Clin Exp Vaccine Res.  2015 Jul;4(2):189-194. 10.7774/cevr.2015.4.2.189.

Application of recombinant adenoviruses expressing glycoprotein or nucleoprotein of rabies virus to Korean raccoon dogs

• Choi J
• Yang DK
• Kim HH
• Jo HY
• Choi SS
• Kim JT
• Cho IS
• Kim HW

• KMID: 1965404
• DOI: http://doi.org/10.7774/cevr.2015.4.2.189

Abstract

PURPOSE
A new rabies vaccine for animals, including raccoon dogs, in Korea is needed to eradicate rabies infection. In this study, we constructed two recombinant adenoviruses expressing the glycoprotein or nucleoprotein of the rabies virus (RABV). We then investigated the safety and immunogenicity of these strains in raccoon dogs, depending on inoculation route.
MATERIALS AND METHODS
Recombinant adenoviruses expressing the glycoprotein (Ad-0910G) or nucleoprotein (Ad-0910N) of rabies were constructed in 293A cells using an adenoviral system. One-year-old raccoon dogs underwent intramuscular (IM) inoculation or oral administration of the recombinant Ad-0910G and Ad-0910N. Clinical symptoms were observed and virus-neutralizing antibodies (VNA) against RABV were measured at 0, 2, 4, and 6 weeks after the immunization. Raccoons were considered positive if VNA titers were > or = 0.1 IU/mL.
RESULTS
Raccoon dogs inoculated with the combined Ad-0910G and Ad-0910N virus via the IM route did not exhibit any clinical sign of rabies during the observation period. All raccoon dogs (n = 7) immunized IM had high VNA titers, ranging from 0.17 to 41.6 IU/mL at 2 weeks after inoculation, but 70% (7/10) of raccoon dogs administered viruses via the oral route responded by 6 weeks after administration against RABV.
CONCLUSION
Raccoon dogs inoculated with Ad-0910G and Ad-0910N viruses showed no adverse effects. Immunization with the combined Ad-0910G and Ad-0910N strains may play an important role in inducing VNA against RABV in raccoon dogs.

Keyword

Adenovirus type 5; Rabies virus; Raccoon dogs

MeSH Terms

Adenoviridae*
Administration, Oral
Animals
Antibodies
Glycoproteins*
Immunization
Korea
Nucleoproteins*
Rabies Vaccines
Rabies virus*
Rabies*
Raccoon Dogs*
Raccoons*
Antibodies
Glycoproteins
Nucleoproteins
Rabies Vaccines

Figure

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  Fig. 1 The recombinant adenoviruses expressing glycoprotein or nucleoprotein of rabies virus were constructed by the ViraPower Adenoviral System. Each adenoviral expression clone was generated by performing an attL×attR (LR) recombination reaction between the entry plasmid and replication-deficient, E1-deleted, human adenovirus (Ad) type 5, pAd-DEST vector. Two generated-adenoviral expression clones (pAd-G and pAd-N) were digested with PacI, and then transfected into 293A cells. The viral titer of Ad-0910G and Ad-0910N strain propagated in 293A cells reached 108.0 TCID50/mL. After fixing the Ad-0910G or Ad-0910N infected cells with cold acetone, fluorescent assay (FA) was conducted with monoclonal antibodies against rabies virus.

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