Immune response against grouper nervous necrosis virus by vaccination of virus-like particles.

The grouper is a high-value fish in the seafood market. Grouper nervous necrosis virus (GNNV) causes mass mortality, near 100% in larvae and juveniles, which has great economic impact on the aquaculture of the marine fish. Since vaccination is one of the best methods against viral diseases, grouper Epinephelus lanceolatus was injected with virus-like particles (VLPs) of GNNV at different dosages and injection frequencies. The anti-sera of vaccinated fish were analyzed with antigen-capture ELISA to quantify immunization titer. The antibody titers in the vaccinated fish increased remarkably within 4 weeks, during which time the antibody was definitely capable to neutralize the native virus. With one shot of 10-250 microg VLPs, the stimulated antibody titer reached a steady saturation level in 1 month, among which the titers by one shot of 100 and 250 microg VLPs were 13% higher than by 10 microg. Two shots of 10 and 100 microg VLPs increased to maximum titer, which was 29% higher than one shot, whereas two shots of 250 microg VLPs and four shots of 100 microg VLPs dramatically downgraded the titers by -23% and -44%, respectively. These results imply that the overdose effects occurred in total dosages higher than 200 microg VLPs. The experiments of VLP vaccine with adjuvant revealed that the adjuvant is not required for increasing the efficacy of the VLP vaccine. Immunization with the VLPs can also stimulate fish to produce high antibody titer for more than 5 months, which can be correlated to long-term protection. When VLPs are used as vaccine agent, a dosage at 1 microg/g of fish body weight is enough to stimulate a full-scale immune response.