HA-DUAL was more efficient in inducing IFN-γ and IL-2 upon stimulation with specific antigen, whereas vAc-HA was more efficient in inducing IL-4 and IL-6. Conclusion: Baculovirus vectors elicited efficient, specific immune responses in immunized mice. The vector displaying the HA antigen on the virion surface (vAc-HA) elicited a Th2-biased immune response, whereas the vector displaying HA and mediating HA expression in the cell (vAc-HA-DUAL) elicited a Th1-biased immune response.

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Novel vectors for simultaneous high-level dual protein expression in vertebrate and insect cells by recombinant baculoviruses

Cited by 14 publications

References 22 publications

Delivery of glutamine synthetase gene by baculovirus vectors: a proof of concept for the treatment of acute hyperammonemia

Delivery of glutamine synthetase gene by baculovirus vectors: a proof of concept for the treatment of acute hyperammonemia

Baculovirus as a gene delivery vector: Recent understandings of molecular alterations in transduced cells and latest applications

Characterization of the immune responses elicited by baculovirus-based vector vaccines against influenza virus hemagglutinin

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