Immunogenicity in Rabbits of Virus-Like Particles from a Contemporary Rabbit Haemorrhagic Disease Virus Type 2 (GI.2/RHDV2/b) Isolated in The Netherlands

Qiuhong Miao; Ruibing Qi; Luut Veldkamp; Jooske Ijzer; Marja L Kik; Jie Zhu; Aoxing Tang; Dandan Dong; Yonghong Shi; Monique M van Oers; Guangqing Liu; Gorben P Pijlman

doi:10.3390/v11060553

Immunogenicity in Rabbits of Virus-Like Particles from a Contemporary Rabbit Haemorrhagic Disease Virus Type 2 (GI.2/RHDV2/b) Isolated in The Netherlands

Qiuhong Miao
, Ruibing Qi
, Luut Veldkamp
, Jooske Ijzer
, Marja L Kik
, Jie Zhu
, Aoxing Tang
, Dandan Dong
, Yonghong Shi
, Monique M van Oers
, Guangqing Liu
, Gorben P Pijlman

Laboratory of Virology, Wageningen University & Research, 6708 PB Wageningen, The Netherlands. [email protected].
Innovation Team of Small animal Infectious Disease, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Science, Shanghai 200241, China. [email protected].
Laboratory of Virology, Wageningen University & Research, 6708 PB Wageningen, The Netherlands. [email protected].
Innovation Team of Small animal Infectious Disease, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Science, Shanghai 200241, China. [email protected].
Innovation Team of Small animal Infectious Disease, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Science, Shanghai 200241, China. [email protected].
Innovation Team of Small animal Infectious Disease, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Science, Shanghai 200241, China. [email protected].
Innovation Team of Small animal Infectious Disease, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Science, Shanghai 200241, China. [email protected].
Laboratory of Virology, Wageningen University & Research, 6708 PB Wageningen, The Netherlands. [email protected].
Innovation Team of Small animal Infectious Disease, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Science, Shanghai 200241, China. [email protected].
Laboratory of Virology, Wageningen University & Research, 6708 PB Wageningen, The Netherlands. [email protected].

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Rabbit haemorrhagic disease virus (RHDV) type 2 (GI.2/RHDV2/b) is an emerging pathogen in wild rabbits and in domestic rabbits vaccinated against RHDV (GI.1). Here we report the genome sequence of a contemporary RHDV2 isolate from the Netherlands and investigate the immunogenicity of virus-like particles (VLPs) produced in insect cells. RHDV2 RNA was isolated from the liver of a naturally infected wild rabbit and the complete viral genome sequence was assembled from sequenced RT-PCR products. Phylogenetic analysis based on the VP60 capsid gene demonstrated that the RHDV2 NL2016 isolate clustered with other contemporary RHDV2 strains. The VP60 gene was cloned in a baculovirus expression vector to produce VLPs in Sf9 insect cells. Density-gradient purified RHDV2 VLPs were visualized by transmission electron microscopy as spherical particles of around 30 nm in diameter with a morphology resembling authentic RHDV. Immunization of rabbits with RHDV2 VLPs resulted in high production of serum antibodies against VP60, and the production of cytokines (IFN-γ and IL-4) was significantly elevated in the immunized rabbits compared to the control group. The results demonstrate that the recombinant RHDV2 VLPs are highly immunogenic and may find applications in serological detection assays and might be further developed as a vaccine candidate to protect domestic rabbits against RHDV2 infection.

Original language	English
Article number	doi:10.3390/v11060553
Number of pages	14
Journal	Viruses
Volume	11
Issue number	6
DOIs	https://doi.org/10.3390/v11060553
Publication status	Published - 14 Jun 2019

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

rabbit haemorrhagic disease virus 2 (GI
2/RHDV2/b)
Netherlands
VP60
baculovirus expression
virus-like particles
insect cells
immunogenicity

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10.3390/v11060553

viruses-11-00553-v2Final published version, 2.39 MB

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Miao, Q., Qi, R., Veldkamp, L., Ijzer, J., Kik, M. L., Zhu, J., Tang, A., Dong, D., Shi, Y., van Oers, M. M., Liu, G., & Pijlman, G. P. (2019). Immunogenicity in Rabbits of Virus-Like Particles from a Contemporary Rabbit Haemorrhagic Disease Virus Type 2 (GI.2/RHDV2/b) Isolated in The Netherlands. Viruses, 11(6), Article doi:10.3390/v11060553. https://doi.org/10.3390/v11060553

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title = "Immunogenicity in Rabbits of Virus-Like Particles from a Contemporary Rabbit Haemorrhagic Disease Virus Type 2 (GI.2/RHDV2/b) Isolated in The Netherlands",

abstract = "Rabbit haemorrhagic disease virus (RHDV) type 2 (GI.2/RHDV2/b) is an emerging pathogen in wild rabbits and in domestic rabbits vaccinated against RHDV (GI.1). Here we report the genome sequence of a contemporary RHDV2 isolate from the Netherlands and investigate the immunogenicity of virus-like particles (VLPs) produced in insect cells. RHDV2 RNA was isolated from the liver of a naturally infected wild rabbit and the complete viral genome sequence was assembled from sequenced RT-PCR products. Phylogenetic analysis based on the VP60 capsid gene demonstrated that the RHDV2 NL2016 isolate clustered with other contemporary RHDV2 strains. The VP60 gene was cloned in a baculovirus expression vector to produce VLPs in Sf9 insect cells. Density-gradient purified RHDV2 VLPs were visualized by transmission electron microscopy as spherical particles of around 30 nm in diameter with a morphology resembling authentic RHDV. Immunization of rabbits with RHDV2 VLPs resulted in high production of serum antibodies against VP60, and the production of cytokines (IFN-γ and IL-4) was significantly elevated in the immunized rabbits compared to the control group. The results demonstrate that the recombinant RHDV2 VLPs are highly immunogenic and may find applications in serological detection assays and might be further developed as a vaccine candidate to protect domestic rabbits against RHDV2 infection.",

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author = "Qiuhong Miao and Ruibing Qi and Luut Veldkamp and Jooske Ijzer and Kik, \{Marja L\} and Jie Zhu and Aoxing Tang and Dandan Dong and Yonghong Shi and \{van Oers\}, \{Monique M\} and Guangqing Liu and Pijlman, \{Gorben P\}",

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AU - Miao, Qiuhong

AU - Qi, Ruibing

AU - Veldkamp, Luut

AU - Ijzer, Jooske

AU - Kik, Marja L

AU - Zhu, Jie

AU - Tang, Aoxing

AU - Dong, Dandan

AU - Shi, Yonghong

AU - van Oers, Monique M

AU - Liu, Guangqing

AU - Pijlman, Gorben P

PY - 2019/6/14

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N2 - Rabbit haemorrhagic disease virus (RHDV) type 2 (GI.2/RHDV2/b) is an emerging pathogen in wild rabbits and in domestic rabbits vaccinated against RHDV (GI.1). Here we report the genome sequence of a contemporary RHDV2 isolate from the Netherlands and investigate the immunogenicity of virus-like particles (VLPs) produced in insect cells. RHDV2 RNA was isolated from the liver of a naturally infected wild rabbit and the complete viral genome sequence was assembled from sequenced RT-PCR products. Phylogenetic analysis based on the VP60 capsid gene demonstrated that the RHDV2 NL2016 isolate clustered with other contemporary RHDV2 strains. The VP60 gene was cloned in a baculovirus expression vector to produce VLPs in Sf9 insect cells. Density-gradient purified RHDV2 VLPs were visualized by transmission electron microscopy as spherical particles of around 30 nm in diameter with a morphology resembling authentic RHDV. Immunization of rabbits with RHDV2 VLPs resulted in high production of serum antibodies against VP60, and the production of cytokines (IFN-γ and IL-4) was significantly elevated in the immunized rabbits compared to the control group. The results demonstrate that the recombinant RHDV2 VLPs are highly immunogenic and may find applications in serological detection assays and might be further developed as a vaccine candidate to protect domestic rabbits against RHDV2 infection.

AB - Rabbit haemorrhagic disease virus (RHDV) type 2 (GI.2/RHDV2/b) is an emerging pathogen in wild rabbits and in domestic rabbits vaccinated against RHDV (GI.1). Here we report the genome sequence of a contemporary RHDV2 isolate from the Netherlands and investigate the immunogenicity of virus-like particles (VLPs) produced in insect cells. RHDV2 RNA was isolated from the liver of a naturally infected wild rabbit and the complete viral genome sequence was assembled from sequenced RT-PCR products. Phylogenetic analysis based on the VP60 capsid gene demonstrated that the RHDV2 NL2016 isolate clustered with other contemporary RHDV2 strains. The VP60 gene was cloned in a baculovirus expression vector to produce VLPs in Sf9 insect cells. Density-gradient purified RHDV2 VLPs were visualized by transmission electron microscopy as spherical particles of around 30 nm in diameter with a morphology resembling authentic RHDV. Immunization of rabbits with RHDV2 VLPs resulted in high production of serum antibodies against VP60, and the production of cytokines (IFN-γ and IL-4) was significantly elevated in the immunized rabbits compared to the control group. The results demonstrate that the recombinant RHDV2 VLPs are highly immunogenic and may find applications in serological detection assays and might be further developed as a vaccine candidate to protect domestic rabbits against RHDV2 infection.

KW - rabbit haemorrhagic disease virus 2 (GI

KW - 2/RHDV2/b)

KW - Netherlands

KW - VP60

KW - baculovirus expression

KW - virus-like particles

KW - insect cells

KW - immunogenicity

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SN - 1999-4915

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JO - Viruses

JF - Viruses

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M1 - doi:10.3390/v11060553

ER -

Immunogenicity in Rabbits of Virus-Like Particles from a Contemporary Rabbit Haemorrhagic Disease Virus Type 2 (GI.2/RHDV2/b) Isolated in The Netherlands

Abstract

UN SDGs

Keywords

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