GCRV 096 VP6 protein and its impacts on GCRV replication with different genotypes in CIK cells

Yan, Xin; Xiong, Lingfang; Li, Jie; Ya, Wang; Wu, Zaohe; Jian, Jichang; Ding, Yu

doi:10.1016/j.aaf.2018.07.005

Cited by 5 publications

(7 citation statements)

References 35 publications

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“…3 and 4 indicate that the replication of GCRV 096 was inhibited in the samples incubated with antiserum against the GCRV 096 NS38 protein. Moreover, we found that the expressed soluble NS38 protein elicited a higher antibody titre than the GCRV 096 VP6 protein did (Yan et al, 2018). Recent researches have shown viral non-structural proteins as promising vaccine candidates and major targets for antiviral therapy (Hurtado-Melgoza et al, 2016;Chu et al, 2017;Kitagawa et al, 2017;Reyes-Sandoval and Ludert, 2019).…”

Section: Replication Of Gcrvs In Cik Cellsmentioning

confidence: 80%

“…ELISA plates with the expressed NS38 protein were coated with the antiserum raised against GCRV 096 NS38 protein for indirect ELISA assay. The experimental results indicated that the titre of the antiserum was 1:38400, higher than that of the GCRV 096 VP6 protein (Yan et al, 2018). Western blot showed that the antiserum could specifically bind the NS38 protein (Fig.…”

Section: Indirect Plate Elisa and Western Blot Analyses Using Antiserum Raised Against The Gcrv 096 Ns38 Proteinmentioning

confidence: 91%

“…Normal serum (prior to immunisation) was collected from the Zealand white rabbits, as the negative control. Equal amounts of the purified NS38 protein (500 µg) and Freund's complete adjuvant (Sigma, America) were mixed and emulsified completely and antiserum against the GCRV 096 NS38 protein was raised as previously described (Yan et al, 2018). The prepared antiserum was separated and stored at -20°C.…”

Section: Preparation Of Antiserum Against the Gcrv 096 Ns38 Proteinmentioning

confidence: 99%

“…The infected CIK cells and media were centrifuged at 35000 g, at 4°C for 2.5 h. Then, proteins were extracted from the precipitate for western blot (Yan et al, 2018). Antibodies used were mouse monoclonal anti-β-Tubulin antibody (1:1000; Invitrogen), mouse monoclonal anti-NS38 antibody (prepared in Vipotion, China) and goat anti-mouse antibody (1:3000; Takara, China).…”

Section: Qrt-pcr Amplification Statistical Analysis and Western Blotmentioning

confidence: 99%

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NS38 protein of GCRV 096: Functional analysis and effect on grass carp reovirus replication

Yan¹,

Xiong²,

Li³

et al. 2021

Indian Journal of Fisheries

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Grass carp reovirus (GCRV) seriously affects the grass carp farming, as the causative agent of grass carp hemorrhagic disease. In this study, we found the GCRV 096 NS38 protein contained one conservative PXXP motif similar to σNS of avian reovirus, with very low similarity rates with its homologous GCRV proteins of different genotypes. Furthermore, the soluble GCRV 096 NS38 protein was expressed in Escherichia coli, antiserum was raised and characterised. Addition of purified NS38 protein to culture media of Ctenopharyngodon idella kidney (CIK) cells clearly inhibited replication of GCRV 096 (genotype I); however, replication of GCRV GD108 (genotype Ш) was not significantly affected in CIK cells under the same conditions. It was not possible to induce cross-protection among GCRVs of different genotypes. GCRV multi-genotype should be considered and the NS38 protein was a promising candidate for developing therapies against grass carp hemorrhagic disease. Additionally, bioinformatics analysis suggested that the NS38 protein may be involved in viral genome replication.

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Section: Replication Of Gcrvs In Cik Cellsmentioning

confidence: 80%

Section: Indirect Plate Elisa and Western Blot Analyses Using Antiserum Raised Against The Gcrv 096 Ns38 Proteinmentioning

confidence: 91%

Section: Preparation Of Antiserum Against the Gcrv 096 Ns38 Proteinmentioning

confidence: 99%

Section: Qrt-pcr Amplification Statistical Analysis and Western Blotmentioning

confidence: 99%

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NS38 protein of GCRV 096: Functional analysis and effect on grass carp reovirus replication

Yan¹,

Xiong²,

Li³

et al. 2021

Indian Journal of Fisheries

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“…As with all reoviruses, GCRV II is organized as two concentric icosahedral layers: an inner core and an outer capsid layer ( 16 ). VP6 is one of the primary structural protein components of the GCRV II mature particle and is encoded by segment 9 and forms part of the viral core and plays an important role in replication ( 17 ). In particular, VP6 functions as a mediator that bridges the inner core and outer capsid and is directly inserted into position co-translationally without the involvement of ATP or chaperones.…”

Section: Introductionmentioning

confidence: 99%

Recombinant Lactococcus lactis Expressing Grass Carp Reovirus VP6 Induces Mucosal Immunity Against Grass Carp Reovirus Infection

Wang

et al. 2022

Front. Immunol.

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Grass carp haemorrhagic disease caused by grass carp reovirus II is a serious disease of the aquaculture industry and vaccination is the only effective method of GCRV protection. In this study, Lactococcus lactis was used as oral vaccine delivery to express the GCRV II VP6 protein. We evaluated the protective efficacy of the live vaccine strain to induce mucosal immune protection. After oral administration, the recombinant strains remained in the hindgut for antigen presentation and increased the survival rate 46.7% and the relative percent survival 42.9%, respectively versus control vaccination. Though L. lactis alone can induce the inflammatory response by stimulating the mucosal immune system, the recombinant L. lactis expressing VP6 greatly enhanced nonspecific immune responses via expression of immune related genes of the fish. Furthermore, both systemic and mucosal immunity was elicited following oral immunization with the recombinant strain and this strain also elicited an inflammatory response and cellular immunity to enhance the protective effect. L. lactis can therefore be utilized as a mucosal immune vector to trigger high levels of immune protection in fish at both the systemic and mucosal levels. L. lactis is a promising candidate for oral vaccine delivery.

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Establishment of a rare minnow (Gobiocypris rarus) disease model for grass carp reovirus genotype II

Chen

Chang

et al. 2021

Aquaculture

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GCRV 096 VP6 protein and its impacts on GCRV replication with different genotypes in CIK cells

Cited by 5 publications

References 35 publications

NS38 protein of GCRV 096: Functional analysis and effect on grass carp reovirus replication

NS38 protein of GCRV 096: Functional analysis and effect on grass carp reovirus replication

Recombinant Lactococcus lactis Expressing Grass Carp Reovirus VP6 Induces Mucosal Immunity Against Grass Carp Reovirus Infection

Establishment of a rare minnow (Gobiocypris rarus) disease model for grass carp reovirus genotype II

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