| Publication number |
CLPUB00590 |
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| Authors |
Nunez-Ortiz N., Buonocore F., Stocchi V., Picchietti S., Randelli E., Toffan A., Pascoli F., Scapigliati G. |
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| Title |
The encephalopathy and retinopathy virus of European sea bass dicentrarchus labrax: strategies for its detection and immunisation of juveniles. |
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| Citation |
Bull. Eur. Assoc. Fish Pathol. 36:155-163(2016) |
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| Web pages |
https://eafp.org/download/2016-volume36/issue_4/36-4-155-nunez.pdf |
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| Abstract |
Viral encephalopathy and retinopathy virus (VERv) represents a major
threat for many wild and farmed fish species among which the European sea
bass is preferentially infected during young life stages. We are currently
investigating the possibilities of immunising young sea bass against VERv
by means of mucosal (immersion) vaccination employing inactivated VERv.
VERv-free sea bass of 2-10 grams were immunised by immersion in virus
solution for various times and then sampled after 24, 48, 72 hours, and
after 30 days. The results of this round of immunisations showed that: i)
no serum antigen-specific IgM can be detected by ELISA; no in vitro VERv-
induced gill leukocyte proliferation can be induced; iii) a modulation in
transcription levels of genes coding for IFN, Mx, ISG-12, IgT was observed
after 24 hours; iv) immunohistochemistry showed the presence of VERv
antigens. These results show that a single immersion vaccination without
boosting does not induce systemic IgM-based immunity in sea bass. A
control experiment where VERv was administered intraperitoneally showed
the presence of serum antigen-specific IgM. The sea bass cell line DLEC
has been transfected with a plasmid encoding VERv capsid protein, and
transfected cells have been used to immunise juveniles intraperitoneally.
After 30 days sera and leukocytes were collected, and results showed that
sera of fish were positive in ELISA against adsorbed VERv and that
leukocytes from spleen and head kidney proliferated in vitro in response
to VERv, suggesting interesting possibilities for this system to be
employed to investigate antiviral immune responses. Finally, by employing
a rabbit antiserum against VERv (pAb 283) and a mouse monoclonal against
VERv capsid protein (mAb 4C3) we have developed a Capture ELISA system to
detect and quantitate the presence of VERv in solutions and biological
fluids. Overall, our results represent a first comprehensive approach to
detect and investigate the effects of VERv immunisation of sea bass.
|
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| Cell lines |
CVCL_6F79; DLEC CVCL_A1QR; DLEC-NNV |
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