A Novel System for Constructing a Recombinant Highly-Attenuated Vaccinia Virus Strain (LC16m8) Expressing Foreign Genes and Its Application for the Generation of LC16m8-Based Vaccines against Herpes Simplex Virus 2

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  • Omura Natsumi
    Department of Virology 1, National Institute of Infectious Diseases Department of Life Science and Medical Bioscience, Waseda University
  • Yoshikawa Tomoki
    Department of Virology 1, National Institute of Infectious Diseases
  • Fujii Hikaru
    Department of Virology 1, National Institute of Infectious Diseases
  • Shibamura Miho
    Department of Virology 1, National Institute of Infectious Diseases
  • Inagaki Takuya
    Department of Virology 1, National Institute of Infectious Diseases Department of Life Science and Medical Bioscience, Waseda University
  • Kato Hirofumi
    Department of Virology 1, National Institute of Infectious Diseases
  • Egawa Kazutaka
    Department of Virology 1, National Institute of Infectious Diseases
  • Harada Shizuko
    Department of Virology 1, National Institute of Infectious Diseases
  • Yamada Souichi
    Department of Virology 1, National Institute of Infectious Diseases
  • Takeyama Haruko
    Department of Life Science and Medical Bioscience, Waseda University
  • Saijo Masayuki
    Department of Virology 1, National Institute of Infectious Diseases

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<p>A novel system was developed for generating highly attenuated vaccinia virus LC16m8 (m8, third-generation smallpox vaccine) that expresses foreign genes. The innovations in this system are its excisable selection marker, specificity of the integration site of a gene of interest, and easy identification of clones with a fluorescent signal. Using this system, recombinant m8s, which expressed herpes simplex virus 2 (HSV-2) glycoprotein B (gB)-, gD-, or both gB and gD (gB + gD), were generated, and their efficacy was evaluated. First, the induction of a specific IgG against these HSV-2 glycoproteins in mice infected with one of these recombinant m8s was confirmed by an immunofluorescent assay. Next, mice preinfected with one of the recombinant m8s were infected with HSV-2 at a lethal dose to examine the vaccine efficacy. The fatality rate among the mice preinfected with either the recombinant gB + gD- or gD-expressing m8 significantly decreased in comparison with the control. The survival rate in male and female mice preinfected with either the recombinant gB + gD- or gD-expressing m8 increased to 100% and 60%, respectively, while most of the control mice died. In summary, this new system may be applicable to creation of a novel m8-based vaccine.</p>

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