Sites responsible for infectivity and antigenicity on nervous necrosis virus (NNV) appear to be distinct

Hyun Jung Gye,Toyohiko Nishizawa

doi:10.1038/s41598-021-83078-3

Abstract

Nervous necrosis virus (NNV) is a pathogenic fish-virus belonging to the genus Betanodavirus (Nodaviridae). Surface protrusions on NNV particles play a crucial role in both antigenicity and infectivity. We exposed purified NNV particles to different physicochemical conditions to investigate the effects on antigenicity and infectivity, in order to reveal information regarding the conformational stability and spatial relationships of NNV neutralizing-antibody binding sites and cell receptor binding sites. Treatment with PBS at 37 °C, drastically reduced NNV antigenicity by 66–79% on day one, whereas its infectivity declined gradually from 107.6 to 105.8 TCID50/ml over 10 days. When NNV was treated with carbonate/bicarbonate buffers at different pHs, both antigenicity and infectivity of NNV declined due to higher pH. However, the rate of decline with respect to antigenicity was more moderate than for infectivity. NNV antigenicity declined 75–84% after treatment with 2.0 M urea, however, there was no reduction observed in infectivity. The antibodies used in antigenicity experiments have high NNV-neutralizing titers and recognize conformational epitopes on surface protrusions. The maintenance of NNV infectivity means that receptor binding sites are functionally preserved. Therefore, it seems highly likely that NNV neutralizing-antibody binding sites and receptor binding sites are independently located on surface protrusions.

Highlights

Nervous necrosis virus (NNV) is a pathogenic fish-virus belonging to the genus Betanodavirus (Nodaviridae)
In order to observe more detailed behaviors of surface protrusions to heat-denaturation, purified NNV particles immobilized on plate wells of enzyme-linked immunosorbent assay (ELISA) were treated with Dulbecco’s phosphate buffered saline (PBS) at 37 °C for 10 days
Treatments with PBS at 25 °C and deionized water (DIW) at 37 °C were performed as temperature and buffer controls (Fig. 1A,B)

Summary

Introduction

Nervous necrosis virus (NNV) is a pathogenic fish-virus belonging to the genus Betanodavirus (Nodaviridae). It has been reported that both infectivity and antigenicity of purified NNV declines after incubation at 45 °C due to denaturation of NNV surface p rotrusions[7].

Results

Conclusion