Yang Dong-Kun, Nakagawa Keisuke, Ito Naoto, Kim Ha-Hyun, Hyun Bang-Hun, Nah Jin-Ju, Sugiyama Makoto, Song Jae-Young
Viral Disease Division, Animal and Plant Quarantine Agency, MAFRA, Anyang, Korea.
The United Graduated School of Veterinary Science, Gifu University, Gifu, Japan.
Clin Exp Vaccine Res. 2014 Jul;3(2):176-84. doi: 10.7774/cevr.2014.3.2.176. Epub 2014 Jun 20.
New alternative bait rabies vaccines applicable to pet dogs and wild animals are needed to eradicate rabies in Korea. In this study, recombinant rabies virus, ERAG3G strain was constructed using reverse genetic system and the safety, efficacy and immunogenicity of the ERAG3G strain was evaluated in mice and dogs.
Using the full-length genome mutated amino acid at position 333 of glycoprotein of rabies virus (RABV) and helper plasmids, the ERAG3G strain was rescued in BHK/T7-9 cells successfully. Mice were inoculated with the ERAG3G strain for safety and efficacy. Safety and immunogenicity of the dog inoculated with the ERAG3G strain (1 mL, 10(8.0) FAID50/mL) via intramuscular route was evaluated for 28 days after inoculation.
The ERAG3G strain rescued by reverse genetic system was propagated well in the mouse neuroblastoma cells revealing titer of 10(8.5) FAID50/mL and was not pathogenic to 4- or 6-week-old mice that received by intramuscular or intracranical route. Immunization with the ERAG3G strain conferred complete protection from lethal RABV in mice. Dogs inoculated with the vaccine candidate via intramuscular route showed high neutralizing antibody titer ranging from 2.62 to 23.9 IU/mL at 28 days postinoculation.
Our findings suggest that the ERAG3G strain plays an important role in inducing protective efficacy in mice and causes to arise anti-rabies neutralizing antibody in dogs.
为在韩国根除狂犬病，需要适用于宠物狗和野生动物的新型替代诱饵狂犬病疫苗。在本研究中，使用反向遗传系统构建了重组狂犬病病毒ERAG3G株，并在小鼠和犬中评估了ERAG3G株的安全性、有效性和免疫原性。
利用狂犬病病毒（RABV）糖蛋白第333位氨基酸突变的全长基因组和辅助质粒，在BHK/T7-9细胞中成功拯救出ERAG3G株。给小鼠接种ERAG3G株以评估安全性和有效性。通过肌肉途径给犬接种ERAG3G株（1 mL，10(8.0) FAID50/mL），接种后28天评估其安全性和免疫原性。
通过反向遗传系统拯救的ERAG3G株在小鼠神经母细胞瘤细胞中生长良好，滴度为10(8.5) FAID50/mL，对通过肌肉或颅内途径接种的4周龄或6周龄小鼠无致病性。用ERAG3G株免疫可使小鼠完全免受致死性RABV感染。通过肌肉途径接种候选疫苗的犬在接种后28天显示出高中和抗体滴度，范围为2.62至23.9 IU/mL。
我们的研究结果表明ERAG3G株在诱导小鼠的保护效力方面发挥重要作用，并能使犬产生抗狂犬病中和抗体。
