Müller Claudia, Ulrich Reiner, Franzke Kati, Müller Marcus, Köllner Bernd
Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institute, Greifswald-Insel Riems, Germany.
Department of Experimental Animal Facilities and Biorisk Management, Friedrich-Loeffler-Institute, Greifswald-Insel Riems, Germany.
Arch Virol. 2019 Jan;164(1):137-148. doi: 10.1007/s00705-018-4032-2. Epub 2018 Oct 5.
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.
针对病毒病原体的疫苗通常由在不同系统中表达的重组蛋白组成。由重组杆状病毒表达的此类蛋白已被证明对疫苗接种有效。特别是,在密码子使用优化后,可以获得大量的重组病毒蛋白,这些蛋白可以自发组装成病毒样颗粒(VLP)。在本研究中,我们比较了兔出血症病毒2(RHDV2)-VP1的两种不同密码子使用情况,以提高重组杆状病毒在感染昆虫SF9细胞或转导哺乳动物RK13细胞后RHDV2重组VP1的表达,从而获得高蛋白产量。还研究了其对RHDV2-VP1自组装成VLP的影响。最后,应表征此类抗RHDV2重组疫苗在兔体内诱导针对RHDV2的保护性免疫反应的免疫原性潜力。在各自的细胞系中未检测到不同密码子使用对RHDV2-VP1基因表达的影响。然而,与哺乳动物细胞RK13的转导相比,在昆虫细胞系SF9中测得的重组VP1率更高。通过电子显微镜在两种细胞系统中均观察到RHDV2-VP1自组装成VLP。最后,源自哺乳动物和昆虫细胞的两种RHDV-VP1 VLP均能够在兔体内诱导针对RHDV2的保护性体液免疫反应。