Clin Exp Vaccine Res.  2014 Jul;3(2):212-219. 10.7774/cevr.2014.3.2.212.

Inactivated genotype 1 Japanese encephalitis vaccine for swine

• Yang DK
• Nah JJ
• Kim HH
• Song JY

• KMID: 1730626
• DOI: http://doi.org/10.7774/cevr.2014.3.2.212

Abstract

PURPOSE
Japanese encephalitis is a reproductive disorder caused by Japanese encephalitis virus (JEV) in swine. Recent genotype (G) shift phenomenon (G3 to G1) in the Asia-wide has posed a challenge for proper prevention by the current vaccine strain. Thus, new kinds of JEV G1 vaccines with enhanced immunogenicity have been required for pigs.
MATERIALS AND METHODS
Recombinant porcine granulocyte monocyte-colony stimulating factor (reporGM-CSF) protein was expressed in Spodoptera frugiperda (Sf-9) cells using baculovirus expression system. Two kinds of trials with inactivated JEV vaccines containing IMS1313 adjuvant (Seppic, France) were prepared with or without reporGM-CSF protein. Safety and immunogenicity of the pigs inoculated with the JEV vaccines via intramuscular route was evaluated for 28 days after inoculation.
RESULTS
Mice, guinea pigs, and fattening pigs inoculated with the inactivated vaccine showed no signs for 14 and 21 days. Both hemagglutination inhibition and plaque reduction neutralizing antibody titers were significantly higher in pigs immunized with the vaccine containing reporGM-CSF protein after boosting. However, on the side of vaccine efficacy, most mice (87%) immunized with the inactivated JEV vaccine survived after virulent JEV challenge. Whereas the group with the vaccine containing reporGM-CSF protein showed lower protective effects than the vaccine alone for the biological activity of the GM-CSF depending on species specific.
CONCLUSION
Our data indicate that animals inoculated with the JEV vaccines was safe and pigs inoculated with inactivated JEV vaccine containing reporGM-CSF protein showed higher humoral immune responses than that of inactivated JEV vaccine without reporGM-CSF protein.

Keyword

Japanese encephalitis virus; Inactivated vaccine; GM-CSF; Swine

MeSH Terms

Animals
Antibodies, Neutralizing
Baculoviridae
Encephalitis Virus, Japanese
Encephalitis, Japanese*
Genotype*
Granulocyte-Macrophage Colony-Stimulating Factor
Granulocytes
Guinea Pigs
Hemagglutination
Immunity, Humoral
Mice
Spodoptera
Swine*
Vaccines
Antibodies, Neutralizing
Granulocyte-Macrophage Colony-Stimulating Factor
Vaccines

Figure

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  Fig. 1 Identification of Japanese encephalitis virus (JEV) strain (KV1899) for the inactivated JEV G1 vaccine by indirect fluorescent assay (×200). Specific cytoplasmic fluorescent was shown in the Vero cells infected with JEV.

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  Fig. 2 Cytopathic effect in Sf-9 cells infected with recombinant porcine granulocyte monocyte-colony stimulating factor baculovirus (A, ×200), and normal Sf-9 cells (B, ×200).

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  Fig. 3 Identification of porcine granulocyte monocyte-colony stimulating factor (GM-CSF) gene from cloned GM-CSF baculovirus. (A) M, 1 kb DNA ladder; lane 1, porcine GM-CSF gene amplified with specific primers; lane 2, normal Sf-9 cell; lane 3, Sf-9 cells inoculated with recombinant porcine GM-CSF (reporGM-CSF) baculovirus. The expression of the porcine GM-CSF protein using baculovirus vector system. (B) M, protein marker; lane 1, normal Sf-9 cell; lane 2, supernatant of Sf-9 cells inoculated with reporGM-CSF baculovirus; lane 3, lysate of Sf-9 cell inoculated with reporGM-CSF baculovirus.

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  Fig. 4 Antibody responses following vaccination with inactivated Japanese encephalitis virus (JEV) G1 vaccine containing recombinant porcine granulocyte monocyte-colony stimulating factor (reporGM-CSF) protein or not. The pigs were bled at the day of boosting (pre-boost), and 14 days post-boosting for hemagglutination inhibition (HI) titers (A) and plaque reduction neutralization test (PRNT) titers (B). The HI and PRNT titers were log2 converted and reported as the geometric mean. Data were shown as expressed mean±standard deviation of 8 pigs. NS, not significant. *p < 0.05.

• 

  Fig. 5 Survival rate of the immunized mice after challenge. After immunization with the inactivated Japanese encephalitis virus (JEV) vaccine with or without recombinant porcine granulocyte monocyte-colony stimulating factor (reporGM-CSF) protein, mice were challenged with 100 LD50 of virulent JEV.

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