Molecular cloning of the mRNA coding for the G protein of the viral haemorrhagic septicaemia (VHS) of salmonids

Thiry, Michel; Lecocq-Xhonneux, F.; Dheur, I.; Renard, A.; Kinkelin, P. de

doi:10.1016/0378-1135(90)90152-l

Cited by 22 publications

(7 citation statements)

References 15 publications

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“…The DNA sequence 24 corresponding to the glycoprotein G gene of VHSV 07.71, was used for the cloning experiments (Gene Bank accession number X59148).…”

Section: Construction Of Recombinant Glycoprotein G and Its Frg11 Fromentioning

confidence: 99%

In VitroNeutralization of Viral Hemorrhagic Septicemia Virus by Plasma from Immunized Zebrafish

Chinchilla

Gómez-Casado²,

Encinas³

et al. 2013

Zebrafish

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We studied humoral long-term adaptive viral neutralization responses in zebrafish (Danio rerio), an increasingly useful vertebrate model for viral diseases actually limited by the absence of standardized anti-zebrafish immunoglobulin M (IgM) antibodies. We established an alternative method, similar to those used in other fish, to achieve a first estimation of zebrafish anti-viral antibody-like responses. We used the viral hemorrhagic septicemia virus (VHSV) model because, although protection after this non-natural infection was demonstrated in cold-acclimatized zebrafish, little is known about their induced anti-VHSV antibody-like responses. Therefore, we first optimized a micro-neutralization method based on immunostaining VHSV-infected fish cell monolayers to detect zebrafish neutralizing activity in plasma samples in one day. We then used the method to measure the specific anti-VHSV neutralization in plasma obtained from individual zebrafish under various VHSV challenges or immunization protocols. The neutralizing activity was inhibited by protein A-sepharose and rabbit anti-zebrafish IgM antibodies, suggesting the implication of IgM zebrafish antibodies in such responses. To our knowledge, this is the first report to demonstrate detectable and significant VHSV neutralization titers in zebrafish surviving VHSV infections. This micro-method might be useful, not only for the follow-up of infection/vaccine development in the zebrafish/VHSV model in particular, but also for similar work involving other in vitro neutralizable zebrafish pathogens. This technique might also further the development of alternative ELISA assay methods to measure specific immunoglobulins in zebrafish.

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“…The DNA sequence 24 corresponding to the glycoprotein G gene of VHSV 07.71, was used for the cloning experiments (Gene Bank accession number X59148).…”

Section: Construction Of Recombinant Glycoprotein G and Its Frg11 Fromentioning

confidence: 99%

In VitroNeutralization of Viral Hemorrhagic Septicemia Virus by Plasma from Immunized Zebrafish

Chinchilla

Gómez-Casado²,

Encinas³

et al. 2013

Zebrafish

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“…Viral haemorrhagic septicaemia virus (VHSV) and infectious hematopoietic necrosis virus (IHNV) are fish rhabdoviruses classified in the genus Novirhabdovirus, responsible for systemic diseases costly for the salmonid farming industry. Six proteins are encoded by their non-segmented RNA genomes (Basurco & Benmansour, 1995;Benmansour et al, 1994;Bernard et al, 1990;Schütze et al, 1995Schütze et al, , 1999Thiry et al, 1990). Like in other rhabdoviruses, the glycoprotein (G) is the only protein exposed at the surface of VHSV and IHNV, and is responsible for the entry of the virus into the cell by receptor-mediated endocytosis.…”

Section: Introductionmentioning

confidence: 99%

The glycoprotein of a fish rhabdovirus profiles the virus-specific T-cell repertoire in rainbow trout

Boudinot

Bernard

Boubekeur

et al. 2004

Journal of General Virology

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T-cell responses to viruses are still poorly investigated in lower vertebrates. In rainbow trout, a specific clonal expansion of T cells in response to infection with viral haemorrhagic septicaemia virus (VHSV) was recently identified. Expanded T-cell clones expressed a unique 8 aa Vb4-Jb1 junction (SSGDSYSE) in different individuals, reminiscent of a typical public response. To get further insight into the nature of this response the modifications of the T-cell repertoire following immunization with plasmid expressing the VHSV external glycoprotein (G), which is the only protein involved in protective immunity, was analysed. After G-based DNA immunization, CDR3-length spectratypes were skewed for several Vb-Jb combinations, including Vb4-Jb1. In Vb4-Jb1, biases consisted of 6 and 8 aa junctions that were detected from day 52, and were still present 3 months after DNA immunization. Sequence analysis of the Vb4-Jb1 junctions showed that the 8 aa junction (SSGDSYSE) was clearly expanded, indicating that viral G protein was probably the target of the anti-VHSV public response. Additional 6 and 8 aa Vb4-Jb1 junctions were also expanded in G-DNA-vaccinated fish, showing that significant clonotypic diversity was selected in response to the plasmid-delivered G protein. This higher clonotypic diversity may be related to the demonstrated higher efficiency of G-based DNA vaccines over whole virus immunization. The use of infectious hematopietic necrosis virus (IHNV) recombinant viruses, expressing the VHSV G protein, further substantiated the VHSV G-protein specificity of the 8 aa Vb4-Jb1 response and designated the 6 aa Vb4-Jb1 response as potentially directed to a T-cell epitope common to VHSV and IHNV.

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“…It has been shown that the VHSV epitopes responsible for inducing protection are located on the surface glycoprotein (G), as is the case for other rhabdoviruses (Wiktor et al, 1973;Kelley et al, 1972;Engelking & Leong, 1989a, b), and that antibodies to the G protein are able to abolish virus 0001-2133 © 1994 SGM infectivity (de Kinkelin et al, 1984;Lorenzen et al, 1990). A cDNA copy of the mRNA coding for G protein of VHSV has been cloned (Thiry et al, 1990) and sequenced (Thiry et al, 1991). The VHSV G protein was also expressed in Escherichia coli, as a fusion protein with the bacterial enzyme TrpE but this protein failed to induce either consistent protection or the synthesis of neutralizing antibodies whatever the route of delivery (unpublished results).…”

Section: Introductionmentioning

confidence: 99%

A recombinant viral haemorrhagic septicaemia virus glycoprotein expressed in insect cells induces protective immunity in rainbow trout

Lecocq-Xhonneux

Thiry

Dheur

et al. 1994

Journal of General Virology

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Viral haemorrhagic septicaemia (VHS) is a fish rhabdovirus infection of world-wide importance. Control policies have been established but the disease still causes heavy losses in fish farming. The development of a recombinant subunit vaccine was initiated to produce a safe and effective vaccine to protect fish against VHS. The VHS virus (VHSV) glycoprotein, which induces neutralizing antibodies in rainbow trout, was chosen for expression in insect cells using a baculovirus vector. The Mr of the recombinant protein estimated by SDS-PAGE I was slightly lower than that of the native viral protein.The recombinant protein displayed different degrees of glycosylation and was recognized in ELISA by neutralizing antibodies. It was transported to the plasma membrane of insect cells where its ability to induce membrane fusion was preserved. The efficacy of the recombinant protein as a vaccine was compared with those of an inactivated and an attenuated vaccine. When injected intraperitoneally into rainbow trout, the baculovirus-encoded protein was shown (i) to induce the synthesis of VHSV-neutralizing antibodies and (ii) to confer protection against virus challenge. Immunization performed by immersion failed. This is the first report of a recombinant vaccine that protects fish against VHSV.

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Molecular cloning of the mRNA coding for the G protein of the viral haemorrhagic septicaemia (VHS) of salmonids

Cited by 22 publications

References 15 publications

In VitroNeutralization of Viral Hemorrhagic Septicemia Virus by Plasma from Immunized Zebrafish

In VitroNeutralization of Viral Hemorrhagic Septicemia Virus by Plasma from Immunized Zebrafish

The glycoprotein of a fish rhabdovirus profiles the virus-specific T-cell repertoire in rainbow trout

A recombinant viral haemorrhagic septicaemia virus glycoprotein expressed in insect cells induces protective immunity in rainbow trout

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