Development of a Recombinant Protein Vaccine Based on Cell-Free Protein Synthesis for Sevenband Grouper Epinephelus septemfasciatus Against Viral Nervous Necrosis
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초록
Sevenband grouper, Epinephelus septemfasciatus, is becoming an important aquaculture species in Korea. However, viral nervous necrosis disease is a large problem causing mass mortality in sevenband grouper aquaculture. Recombinant protein vaccines are one of the best methods to reduce these economic losses. However, the cell-based expression method mainly produces inclusion bodies and requires additional procedures. In this study, we expressed a recombinant viral coat protein of sevenband grouper nervous necrosis virus (NNV) using a cell-free protein synthesis system. The purified recombinant NNV coat protein (rNNV-CP) was injected into sevenband grouper at different doses followed by a NNV challenge. Nonimmunized fish in the first trial (20 μg/fish) began to die 5 days post-challenge and reached 70% cumulative mortality. In contrast, immunized fish also starting dying 5 days postchallenge but lower cumulative mortality (10%) was observed. Cumulative morality in the second trial with different doses (20, 4, and 0.8 μg/fish) was 10%, 40%, and 50%, respectively. These results suggest that rNNV-CP can effectively immunize sevenband grouper depending on the dose administered. This study provides a new approach to develop a recombinant vaccine against NNV infection for sevenband grouper.
Abstract
Sevenband grouper, Epinephelus septemfasciatus, is becoming an important aquaculture species in Korea. However, viral nervous necrosis disease is a large problem causing mass mortality in sevenband grouper aquaculture. Recombinant protein vaccines are one of the best methods to reduce these economic losses. However, the cell-based expression method mainly produces inclusion bodies and requires additional procedures. In this study, we expressed a recombinant viral coat protein of sevenband grouper nervous necrosis virus (NNV) using a cell-free protein synthesis system. The purified recombinant NNV coat protein (rNNV-CP) was injected into sevenband grouper at different doses followed by a NNV challenge. Nonimmunized fish in the first trial (20 μg/fish) began to die 5 days post-challenge and reached 70% cumulative mortality. In contrast, immunized fish also starting dying 5 days postchallenge but lower cumulative mortality (10%) was observed. Cumulative morality in the second trial with different doses (20, 4, and 0.8 μg/fish) was 10%, 40%, and 50%, respectively. These results suggest that rNNV-CP can effectively immunize sevenband grouper depending on the dose administered. This study provides a new approach to develop a recombinant vaccine against NNV infection for sevenband grouper.
참고문헌 (37)
- Brudeseth BE, "Status and future perspectives of vaccines for industrialised fin-fish farming", Fish Shellfish Immunol, 35, pp.1759~1768 (2013)
- Bundy BC, "Escherichia coli-based cell-free synthesis of virus-like particles", Biotechnol. Bioeng, 100, pp.28~37 (2008)
- Carlson ED, "Cellfree protein synthesis: applications come of age", Biotechnol. Adv, 30, pp.1185~1194 (2012)
- Cha SJ, "Phylogenetic analysis of betanodaviruses isolated from cultured fish in Korea", Dis. Aquat. Org, 77, pp.181~189 (2007)
- Choi YR, "Chromatographically-purified capsid proteins of red-spotted grouper nervous necrosis virus expressed in Saccharomyces cerevisiae form virus-like particles", Protein Expr. Purif, 89, pp.162~168 (2013)
- Clark TG, "Recombinant subunit vaccines: potential and constraints", Dev. Biol. (Basel), 121, pp.153~163 (2005)
- Goerke AR, "Development of cell-free protein synthesis platforms for disulfide bonded proteins", Biotechnol. Bioeng, 99, pp.351~367 (2008)
- Goshima N, "Human protein factory for converting the transcriptome into an in vitro-expressed proteome", Nat. Methods, 5, pp.1011~1017 (2008)
- Griffiths AD, "Directed evolution of an extremely fast phosphotriesterase by in vitro compartmentalization", EMBO J, 22, pp.24~35 (2003)
- Húsgard S, "Immune response to a recombinant capsid protein of striped jack nervous necrosis virus (SJNNV) in turbot Scophthalmus maximus and Atlantic halibut Hippoglossus hippoglossus, and evaluation of a vaccine against SJNNV", Dis. Aquat. Org, 45, pp.33~44 (2001)
- Kanter G, "Cell-free production of scFv fusion proteins: an efficient approach for personalized lymphoma vaccines", Blood, 109, pp.3393~3399 (2007)
- Katzen F, "The past, present and future of cell-free protein synthesis", Trends Biotechnol, 22, pp.150~156 (2005)
- 김춘섭, "능성어 양식장에서의 viral nervous necrosis (VNN) 발생양상", 한국어병학회지, 25(2), pp.111~116 (2012), 한국어병학회[DOI]
- Kim JH, "Infectious and Parasitic Diseases of Fish and Shellfish" (2006), Life Science Publishing Co
- Kim JO, "Complete genome sequence of nervous necrosis virus isolated from sevenband grouper (Epinephelus septemfasciatus) in South Korea", Genome Announc, 2, pp.e01264~e01214 (2014)
- Kim WS, "Change of viral hemorrhagic septicemia virus (VHSV) titer in olive flounder (Paralichthys olivaceus) following Poly(I:C) administration", Aquacul. Int, 22, pp.1175~1179 (2014)
- Lai YX, "Immune responses of orange-spotted grouper, Epinephelus coioides, against virus-like particles of betanodavirus produced in Escherichia coli", Vet. Immunol. Immunopathol, 157, pp.87~96 (2014)
- Liu W, "Immune response against grouper nervous necrosis virus by vaccination of virus-like particles", Vaccine, 24, pp.6282~6287 (2006)
- Lu MW, "The interferon response is involved in nervous necrosis virus acute and persistent infection in zebrafish infection model", Mol. Immunol, 45, pp.1146~1152 (2008)
- Lu MW, "Infection competition against grouper nervous necrosis virus by virus-like particles produced in Escherichia coli", J. Gen. Virol, 84, pp.1577~1582 (2003)
- Lorenzen N, "Molecular cloning and expression in Escherichia coli of the glycoprotein gene of VHS virus, and immunization of rainbow trout with the recombinant protein", J. Gen. Virol, 74, pp.623~630 (1993)
- Munday BL, "Betanodavirus infections of teleost fish: a review", J. Fish Dis, 25, pp.127~142 (2002)
- Munday BL, "Special topic review: nodaviruses as pathogens in larval and juvenile marine finfish", World J. Microbiol. Biotechnol, 13, pp.375~381 (1997)
- Nishizawa T, "Comparison of the coat protein genes of five fish nodaviruses, the causative agents of viral nervous necrosis in marine fish", J. Gen. Virol, 76, pp.1563~1569 (1995)
- Nishizawa T, "Polymerase chain reaction (PCR) amplification of RNA of striped jack nervous necrosis virus (SJNNV)", Dis. Aquat. Org, 18, pp.103~107 (1994)
- Nishizawa T, "Fish immunization using a synthetic double-stranded RNA Poly(I:C), an interferon inducer, offers protection against RGNNV, a fish nodavirus", Dis. Aquat. Org, 83, pp.115~122 (2009)
- Oh MJ, "Comparison of the coat protein gene of nervous necrosis virus (NNV) detected from marine fishes in Korea", J. World Aqua. Soc, 36, pp.223~227 (2005)
- Oh MJ, "Change in infectivity titre of nervous necrosis virus (NNV)in brain tissue of sevenband grouper, Epinephelus septemfasciatus, Thunberg, with poly(I:C) administration", J. Fish Dis, 36, pp.159~162 (2013)
- OIE, "Manual of Diagnostic Tests for Aquatic Animals. Viral Encephalopathy and Retinopathy", World Organisation for Animal Health, ~ (2009)
- Reed LJ, "A simple method of estimating fifty percent end points", Am. J. Hyg, 27, pp.493~497 (1938)
- Sohn SG, "Studies on the mass mortality of the cultured grouper, Epinephelus septemfasciatus", J. Fish Pathol, 4, pp.87~94 (1991)
- Sohn SG, "A fish nodavirus isolated from cultured sevenband grouper, Epinephelus septemfasciatus", J. Fish Pathol, 11, pp.97~104 (1998)
- Tanaka S, "Protective immunity of sevenband grouper, Epinephelus septemfasciatus Thunberg, against experimental viral nervous necrosis", J. Fish Dis, 24, pp.15~22 (2001)
- Thiery R, "Induction of a protective immune response against viral nervous necrosis in the European sea bass Dicentrarchus labrax by using betanodavirus virus-like particles", J. Virol, 80, pp.10201~10207 (2006)
- Wimmer, "The test-tube synthesis of a chemical called poliovirus", EMBO Rep, 7, pp.53~59 (2006)
- Yuasa K, "Immune response of humpback grouper, Cromileptes altivelis (Valenciennes), injected with the recombinant coat protein of betanodavirus", J. Fish Dis, 25, pp.53~56 (2002)
- Zawada J, "Maintaining rapid growth in moderate-density Escherichia coli fermentations", Biotechnol. Bioeng, 89, pp.407~415 (2005)