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Crude extracts of recombinant baculovirus expressing rabbit hemorrhagic disease virus 2 VLPs from both insect and rabbit cells protect rabbits from rabbit hemorrhagic disease caused by RHDV2

  • Claudia Müller
  • Reiner Ulrich
  • Kati Franzke
  • Marcus Müller
  • Bernd Köllner
Original Article

Abstract

Vaccines against viral pathogens are often composed of recombinant proteins expressed in different systems. Such proteins expressed by recombinant baculoviruses have been proven to be effective for vaccination. Especially, after codon usage optimization high amounts of recombinant viral proteins can be obtained which can assemble to virus like particles (VLPs) spontaneously. In this study we compared two different codon usages of RHDV2-VP1 to improve the expression of recombinant VP1 of RHDV2 by recombinant baculoviruses after infection of insect SF9 cells or transduction of mammalian RK13 cells in order to gain high protein yields. Also the influence on the auto-assembly of RHDV2-VP1 to VLPs was investigated. Finally, the immunogenic potential of such recombinant vaccines against RHDV2 to induce a protective immune response in rabbits against RHDV2 should be characterized. There was no influence of different codon usages on RHDV2-VP1 gene expression in the respective cell lines detected. However, in insect cell line SF9 higher rates of recombinant VP1 were measured in comparison to the transduction of mammalian cells RK13. Auto-assembly of RHDV2-VP1 to VLPs was observed in both cell systems by electron microscopy. Finally, both RHDV-VP1 VLPs derived from mammalian and insect cells were able to induce a protective humoral immune response in rabbits against RHDV2.

Notes

Acknowledgement

The author’s thank Dr. Günther Keil and Dr. Horst Schirrmeier for their methodological help. The excellent technical assistance of Katrin Giesow, Bianka Hillmann, Petra Meyer, Mandy Jörn and Sabine Weber is gratefully acknowledged.

Funding

This study was funded by IDT Biologika (Riems), Greifswald-Insel Riems, Germany for C.M.

Compliance with ethical standards

Conflict of interest

The authors Claudia Müller, Reiner Ulrich, Kati Franzke, Marcus Müller, and Bernd Köllner declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. Prior to the start of all trials the approval from the Federal state Ethical Committee for Animal Experimentation (LALLF-7221.3-1-025/15) was given and was performed following the acquirements of the EU directive 2010/63 and the EG recommendation 2007/526/.

Human and animal rights

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

705_2018_4032_MOESM1_ESM.docx (23 kb)
Suppl. 1 Tab. Primers and probes used for real time RT-PCR. Suppl. 2. Gene sequences of the artificial genes of RHDV2-VP1 with the different codon usage of “BHV-1” or “AcMNPV” (DOCX 23 kb)

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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Institute of Molecular Virology and Cell BiologyFriedrich-Loeffler-InstituteGreifswald-Insel RiemsGermany
  2. 2.Department of Experimental Animal Facilities and Biorisk ManagementFriedrich-Loeffler-InstituteGreifswald-Insel RiemsGermany
  3. 3.Institute of InfectologyFriedrich-Loeffler-InstituteGreifswald-Insel RiemsGermany
  4. 4.Institute of ImmunologyFriedrich-Loeffler-InstituteGreifswald-Insel RiemsGermany
  5. 5.IDT Biologika (Riems)Greifswald-Insel RiemsGermany

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