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前園　佳祐 (マエゾノ　ケイスケ)

獣医学研究院　獣医学部門　衛生学分野助教
人獣共通感染症国際共同研究所助教

研究者基本情報

■ 学位
  • 学士（獣医学）, 2021年03月
■ URL
researchmap URLホームページURL■ ID 各種
研究者番号
  • 61008497
ORCID IDJ-Global ID■ 担当教育組織

経歴

■ 経歴
経歴
  • 2024年06月 - 現在
    北海道大学 大学院獣医学研究院 獣医学部門 衛生学分野 公衆衛生学教室, 助教
  • 2024年04月 - 2026年03月
    独立行政法人日本学術振興会, 特別研究員DC2
  • 2021年04月 - 2022年03月
    森永乳業（株）, 研究本部
学歴
  • 2022年04月 - 2024年05月, 北海道大学, 大学院獣医学研究院
  • 2015年04月 - 2021年03月, 北海道大学, 獣医学部

研究活動情報

■ 論文
  • Molecular evolution of Hokkaido virus, a genotype of Orthohantavirus puumalaense, among Myodes rodents.
    Duong Thi Ngoc Thuy; Michihito Sasaki; Yasuko Orba; Passawat Thammahakin; Keisuke Maezono; Shintaro Kobayashi; Hiroaki Kariwa
    Virology, 597, 110168, 110168, 2024年09月, ［国際誌］
    英語, 研究論文（学術雑誌）, Viruses in the genus Orthohantavirus within the family Hantaviridae cause human hantavirus infections and represent a threat to public health. Hokkaido virus (HOKV), a genotype of Orthohantavirus puumalaense (Puumala virus; PUUV), was first identified in Tobetsu, Hokkaido, Japan. Although it is genetically related to the prototype of PUUV, the evolutionary pathway of HOKV is unclear. We conducted a field survey in a forest in Tobetsu in 2022 and captured 44 rodents. Complete coding genome sequences of HOKVs were obtained from five viral-RNA-positive rodents (four Myodes rufocanus bedfordiae and one Apodemus speciosus). Phylogenetic analysis revealed a close relationship between the phylogenies and geographical origins of M. rufocanus-related orthohantaviruses. Comparison of the phylogenetic trees of the S segments of orthohantaviruses and the cytochrome b genes of Myodes species suggested that Myodes-related orthohantaviruses evolved in Myodes rodent species as a result of genetic isolation and host switching.
  • Rab27a promotes degradation of West Nile virus E protein in the lysosome.
    Shintaro Kobayashi; Seira Kawai; Yukine Fukuda; Haruto Eguchi; Keisuke Maezono; Passawat Thammahakin; Hirofumi Sawa; Hiroaki Kariwa
    iScience, 27, 4, 109539, 109539, 2024年04月19日, ［国際誌］
    英語, 研究論文（学術雑誌）, Rab27a, a Rab family small GTPases, plays an important role in the trafficking and secretion of the intracellular proteins and has been reported to promote various viral multiplication. However, whether Rab27a is involved in West Nile virus (WNV) multiplication is unknown. This study examined the ability of Rab27a to suppress WNV multiplication. The inhibition of Rab27a expression increased viral multiplication and the intracellular levels of WNV structural proteins, E and prM proteins. Rab27a partially colocalized with E protein, mainly in the perinuclear region, while inhibition of Rab27a expression resulted in diffuse subcellular localization of E protein. In addition, some of the perinuclear E protein colocalized with the lysosomal marker LAMP1, and inhibition of lysosomal acidification increased intracellular levels of Rab27a and E proteins. These observations suggested that Rab27a inhibits WNV multiplication by inducing the degradation of viral protein in lysosomes.
  • Ubiquitin accumulation induced by the finger and palm sub-domains of NS5 modulates the replication of West Nile virus.
    Shintaro Kobayashi; Ryoko Kawakami; Chisaki Takeda; Keisuke Maezono; Passawat Thammahakin; Haruto Eguchi; Bernard M Hang'ombe; Yasuko Orba; Hirofumi Sawa; Kentaro Yoshii; Hiroaki Kariwa
    Virology, 588, 109902, 109902, 2023年11月, ［国際誌］
    英語, 研究論文（学術雑誌）, West Nile virus (WNV) causes encephalitis in human and animals. WNV is phylogenetically classified into at least five distinct genetic lineages with different pathogenicity. The pathogenesis of West Nile encephalitis is affected by ubiquitin accumulation in infected cells, but the mechanism is unknown. In this study, the association between ubiquitin accumulation and WNV pathogenicity was investigated. Ubiquitin accumulation was detected in cells infected with NY99 strain belonging to lineage-1, but not FCG and Zmq16 strains belonging to lineage-2. Substitution of the Finger and Palm sub-domains of NS5 from lineage-1 to -2 decreased ubiquitin accumulation and viral replication. Furthermore, the survival rate was increased, and viral replication and ubiquitin accumulation in the brain were attenuated, in mice inoculated with the substituted WNV compared with lineage-1 WNV. Therefore, the intracellular ubiquitin accumulation induced by the Finger and Palm sub-domains of NS5 is linked to the differences in pathogenicity among WNV lineages.
  • Detection of disease-associated microglia among various microglia phenotypes induced by West Nile virus infection in mice.
    Passawat Thammahakin; Keisuke Maezono; Naoya Maekawa; Hiroaki Kariwa; Shintaro Kobayashi
    Journal of neurovirology, 29, 4, 367, 375, 2023年08月, ［国際誌］
    英語, 研究論文（学術雑誌）, West Nile virus (WNV) has emerged as a significant cause of viral encephalitis in humans and horses. However, the pathogenesis of the West Nile encephalitis remains unclear. Microglia are activated by WNV infection, and the pathogenic involvement of their phenotypes is controversial. In this study, we examined the diversity of microglia phenotypes caused by WNV infection by assessing various microglia markers and identified disease-associated microglia in WNV-infected mouse brain tissue. Cells positive for general microglia markers such as Iba1, P2RY12, or TMEM119 were detected in the control and WNV-infected brain tissue. The morphology of the positive cells in brain tissue infected by WNV was different from that of control brain tissue, indicating that WNV infection induced activation of microglia. The activated microglia were classified into various phenotypes by investigation of specific marker expression. Among the activated microglia, disease-associated microglia that were positive for CD11c and weakly positive for TMEM119 were detected close to the WNV-infected cells. These results indicate that WNV infection induces activation of diverse microglia phenotypes and that disease-associated microglia may be associated with the pathogenicity of WNV infection in the mouse brain.
  • Development of recombinant West Nile virus expressing mCherry reporter protein.
    Shintaro Kobayashi; Yukine Fukuda; Kentaro Yoshii; Passawat Thammahakin; Keisuke Maezono; Luděk Eyer; Daniel Růžek; Hiroaki Kariwa
    Journal of virological methods, 317, 114744, 114744, 2023年07月, ［国際誌］
    英語, 研究論文（学術雑誌）, West Nile virus (WNV) is transmitted to humans and animals by a mosquito and enters the central nervous system, leading to lethal encephalitis. Reporter viruses expressing fluorescent proteins enable detection of infected cells in vitro and in vivo, facilitating evaluation of the dynamics of viral infection, and the development of diagnostic or therapeutic methods. In this study, we developed a method for production of a recombinant replication-competent WNV expressing mCherry fluorescent protein. The expression of mCherry was observed in viral antigen-positive cells in vitro and in vivo, but the growth of the reporter WNV was reduced as compared to the parental WNV. The expression of mCherry was stable during 5 passages in reporter WNV-infected culture cells. Neurological symptoms were observed in mice inoculated intracranially with the reporter WNV. The reporter WNV expressing mCherry will facilitate research into WNV replication in mouse brains.
  • Development of a highly specific serodiagnostic ELISA for West Nile virus infection using subviral particles.
    Keisuke Maezono; Shintaro Kobayashi; Koshiro Tabata; Kentaro Yoshii; Hiroaki Kariwa
    Scientific reports, 11, 1, 9213, 9213, 2021年04月28日, ［国際誌］
    英語, 研究論文（学術雑誌）, West Nile virus (WNV), a member of the Japanese encephalitis virus (JEV) serocomplex group, causes lethal encephalitis in humans and horses. Because serodiagnosis of WNV and JEV is hampered by cross-reactivity, the development of a simple, secure, and WNV-specific serodiagnostic system is required. The coexpression of prM protein and E protein leads to the secretion of subviral particles (SPs). Deletion of the C-terminal region of E protein is reported to affect the production of SPs by some flaviviruses. However, the influence of such a deletion on the properties and antigenicity of WNV E protein is unclear. We analyzed the properties of full-length E protein and E proteins lacking the C-terminal region as novel serodiagnostics for WNV infection. Deletion of the C-terminal region of E protein suppressed the formation of SPs but did not affect the production of E protein. The sensitivity of an enzyme-linked immunosorbent assay (ELISA) using the full-length E protein was higher than that using the truncated E proteins. Furthermore, in the ELISA using full-length E protein, there was little cross-reactivity with anti-JEV antibodies, and the sensitivity was similar to that of the neutralization test.
■ その他活動・業績
■ 共同研究・競争的資金等の研究課題