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Dev Comp Immunol. 2017 Nov;76:56-64. doi: 10.1016/j.dci.2017.05.016. Epub 2017 May 22.

Vaccination with multimeric recombinant VP28 induces high protection against white spot syndrome virus in shrimp.

Author information

1
Shrimp-pathogen Interaction (SPI) laboratory, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Yothi Office, Rama VI Rd., Bangkok, 10400, Thailand.
2
Department of Environmental Science, Graduate School of Fisheries and Environmental Sciences, Nagasaki University, Nagasaki, Japan.
3
Centex Shrimp, Faculty of Science, Mahidol University, Rama VI Rd., Bangkok, 10400, Thailand; Department of Biotechnology, Faculty of Science, Mahidol University, Rama VI Rd., Bangkok, 10400, Thailand.
4
Shrimp-pathogen Interaction (SPI) laboratory, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Yothi Office, Rama VI Rd., Bangkok, 10400, Thailand; Centex Shrimp, Faculty of Science, Mahidol University, Rama VI Rd., Bangkok, 10400, Thailand; Departamento de Biotecnología y CienciasAlimentarias, InstitutoTecnológico de Sonora, Cd. Obregón, Sonora, Mexico.
5
Shrimp-pathogen Interaction (SPI) laboratory, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Yothi Office, Rama VI Rd., Bangkok, 10400, Thailand. Electronic address: Thailand.Kallaya@biotec.or.th.

Abstract

To improve the efficacy of WSSV protection, multimeric (tetrameric) recombinant VP28 (4XrVP28) was produced and tested in comparison with those of monomeric VP28 (1XrVP28). In vitro binding of either 1XrVP28 or 4XrVP28 to shrimp hemocyte surface was evident as early as 10 min after protein inoculation. Similar results were obtained in vivo when shrimp were injected with recombinant proteins that the proteins bound to the hemocyte surface could be detected since 5 min after injection. Comparison of the WSSV protection efficiencies of 1XrVP28 or 4XrVP28 were performed by injection the purified 1XrVP28 or 4XrVP28 (22.5 μg/shrimp) and WSSV inoculum (1000 copies/shrimp) into shrimp. At 10 dpi, while shrimp injected with WSSV inoculum reached 100% mortality, shrimp injected with 1XrVP28 + WSSV or 4XrVP28 + WSSV showed relative percent survival (RPS) of 67% and 81%, respectively. PCR quantification revealed high number of WSSV in the moribund shrimp of WSSV- and 1XrVP28+WSSV-injected group. In contrast, lower number of WSSV copies were found in the survivors both from 1XrVP28+WSSV- or 4XrVP28+WSSV- injected groups. Histopathological analysis demonstrated the WSSV infected lesions found in the moribund from WSSV-infected group and 1XrVP28+WSSV-injected group, but less or none in the survivors. ELISA demonstrated that 4XrVP28 exhibited higher affinity binding to rPmRab7, a WSSV binding protein essential for WSSV entry to the cell than 1XrVP28. Taken together, the protection against WSSV in shrimp could be improved by application of multimeric rVP28.

KEYWORDS:

Monomeric recombinant VP28 (1XrVP28); Penaeus monodon; RT; Tetrameric recombinant VP28 (4XrVP28); White spot syndrome virus (WSSV); day post injection; dpi; room temperature

PMID:
28545960
DOI:
10.1016/j.dci.2017.05.016
[Indexed for MEDLINE]

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