Oral immunization with recombinant L. lactis expressing GCRV-II VP4 produces protection against grass carp reovirus infection

生物 免疫系统 免疫原性 病毒学 免疫 挑剔 抗原 微生物学 重组DNA 脾脏 草鱼 免疫学 渔业 基因 生物化学
作者
Huiliang Wu,Jiyuan Yin,Siming Li,Hao Wang,Peng Jiang,Pengfei Li,Zhaoyang Ding,Han Yan,Bo Chen,Li Wang,Qing Wang
出处
期刊:Fish & Shellfish Immunology [Elsevier]
卷期号:147: 109439-109439
标识
DOI:10.1016/j.fsi.2024.109439
摘要

The hemorrhagic disease causing by grass carp reovirus (GCRV) infection, is associated with major economic losses and significant impact on aquaculture worldwide. VP4 of GCRV is one of the major outer capsid proteins which can induce an immune response in the host. In this study, pNZ8148-VP4/L. lactis was constructed to express recombinant VP4 protein of GCRV, which was confirmed by the Western-Blot and enzyme-linked immunosorbent assay. Then we performed the oral immunization for rare minnow model and the challenge with GCRV-II. After oral administration, pNZ8148-VP4/L. lactis can continuously reside in the intestinal tract to achieve antigen presentation. The intestinal and spleen samples were collected at different time intervals after immunization, and the expression of immune-related genes was detected by real-time fluorescence quantitative PCR. The results showed that VP4 recombinant L. lactis could induce complete cellular and humoral immune responses in the intestinal mucosal system, and effectively regulate the immunological effect of the spleen. The immunogenicity and the protective efficacy of the oral vaccine was evaluated by determining IgM levels and viral challenge to vaccinated fish, a significant level (P < 0.01) of antigen-specific IgM with GCRV-II neutralizing activity was able to be detected, which provided a effective protection in the challenge experiment. These results indicated that an oral probiotic vaccine with VP4 expression can provide effective protection for grass carp against GCRV-II challenge, suggesting a promising vaccine strategy for fish.
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