Generation of a novel live rabies vaccine strain with a high level of safety by introducing attenuating mutations in the nucleoprotein and glycoprotein

Keisuke Nakagawa, Kento Nakagawa, Tsutomu Omatsu, Yukie Katayama, Mami Oba, Hiromichi Mitake, Kazuma Okada, Satoko Yamaoka, Yasuhiro Takashima, Tatsunori Masatani, Kota Okadera, Naoto Ito, Tetsuya Mizutani, Makoto Sugiyama

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The current live rabies vaccine SAG2 is attenuated by only one mutation (Arg-to-Glu) at position 333 in the glycoprotein (G333). This fact generates a potential risk of the emergence of a pathogenic revertant by a back mutation at this position during viral propagation in the body. To circumvent this risk, it is desirable to generate a live vaccine strain highly and stably attenuated by multiple mutations. However, the information on attenuating mutations other than that at G333 is very limited. We previously reported that amino acids at positions 273 and 394 in the nucleoprotein (N273/394) (Leu and His, respectively) of fixed rabies virus Ni-CE are responsible for the attenuated phenotype by enhancing interferon (IFN)/chemokine gene expressions in infected neural cells. In this study, we found that amino acid substitutions at N273/394 (Phe-to-Leu and Tyr-to-His, respectively) attenuated the pathogenicity of the oral live vaccine ERA, which has a virulent-type Arg at G333. Then we generated ERA-N273/394-G333 attenuated by the combination of the above attenuating mutations at G333 and N273/394, and checked its safety. Similar to the ERA-G333, which is attenuated by only the mutation at G333, ERA-N273/394-G333 did not cause any symptoms in adult mice after intracerebral inoculation, indicating a low level of residual pathogenicity of ERA-N273/394-G333. Further examination revealed that infection with ERA-N273/394-G333 induces IFN-β and CXCL10 mRNA expressions more strongly than ERA-G333 infection in a neuroblastoma cell line. Importantly, we found that the ERA-N273/394-G333 stain has a lower risk for emergence of a pathogenic revertant than does the ERA-G333. These results indicate that ERA-N273/394-G333 has a potential to be a promising candidate for a live rabies vaccine strain with a high level of safety.

Original languageEnglish (US)
Pages (from-to)5622-5628
Number of pages7
JournalVaccine
Volume35
Issue number42
DOIs
StatePublished - Oct 9 2017

Fingerprint

Rabies Vaccines
nucleoproteins
Nucleoproteins
rabies
glycoproteins
Glycoproteins
vaccines
mutation
Safety
Mutation
live vaccines
interferons
Interferons
Virulence
pathogenicity
Vaccines
Rabies virus
amino acid substitution
chemokines
Amino Acid Substitution

Keywords

  • Attenuated live vaccine
  • Gene manipulation
  • Glycoprotein
  • Nucleoprotein
  • Rabies

ASJC Scopus subject areas

  • Molecular Medicine
  • Immunology and Microbiology(all)
  • veterinary(all)
  • Public Health, Environmental and Occupational Health
  • Infectious Diseases

Cite this

Generation of a novel live rabies vaccine strain with a high level of safety by introducing attenuating mutations in the nucleoprotein and glycoprotein. / Nakagawa, Keisuke; Nakagawa, Kento; Omatsu, Tsutomu; Katayama, Yukie; Oba, Mami; Mitake, Hiromichi; Okada, Kazuma; Yamaoka, Satoko; Takashima, Yasuhiro; Masatani, Tatsunori; Okadera, Kota; Ito, Naoto; Mizutani, Tetsuya; Sugiyama, Makoto.

In: Vaccine, Vol. 35, No. 42, 09.10.2017, p. 5622-5628.

Research output: Contribution to journalArticle

Nakagawa, K, Nakagawa, K, Omatsu, T, Katayama, Y, Oba, M, Mitake, H, Okada, K, Yamaoka, S, Takashima, Y, Masatani, T, Okadera, K, Ito, N, Mizutani, T & Sugiyama, M 2017, 'Generation of a novel live rabies vaccine strain with a high level of safety by introducing attenuating mutations in the nucleoprotein and glycoprotein', Vaccine, vol. 35, no. 42, pp. 5622-5628. https://doi.org/10.1016/j.vaccine.2017.08.050
Nakagawa, Keisuke ; Nakagawa, Kento ; Omatsu, Tsutomu ; Katayama, Yukie ; Oba, Mami ; Mitake, Hiromichi ; Okada, Kazuma ; Yamaoka, Satoko ; Takashima, Yasuhiro ; Masatani, Tatsunori ; Okadera, Kota ; Ito, Naoto ; Mizutani, Tetsuya ; Sugiyama, Makoto. / Generation of a novel live rabies vaccine strain with a high level of safety by introducing attenuating mutations in the nucleoprotein and glycoprotein. In: Vaccine. 2017 ; Vol. 35, No. 42. pp. 5622-5628.
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abstract = "The current live rabies vaccine SAG2 is attenuated by only one mutation (Arg-to-Glu) at position 333 in the glycoprotein (G333). This fact generates a potential risk of the emergence of a pathogenic revertant by a back mutation at this position during viral propagation in the body. To circumvent this risk, it is desirable to generate a live vaccine strain highly and stably attenuated by multiple mutations. However, the information on attenuating mutations other than that at G333 is very limited. We previously reported that amino acids at positions 273 and 394 in the nucleoprotein (N273/394) (Leu and His, respectively) of fixed rabies virus Ni-CE are responsible for the attenuated phenotype by enhancing interferon (IFN)/chemokine gene expressions in infected neural cells. In this study, we found that amino acid substitutions at N273/394 (Phe-to-Leu and Tyr-to-His, respectively) attenuated the pathogenicity of the oral live vaccine ERA, which has a virulent-type Arg at G333. Then we generated ERA-N273/394-G333 attenuated by the combination of the above attenuating mutations at G333 and N273/394, and checked its safety. Similar to the ERA-G333, which is attenuated by only the mutation at G333, ERA-N273/394-G333 did not cause any symptoms in adult mice after intracerebral inoculation, indicating a low level of residual pathogenicity of ERA-N273/394-G333. Further examination revealed that infection with ERA-N273/394-G333 induces IFN-β and CXCL10 mRNA expressions more strongly than ERA-G333 infection in a neuroblastoma cell line. Importantly, we found that the ERA-N273/394-G333 stain has a lower risk for emergence of a pathogenic revertant than does the ERA-G333. These results indicate that ERA-N273/394-G333 has a potential to be a promising candidate for a live rabies vaccine strain with a high level of safety.",
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AU - Katayama, Yukie

AU - Oba, Mami

AU - Mitake, Hiromichi

AU - Okada, Kazuma

AU - Yamaoka, Satoko

AU - Takashima, Yasuhiro

AU - Masatani, Tatsunori

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