Abstract

White spot syndrome virus (WSSV) is one of the most devastating pathogens in penaeid shrimp and can cause massive damage in shrimp aquaculture industries. Previously, the WSSV structural protein VP15 was identified as an antigenic reagent against WSSV infections. In this study, we truncated this protein into VP15(1–25), VP15(26–57), VP15(58–80), and VP15(1–25,58–80). The purified proteins from the E. coli expression system were assayed as potential protective agents in Kuruma shrimp (Marsupenaeus japonicus) using the prime-and-boost strategy. Among the four truncated constructs, VP15(26–57) provided a significant improvement in the shrimp survival rate after 20 days of viral infection. Subsequently, four peptides (KR11, SR11, SK10, and KK13) from VP15(26–57) were synthesized and applied in an in vivo assay. Our results showed that SR11 could significantly enhance the shrimp survival rate, as determined from the accumulated survival rate. Moreover, a multiligand binding protein with a role in the host immune response and a possible VP15-binding partner, MjgC1qR, from the host M. japonicus were employed to test its binding with the VP15 protein. GST pull-down assays revealed that MjgC1qR binds with VP15, VP15(26–57), and SR11. Taken together, we conclude that SR11 is a determinant antigenic peptide of VP15 conferring antiviral activity against WSSV.

Highlights

  • White spot syndrome virus (WSSV) is one of the most devastating pathogens in penaeid shrimp and can cause massive damage in shrimp aquaculture industries

  • VP19, VP24, VP26, and VP28 were identified as envelope proteins, while VP15 and VP664 were identified as nucleocapsid-associated p­ roteins19,20

  • We attempted to determine the antigenic domain of the VP15 protein by in vivo animal experiments with M. japonicus using a purified truncated VP15 series and synthetic peptides derived from VP15 at the peptide level

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Summary

Introduction

White spot syndrome virus (WSSV) is one of the most devastating pathogens in penaeid shrimp and can cause massive damage in shrimp aquaculture industries. Among the four truncated constructs, ­VP15(26–57) provided a significant improvement in the shrimp survival rate after 20 days of viral infection. Most recombinant subunit vaccines investigated have been based on VP19, VP24, VP26, VP28, VP292, and VP466, and among them, VP28 has been widely ­studied33–37 These viral subunits were confirmed to improve the survival rate after challenging shrimp with WSSV. No protein crystal structure has been reported yet, and only one trial used VP15-based material (DNA vaccine encoding V­ P1542) for immunizing shrimp against WSSV. To further explore the mechanism of antigenicity involved, we investigated the possible interaction between VP15 and its host protein partner, a gC1qR homolog from M. japonicus (MjgC1qR), via GST pull-down assay

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