Virus-specific protein synthesis in cells infected by infectious pancreatic necrosis virus

An investigation of virus-specific protein maturation in infectious pancreatic necrosis virus (IPNV) infected Chinook salmon embryo cells (CHSE-214) was undertaken. The precursor protein (pVP2-1) of the major mature capsid protein (VP2) was processed sequentially from pVP2-1 to pVP2-2 and VP2. Experiments using serine proteinase inhibitors showed that the maturation of the VP2 was blocked in the pVP2-1 post-translational cleavage steps. A protinin, a potent proteinase inhibitor, at 800 μg ml(-1) blocked pVP2-2 to VP2 and the cleavage of VP4 (28 kDa) to VP4-1 (25 kDa). Therefore, our data showed that the maturation of the capsid protein (VP2) and cleavage of VP4 (NS proteinase) can be blocked by serine proteinase inhibitors.

Section: Discussionmentioning

confidence: 99%

Involvement of serine proteinase in infectious pancreatic necrosis virus capsid protein maturation and NS proteinase cleavage in CHSE‐214 cells

Hong

Chang

et al. 1998

“…One possible explanation is the difference in the quantity of viral mRNAs in infeeted cells. The number of struetural proteins in the virion is higher than the number of RNA-polymerase molecules (Dobos 1977). which suggests that the synthesis of mRNAs from segment A may be higher than from segment B.…”

Section: Discussionmentioning

confidence: 96%

Use of cloned cDNA probes for diagnosis of infectious pancreatic necrosis virus infections

Dopazo

Hetrick

Samal

1994

A dot-blot hybridization test has been developed lor the detection of infectious pancreatic necrosis virus (IPNV) in infected fish. For this purpose, cloning of the dsRNA of the West Buxton strain of IPNV was carried out. Two cDNA clones (WB and A4) were characterized for use as diagnostic probes and corresponded lo IPNV genome segments A and B, respectively. Clone WBl, with an insert of 812 base pairs, showed an 87 and 77% nuclcotidc sequence homology with the corresponding sequences of Jasper and Nl strains, respectively. Clone A4, with an insert size of 596bp, presented a nucleotide sequence homology of 90 and 80% with the corresponding sequences of the Jasper and Sp strains, respectively. Both probes were able to detect 15 ng of purified dsRNA, and were highly efficient in detecting the RNA of American IPNV strains. However, the A4 probe was less effective than WBl in hybridizing to RNA from European and Spanish strains of IPNV. Both probes detected IPNV RNA in ceils 4-8h post-infection with the homologous West Buxton strain, 8-12h post-infection with other American strains and 24h post infection with the European strains of IPNV. The method was less sensitive in detecting IPNV RNA directly in infected fish tissues. However, the present authors obtained a 100% effectiveness to detect viral RNA in cells inoculated with fish tissues confirmed by conventional diagnostic methods as being infected with IPNV. Therefore, the hybridization test is appropriate if combined with conventional diagnostic procedures, e.g. applying the dot blot hybridization test on tissue cultures 12-24 h after inoculation with infected fish tissue homoccnatcs.

“…The We report here the production of 31 MAbs against YAV which reacted with virus-infected cells by an immunofluorescence test. Viruses belonging to the family Birnaviridae possess a bisegmented dsRNA genome and four structural polypeptides, designated VP], VP2, VP3 and NS (VP4), of which VP2 is the most abundant (Dobos 1977(Dobos , 1991. In our trials, all but one of the virus-speciftc MAbs obtained were reactive with VP2 or VP3 polypeptides, which constitute the major capsid polypeptides encoded in the larger of the two genome segments (Duncan, Nagy, Krell & Dobos 1987).…”

Section: Discussionmentioning

confidence: 99%

Production of monoclonal antibodies against yellowtail ascites virus

Nakajima

Sorimachi

1996

A total of 31 antibody-secreting hybridoma cells against the yellowtail ascites virus (YAV) were established. These monoclonal antibodies (MAbs) reacted with the cytoplasm of YAV-infecred CHSE-214 cells, but not with uninfected CHSE-214 cells in an immunofluorescence test. Using these MAbs, two classes of polypeptides (\rP2 and VP3) were characterized by immunoprecipiration followed by SDS-PAGE, although one MAb did not react with either polypeptide. Eifteen out of the 17 MAbs that were reactive with VP2 polypeptides neutralized virus infeetivity, but all 13 MAbs that were reactive with VP3 did not neutralize infeetivity. In the immunofluorescence test, 29 out of the 31 MAbs ohtained showed the same reaction pattern to 12 YAV isolates from yellowtail, Seriola quinqueradiata, goldstriped amberjack, Seriola aureovittata, and threeline grunt, Parapristipoma trilineatum, from different geographical regions. The remaining two MAbs showed slightly different reaction patterns to the YAV isolates. The reaction patterns of the MAbs to the VR-299, Sp and Ab strains of IPNV were also investigated. Eourteen MAbs reacted to all three IPNV strains. The other 17 MAbs showed a negative reaction with at least one strain.