Oral Vaccination of Baculovirus-Expressed VP28 Displays Enhanced Protection against White Spot Syndrome Virus in Penaeus monodon

White Spot Syndrome Virus (WSSV) is an infectious pathogen of shrimp and other crustaceans, and neither effective vaccines nor adequate treatments are currently available. WSSV is an enveloped dsDNA virus, and one of its major envelope proteins, VP28, plays a pivotal role in WSSV infection. In an attempt to develop a vaccine against WSSV, we inserted the VP28 gene into a baculovirus vector tailored to express VP28 on the baculovirus surface under the WSSV ie1 promoter (Bac-VP28). The Bac-VP28 incorporated abundant quantity (65.3 µg/ml) of VP28. Shrimp were treated by oral and immersion vaccination with either Bac-VP28 or wild-type baculovirus (Bac-wt). The treatment was followed by challenge with WSSV after 3 and 15 days. Bac-VP28 vaccinated shrimp showed significantly higher survival rates (oral: 81.7% and 76.7%; immersion: 75% and 68.4%) than Bac-wt or non-treated shrimp (100% mortality). To verify the protective effects of Bac-VP28, we examined in vivo expression of VP28 by immunohistochemistry and quantified the WSSV copy number by qPCR. In addition to that, we quantified the expression levels shrimp genes LGBP and STAT by real-time RT-PCR from the samples obtained from Bac-VP28 vaccinated shrimp at different duration of vaccine regime. Our findings indicate that oral vaccination of shrimp with Bac-VP28 is an attractive preventative measure against WSSV infection that can be used in the field.

[1]  G. Vasta,et al.  New Directions in Invertebrate Immunology , 1996 .

[2]  J. Hoffmann,et al.  Innate immunity in higher insects. , 1996, Current opinion in immunology.

[3]  C. Lo,et al.  Identification of the Nucleocapsid, Tegument, and Envelope Proteins of the Shrimp White Spot Syndrome Virus Virion , 2006, Journal of Virology.

[4]  Zi-rong Xu,et al.  [Effect of white spot syndrome virus envelope protein Vp28 expressed in silkworm (Bombyx mori) pupae on disease resistence in Procambarus clarkii]. , 2005, Shi yan sheng wu xue bao.

[5]  D. Amend Potency testing of fish vaccines , 1980 .

[6]  H. Qiu,et al.  Efficient gene delivery into mammalian cells mediated by a recombinant baculovirus containing a whispovirus ie1 promoter, a novel shuttle promoter between insect cells and mammalian cells. , 2007, Journal of biotechnology.

[7]  T. Flegel Update on viral accommodation, a model for host-viral interaction in shrimp and other arthropods. , 2007, Developmental and comparative immunology.

[8]  S. K. Kim,et al.  Protection of shrimp (Penaeus chinensis) against white spot syndrome virus (WSSV) challenge by double-stranded RNA. , 2007, Fish & shellfish immunology.

[9]  R. Huang,et al.  A novel envelope protein involved in White spot syndrome virus infection. , 2005, The Journal of general virology.

[10]  A. S. S. Hameed,et al.  Oral Administration of Bacterially Expressed VP28dsRNA to Protect Penaeus monodon from White Spot Syndrome Virus , 2008, Marine Biotechnology.

[11]  A. S. S. Hameed,et al.  Experimental infection of twenty species of Indian marine crabs with white spot syndrome virus (WSSV). , 2003, Diseases of aquatic organisms.

[12]  T. Nishizawa,et al.  Vaccination trials with Penaeus japonicus to induce resistance to white spot syndrome virus , 2004 .

[13]  Jianfang Ning,et al.  Oral delivery of DNA vaccine encoding VP28 against white spot syndrome virus in crayfish by attenuated Salmonella typhimurium. , 2009, Vaccine.

[14]  Norbert Perrimon,et al.  The roles of JAK/STAT signaling in Drosophila immune responses , 2004, Immunological reviews.

[15]  S. R. Kumar,et al.  Neutralization of white spot syndrome virus of shrimp by antiserum raised against recombinant VP28 , 2006 .

[16]  C. Lo,et al.  PmRab7 Is a VP28-Binding Protein Involved in White Spot Syndrome Virus Infection inShrimp , 2006, Journal of Virology.

[17]  D. K. Kim,et al.  Shrimp protected from WSSV disease by treatment with egg yolk antibodies (IgY) against a truncated fusion protein derived from WSSV , 2004 .

[18]  A. Rocha,et al.  DNA vaccination by immersion and ultrasound to trout viral haemorrhagic septicaemia virus. , 2001, Vaccine.

[19]  G. Kurath,et al.  Fish DNA vaccine against infectious hematopoietic necrosis virus: efficacy of various routes of immunisation. , 2000, Fish & shellfish immunology.

[20]  A. S. S. Hameed,et al.  An investigation of Artemia as a possible vector for white spot syndrome virus (WSSV) transmission to Penaeus indicus , 2002 .

[21]  Sudhir Kumar,et al.  DNA vaccines encoding viral envelope proteins confer protective immunity against WSSV in black tiger shrimp. , 2007, Vaccine.

[22]  C. Lo,et al.  White Spot Syndrome Virus Annexes a Shrimp STAT To Enhance Expression of the Immediate-Early Gene ie1 , 2006, Journal of Virology.

[23]  J. Kwang,et al.  Localization of VP28 on the baculovirus envelope and its immunogenicity against white spot syndrome virus in Penaeus monodon. , 2009, Virology.

[24]  A. S. S. Hameed,et al.  White spot syndrome virus (WSSV) in two species of freshwater crabs (Paratelphusa hydrodomous and P. pulvinata) , 2001 .

[25]  J. Vlak,et al.  Protection of Penaeus monodon against White Spot Syndrome Virus by Oral Vaccination , 2004, Journal of Virology.

[26]  J. Hoffmann,et al.  The Jak-STAT signaling pathway is required but not sufficient for the antiviral response of drosophila , 2005, Nature Immunology.

[27]  I. Hirono,et al.  Polymerase chain reaction (PCR) amplification of bacilliform virus (RV‐PJ) DNA in Penaeus japonicus Bate and systemic ectodermal and mesodermal baculovirus (SEMBV) DNA in Penaeus monodon Fabricius , 1996 .

[28]  Y. Nishida,et al.  Transcriptional Regulation of the Drosophila rafProto-oncogene by Drosophila STAT during Development and in Immune Response* , 2000, The Journal of Biological Chemistry.

[29]  Jyh-Ming Tsai,et al.  Genomic and Proteomic Analysis of Thirty-Nine Structural Proteins of Shrimp White Spot Syndrome Virus , 2004, Journal of Virology.

[30]  Wei Yu Chen,et al.  Wssv Infection Activates Stat in Shrimp , 2008 .

[31]  Y. Lai,et al.  Avian influenza virus hemagglutinin display on baculovirus envelope: cytoplasmic domain affects virus properties and vaccine potential. , 2007, Molecular therapy : the journal of the American Society of Gene Therapy.

[32]  R. Knobler,et al.  Western and dot immunoblotting analysis of viral antigens and antibodies: Application to murine hepatitis virus , 1984, Journal of Immunological Methods.

[33]  T. Nishizawa,et al.  A time-course study on the resistance of Penaeus japonicus induced by artificial infection with white spot syndrome virus. , 2002, Fish & shellfish immunology.

[34]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[35]  R. Sudhakaran,et al.  Experimental transmission and tissue tropism of white spot syndrome virus (WSSV) in two species of lobsters, Panulirus homarus and Panulirus ornatus. , 2006, Journal of invertebrate pathology.

[36]  Xin Ma,et al.  The interaction of white spot syndrome virus envelope protein VP28 with shrimp Hsc70 is specific and ATP-dependent. , 2009, Fish & shellfish immunology.

[37]  J. Vlak,et al.  Protection of Penaeus monodon against white spot syndrome virus using a WSSV subunit vaccine. , 2004, Fish & shellfish immunology.

[38]  F. Kafatos,et al.  Anopheles gambiae Ag‐STAT, a new insect member of the STAT family, is activated in response to bacterial infection , 1999, The EMBO journal.

[39]  A. Pain,et al.  The Lipopolysaccharide and β-1,3-Glucan Binding Protein Gene Is Upregulated in White Spot Virus-Infected Shrimp (Penaeus stylirostris) , 2002, Journal of Virology.

[40]  J. Vlak,et al.  White spot syndrome virus envelope protein VP28 is involved in the systemic infection of shrimp. , 2001, Virology.

[41]  Zi-rong Xu,et al.  The efficacy of recombinant vaccines against white spot syndrome virus in Procambarus clarkii. , 2006, Immunology letters.

[42]  T. Nishizawa,et al.  Quasi-immune response of Penaeus japonicus to penaeid rod-shaped DNA virus (PRDV). , 2000, Diseases of aquatic organisms.

[43]  D. Lightner,et al.  Quantitative real time PCR for the measurement of white spot syndrome virus in shrimp , 2002 .