Researcher basic information

• 博士 (獣医学), 北海道大学, Dec. 2015

• https://researchmap.jp/hiono?lang=en

• https://jglobal.jst.go.jp/detail?JGLOBAL_ID=201601000703654758

• 00775819

• 201601000703654758

• lectin
• glycan
• influenza

• Life Science, Veterinary medical science
• Life Science, Functional biochemistry, 糖鎖生物学
• Life Science, Virology

• Bachelor's degree program, School of Veterinary Medicine
• Doctoral (PhD) degree program, Graduate School of Infectious Diseases

Research activity information

• Nov. 2024, 日本農学会, 日本農学進歩賞
  鳥インフルエンザウイルスの生態と宿主域に関する研究
• Oct. 2024, Asian Association of Veterinary Schools, Kei-ichiro Maeda Memorial T&C Award
  Glycoscientific approach for the understanding of life-cycle and host range of influenza virus and SARS-CoV-2
• Sep. 2019, 日本獣医学会, 獣医学奨励賞
  鳥インフルエンザウイルスの異種宿主間伝播機構解明に向けたヘマグルチニン分子の糖質科学的解析
• Apr. 2015, 9th International Symposium on Avian Influenza, Student Poster Award
  Takahiro Hiono
• Mar. 2013, 日本獣医学会微生物分科会, 第4回若手奨励賞
  日尾野隆大

• Length and density of α2-3 sialyllactose-containing chains on glycopolymers determine receptor binding of avian influenza viruses
  Daiki Kobayashi; Takahiro Hiono; Ryota Adachi; Manabu Igarashi; Takahiko Matsushita; Norikazu Isoda; Yoshihiro Sakoda; Koji Matsuoka
  Archives of Virology, 171, 3, Springer Science and Business Media LLC, 26 Feb. 2026
  Scientific journal
• Neuraminidase of influenza A viruses induces global desialylation of host cells via its intracellular function.
  Daiki Kobayashi; Takahiro Hiono; Norikazu Isoda; Yoshihiro Sakoda
  Microbiology spectrum, e0332825, 18 Feb. 2026, [International Magazine]
  English, Scientific journal, The neuraminidase protein (NA) of influenza A viruses (IAVs) plays a role in the release of viruses from infected cells. NA on viral particles hydrolyzes sialylated glycans on the cell surface for viral budding. However, the maturation and intracellular functions of NA are poorly understood. To investigate how NA functions intracellularly, glycans displayed on IAV-infected cells were profiled by lectins, and global glycome alterations, accompanied by exposed terminal galactose and glycan recapping with α1-2 fucose, were found in the virus-infected cells. Since α1-2 fucosyltransferases are localized in the Golgi, these unique structures suggest a potential NA function in the IAV-infected cells. Functional analyses using antivirals and NA-expressing cells indicate that intracellular NA function is necessary for the glycome alterations. Time-course analyses in IAV-infected cells revealed that global desialylation and α1-2 fucosylation could be observed 5 h post-inoculation, corresponding to the timeframe of viral protein expression. These observations provide a novel theory of NA functions that NA obtains its enzymatic activity intracellularly before virus assembly and serves desialylated glycans for competitive glycosyltransferases, including α1-2 fucosyltransferases, as their acceptors, resulting in glycan recapping with α1-2 fucose. Hence, intracellular NA blocks the re-sialylation of glycans, promoting efficient virus release from infected cells by inhibiting the interaction between progeny virions and sialosides. This study further demonstrated the potential NA functions of limiting secondary IAV infection. These findings provide insights into the evolutionary strategies of IAVs for shaping the strict window of superinfection by NA functions under a balance between successful replication and reassortment.IMPORTANCEInfluenza A viruses (IAVs) exploit glycans for their replication cycle. The hemagglutinin protein uses sialic acid for viral attachment, and the neuraminidase protein (NA) hydrolyzes sialosides for virus release. However, the intracellular functions of NA are not well understood. This study demonstrated that intracellular NA induces global desialylation and glycan recapping with unique structures in IAV-infected cells. This suggests a novel mode of NA function during the IAV lifecycle, where virus particles are ready to be released at the assembly, and NAs no longer need to hydrolyze the sialic acids upon egress from the cells. Therefore, the present study provides novel and significant insights into the fundamental understanding of the lifecycle of IAV. Furthermore, as NA is a primary target for anti-influenza drugs, understanding the mechanism of intracellular NA function may also support the development of antivirals.
• Comparative evaluation of commercial enzyme-linked immunosorbent assay kits for antibody monitoring of classical swine fever virus in Japanese pig herds: performance assessment of domestic and foreign kits.
  Miki Koyasu; Keisuke Kuwata; Shuko Inoha; Yoko Kimura; Kaoru Hatate; Daiki Kobayashi; Takahiro Hiono; Norikazu Isoda; Yoshihiro Sakoda
  The Journal of veterinary medical science, 88, 2, 347, 354, 01 Feb. 2026, [Domestic magazines]
  English, Scientific journal, Classical swine fever (CSF) is a highly contagious disease in pigs, and vaccination with antibody monitoring is critical for its prevention. In this study, the effectiveness of two enzyme-linked immunosorbent assay (ELISA) kits for antibody detection against CSF virus (CSFV)-an indirect ELISA kit authorized in Japan in 2001 (N-kit) and a competitive ELISA kit additionally authorized in 2024 (I-kit)-was compared. For each ELISA kit, detection accuracy in terms of sensitivity, specificity and agreement rate, and quantitative accuracy were evaluated based on neutralizing antibody titers determined by serum neutralization tests. In addition, the impact of serum heat-inactivation at 56°C for 30 min on ELISA results was assessed. The results indicated that the I-kit showed the highest sensitivity and agreement rate in detection accuracy, whereas the N-kit showed the highest quantitative accuracy. Although blocking rates of the I-kit increased after heat-inactivation, high correlation rates between treated and untreated samples were confirmed for both kits, suggesting that heat-inactivation does not affect the final interpretation of the test results. These findings demonstrated that the I-kit is suitable for initial antibody screening in pigs due to its higher sensitivity, whereas the N-kit provides better quantitative accuracy, making it preferable for measuring antibody titers in sows. Therefore, the selection of an appropriate ELISA kit according to the purpose of antibody detection is necessary to ensure a more accurate evaluation of the effects of CSFV vaccination on preventing CSF in pig herds.
• Evaluation of the Efficacy of Low-Concentration Gaseous Chlorine Dioxide in Inactivating Airborne H5 High Pathogenicity Avian Influenza Virus in Vivo Model.
  Yik Lim Hew; Norikazu Isoda; Takanori Miura; Takahiro Hiono; Yoshihiro Sakoda
  Food and environmental virology, 18, 1, 4, 4, 23 Jan. 2026, [International Magazine]
  English, Scientific journal, H5 high pathogenicity avian influenza virus (HPAIV) continues to spread globally, causing several high pathogenicity avian influenza (HPAI) outbreaks in poultry and significant economic losses. Biosecurity measures that prevent the introduction of HPAIV represent a top priority for controlling HPAI outbreaks on poultry farms. Although these measures are crucial for minimizing HPAI introduction, outbreaks of viral infection on poultry farms persist, underscoring the importance of continuously improving biosecurity protocols. Therefore, safe and effective microbicide disinfectants could play an essential role in reducing viral spread by inactivating viral particles on surfaces and in the air. This study assessed the efficacy of gaseous chlorine dioxide (ClO2) against H5 HPAIV under both gaseous ClO2 inactivation setting and in vivo conditions. In the gaseous ClO2 inactivation setting, only low virus titers were recovered (< 0.5-1.5 log10 TCID50/mL) when H5 HPAIV aerosols were exposed to gaseous ClO2 (0.05 ppmv, 0.14 mg/m3) for 5 min, corresponding to an approximately 2.0-3.0 log10 reduction. Furthermore, in vivo, all chicks exposed to aerosolized H5 HPAIV, which were treated with 0.1 ppmv gaseous ClO2, survived for 14 days post-challenge, demonstrating complete protection against the virus. The minimum effective concentration of gaseous ClO2 was 0.01 ppmv for 5 min of inactivation in the inactivation setting, and 0.05 ppmv for 5 min in vivo, indicating that relatively low concentrations are sufficient for effective viral inactivation. Therefore, gaseous ClO2 was effective at inactivating aerosolized H5 HPAIV and has potential for use as a disinfectant to prevent HPAIV introduction into poultry. (245/250) words.
• Characterization of H5N1 high pathogenicity avian influenza virus belonging to clade 2.3.4.4b isolated from Ezo red fox in Japan in a mouse model.
  Shintaro Shichinohe; Takahiro Hiono; Yasushi Itoh; Kosuke Takada; Yurie Kida; Pei Wang; Daisuke Motooka; Norikazu Isoda; Ayato Takada; Yoshihiro Sakoda; Tokiko Watanabe
  Microbiology spectrum, 14, 1, e0109725, 06 Jan. 2026, [International Magazine]
  English, Scientific journal, H5N1 high pathogenicity avian influenza virus (HPAIV) has spread in wild birds and poultry worldwide. H5N1 HPAIV belonging to the currently predominant clade 2.3.4.4b has infected not only birds but also mammals (wild and domestic animals), with several human infections also being reported, raising concerns for public health. In 2022, a clade 2.3.4.4b H5N1 HPAIV strain, A/Ezo red fox/Hokkaido/1/2022 (H5N1; Fox/Hok/1/22), was isolated from an Ezo red fox (Vulpes vulpes schrencki) in Hokkaido, Japan; this was the first reported case of clade 2.3.4.4b H5N1 HPAIV isolation from a mammalian species in Japan. Several amino acid substitutions in the PB2 protein play an important role in the adaptation of avian influenza viruses to mammals, but Fox/Hok/1/22 PB2 does not have any of these well-known mammalian-adapting PB2 substitutions. Here, we investigated the biological properties of Fox/Hok/1/22 in a mouse model and found that this virus was highly virulent in mice and replicated well in multiple organs, including the lungs and brain. We then examined whether viruses isolated from these organs acquired known mammalian-adapting PB2 amino acid substitutions, such as PB2 E627K. Deep sequencing analysis of viral RNA from mouse brain and lungs revealed that virus with PB2-627E was predominant in three of four mice, whereas the PB2-627K substitution was predominant in one mouse. These results indicate that Fox/Hok/1/22 is highly virulent in mice despite lacking known PB2 substitutions involved in mammalian adaptation.IMPORTANCEThe H5N1 avian influenza virus has caused severe disease in birds worldwide and is now spreading to mammals, including humans. In 2022, this virus was detected for the first time in an Ezo red fox in Japan. To understand its potential impact on mammals, we studied this virus in mice and found that it caused severe illness, spreading to multiple organs, including the lungs and brain. Surprisingly, despite lacking genetic mutations typically associated with mammalian adaptation, the virus was highly virulent in mice. This finding suggests that the H5N1 virus may pose a greater threat to mammals, including humans, than previously thought. Given their continued spread among wild and domestic animals, our findings underscore the urgent need to monitor how recent H5N1 viruses behave in mammals.
• Introduction and inter-species transmission dynamics of high pathogenicity avian influenza H5N1 viruses in Japan 2021-25.
  Yik Lim Hew; Claire Guinat; Manon Couty; Diletta Fornasiero; Takahiro Hiono; Norikazu Isoda; Yoshihiro Sakoda
  Virus evolution, 12, 1, veag005, 2026, [International Magazine]
  English, Scientific journal, High pathogenicity avian influenza virus impacts poultry and wild birds worldwide. Since the introduction of clade 2.3.4.4b H5N1 isolates in Japan in late 2021, new cases have been reported in domestic birds and poultry each winter. To understand the role of wild birds in introducing these viruses in Japan, a phylodynamic analysis based on geography and host species was conducted using H5N1 isolates in Japan from 2021 to 2025. A total of 892 hemagglutinin gene sequences of H5N1 viruses, collected from birds between June 2021 and May 2025, along with additional sequences that were highly similar to those of Japanese isolates, were obtained from a public database. The role of wild birds in the transmission dynamics of H5N1 isolates in Japan across four winter seasons (2021-25) was assessed using a Bayesian phylodynamic approach with a Multi-Type Birth-Death model. Phylodynamic analysis revealed that the clade 2.3.4.4b comprised three distinct subgroups, G2b, G2c, and G2d, which were prevalent during the winter seasons. Isolates from G2b and G2c were linked to common ancestral strains from North Asia and Northeast Asia, respectively. Meanwhile, G2d, the dominant strain in Japan from 2021 to 2025, shared an ancestral strain from the Northwest America. During winters 2023-25, the ancestral strain was traced back to Northeast Asia, indicating a shift in the viral origin. This transition suggests an increase in virus migration events and expansion of host diversity, implying that Japan may function as a hub for intercontinental virus introductions, receiving multiple independent viral entries from North Asia, Northeast Asia, and Northwest America. Additionally, waterfowl and raptors played a role in introducing viruses into Japan, while poultry and crows generally serving as dead-end hosts. However, the continuous introduction of H5N1 isolates into Japan each winter can alter the disease transmission pattern observed in crows, resulting in more virus spillover from crows to other hosts, such as poultry and charadriiformes. These findings emphasize the importance of continuous monitoring and prompt information sharing to understand the global dynamics of viruses better.
• Risk analysis of infectious disease in pigs in Gifu prefecture, Japan, through network analysis.
  Naotoshi Kuninaga; Emi Yoshita; Miki Koyasu; Hibiki Morimoto; Noboru Hayashi; Takahiro Hiono; Satoshi Ito; Yoshihiro Sakoda; Norikazu Isoda
  Preventive veterinary medicine, 245, 106687, 106687, Dec. 2025, [International Magazine]
  English, Scientific journal, Movements of livestock, humans, vehicles and wildlife are recognized as critical contributors to the spread of infectious diseases in livestock and can be modeled as a network to assess and predict disease transmission. This study developed a comprehensive multilayer network incorporating the movements of pigs, humans, vehicles, and, presumably, wild boars to estimate the risks of disease introduction and transmission for each husbandry stakeholder and to identify key clusters and modes of movement involved. A questionnaire-based study was conducted across 22 pig farms in Gifu Prefecture, Japan, collecting data on pig, human, and vehicle movements to establish networks. The wild boar movement network was estimated using data on pig farm locations and wild boar habitats collected from vegetation cover data. Movement-associated effects in each network based on movement frequency were assigned to combinations of the four networks, resulting in a four-layered network. The network exhibited small-world characteristics and was clustered into four groups. Disease containment schemes in livestock are commonly established along administrative boundaries, however these four epidemiological clusters, comprising 31, 28, 24, and 22 nodes, did not align exactly with administrative districts, suggesting the significance of managing livestock infectious diseases beyond governmental borders. In the Partial Least Squares Regression (PLSR) analysis, pig, vehicle, and wild boar movement made comparable positive contributions to PageRank-based node importance within the multilayer network. This study highlights the significance of epidemiological links among husbandry and nonhusbandry stakeholders, emphasizing the need to develop effective risk management tools considering the probable disease transmission pathways.
• Genetic Diversity of Highly Pathogenic Avian Influenza Viruses Isolated in Hokkaido, Japan, During Winter 2024-2025.
  Norikazu Isoda; Lim Yik Hew; Kazuki Nishikawa; Fumihito Takaya; Yo Shimazu; Daiki Kobayashi; Kei Nabeshima; Hisako Honjyo; Mana Esaki; Kosuke Okuya; Kosuke Soda; Hiroshi Ito; Asuka Kumagai; Hayate Nishiura; Takahiro Hiono; Hiroki Takakuwa; Tatsufumi Usui; Makoto Ozawa; Yuko Uchida; Manabu Onuma; Yoshihiro Sakoda
  Pathogens (Basel, Switzerland), 14, 9, 21 Sep. 2025, [International Magazine]
  English, Scientific journal, Genetic and antigenic analyses were performed on highly pathogenic avian influenza viruses (HPAIVs) isolated in Hokkaido, northern Japan, during the winter of 2024-2025. Ninety-eight HPAIVs were isolated from feces of waterfowl, tracheal swabs from dead wild birds, or lung homogenates from dead chickens. Phylogenetic analysis of the hemagglutinin (HA) gene from 47 representative isolates revealed that all sequences belonged to the G2d subgroup of clade 2.3.4.4b H5HA, which has been the dominant lineage in Hokkaido since the winter of 2021-2022. These isolates were further divided into three major groups within the subgroup. The HPAIVs isolated in the Republic of Korea, China, and North America were genetically closely related to the Hokkaido isolates, whereas no HPAIVs genetically related to European strains or those detected in North American cattle were identified. Furthermore, HPAIVs isolated from seabirds were genetically closely related to those found in dead marine mammals along the eastern coast of Hokkaido in the spring of 2025. No apparent antigenic differences were observed between the HPAIVs isolated in this study and those from previous seasons. These findings highlight the wide distribution of HPAIVs in Hokkaido, particularly from Asian and North American lineages, and underscore the importance of continuous surveillance.
• Dynamics of high pathogenicity avian influenza virus infection with multiple introductions in a crow flock in an urban park in Hokkaido, Japan.
  Norikazu Isoda; Takahiro Hiono; Yik Lim Hew; Fumihito Takaya; Bao Linh Nguyen; Daiki Kobayashi; Kaien Fujino; Yoshihiro Sakoda
  Comparative immunology, microbiology and infectious diseases, 121, 102367, 102367, Aug. 2025, [International Magazine]
  English, Scientific journal, Since 2021, high pathogenicity avian influenza viruses (HPAIVs) of the H5N1 clade 2.3.4.4b has been circulating globally, not only in domestic poultry but also in wild birds, both migratory and resident species. In March to May 2022, March to April 2023, and January to April 2024, crow die-offs were reported in an urban garden in Hokkaido, Japan, raising suspicions of HPAIV infection. Since August 2022, all dead carcasses were investigated for HPAIV detection and isolation. Phylogenetic analysis of the H5 hemagglutinin gene revealed that all detected HPAIVs belonged to clade 2.3.4.4b, a dominant lineage in Hokkaido since early 2022. Two distinct subgroups were identified: G2d (in 2022-2024) and G2a (in 2024). A maximum clade credibility tree, based on concatenated nucleotide sequences of the isolates, suggested that multiple distinct types of HPAIVs were introduced into the garden in rotation during the winters of 2022-2023 and 2023-2024. Infectious HPAIVs were isolated not only from the lungs and brains but also from the rectal contents of the dead crows, with no apparent difference in viral titers between the two subgroups. The case reproduction numbers of HPAIV infection in the crow flock ranged from 0.52 and 1.57 in the spring of 2022 and from 0.55 to 1.78 in the spring of 2023, suggesting that the contiguous HPAIV infections in the crows were due to multiple introductions into the flock. Crow can play a key role of potential spread to other animals, poultry and wildlife in urban areas or humans in rural areas.
• Altered receptor-binding specificity of gull-adapted H13 avian influenza viruses corresponds to their unique host preferences.
  Rio Harada; Takahiro Hiono; Manabu Igarashi; Daiki Kobayashi; Hinako Ban; Norikazu Isoda; Yoshihiro Sakoda
  Virology, 605, 110460, 110460, 21 Feb. 2025, [Corresponding author], [International Magazine]
  English, Scientific journal, Avian influenza viruses (AIVs) recognize α2-3 sialosides as receptors. Previous studies showed that the structural diversity within α2-3 sialosides is related to the host specificity of AIVs. H13 AIVs are primarily isolated from gulls, although almost all AIV subtypes have been isolated from ducks, the natural hosts of AIVs. To elucidate the molecular basis of the host specificity of H13 viruses to gulls, the receptor-binding specificity of H13 hemagglutinins (HAs) and the distribution of viral receptors in gulls were investigated. The results revealed that recombinant HA (rHA) of H13 viruses had a binding preference for fucosylated α2-3 sialosides, which were distributed widely in the respiratory tract and intestines of gulls but not in the colon of ducks. Moreover, the receptor-binding specificity of mutant rHAs revealed that amino acids in the 130-loop and at position 227 of H13 HA were critical for the preference for fucosylated α2-3 sialosides. The results of the present study suggest that the binding specificity of H13 HA to fucosylated α2-3 sialosides is a key factor for the host susceptibility of H13 viruses to gulls.
• Deglycosylation and truncation in the neuraminidase stalk are functionally equivalent in enhancing the pathogenicity of a high pathogenicity avian influenza virus in chickens.
  Daiki Kobayashi; Takahiro Hiono; Hiromu Arakawa; Hiroyuki Kaji; Ayako Ohkawara; Takaya Ichikawa; Hinako Ban; Norikazu Isoda; Yoshihiro Sakoda
  Journal of virology, 99, 3, e0147824, 14 Feb. 2025, [International Magazine]
  English, Scientific journal, Influenza A viruses with fewer amino acids in the neuraminidase (NA) stalk domain are primarily isolated from chickens rather than wild ducks, indicating that a shortened NA stalk is considered an adaptation marker of avian influenza viruses (AIVs) to chickens. Experimental passages of an H7N7 nonpathogenic AIV (rgVac2-P0) in chickens resulted in a highly pathogenic variant (Vac2-P3L4) with a 34-amino-acid deletion in the NA stalk, encompassing five potential N-glycosylation sites. To investigate how amino acid truncation and deglycosylation in the NA stalk contribute to increased pathogenicity, a virus with glycosylation-deficient mutations at these sites (rgVac2-P3L4/P0NAΔGlyco) was constructed. Contrary to expectations, chickens inoculated with rgVac2-P3L4/P0NAΔGlyco exhibited variable clinical outcomes, attributed to the genetic instability of the virus. A single mutation stabilized the virus, and the mutant (rgVac2-P3L4/P0NAΔGlyco-Y65H) resulted in higher pathogenicity compared with a virus with restored glycosylation (rgVac2-P3L4/P0NA-Y65H). Glycan occupancy analysis revealed 3-4 glycans at the five potential sites. In functional analysis, glycosylation-deficient mutants, similar to the short-stalk NA virus, showed significantly reduced erythrocyte elution activity. Additionally, mutational analysis indicated variable contributions of N-glycans to elution activity across the sites. Moreover, the functionally most contributing sites of the five potential N-glycosylation motifs were consistently included in the amino acid deletions of the stalk-truncated NA in N7-subtyped field isolates, despite the varying truncation position or length. These findings suggest that the loss of glycosylation is functionally equivalent to a reduction in amino acids, and it plays a crucial role in enhancing pathogenicity in chickens and affecting NA function.IMPORTANCEAvian influenza poses significant economic challenges to the poultry industry and presents potential risks to human health. Understanding the molecular mechanisms that facilitate the emergence of chicken-adapted avian influenza viruses (AIVs) from non-pathogenic duck-origin influenza viruses is crucial for improving AIV monitoring systems in poultry and controlling this disease. Amino acid deletions in the neuraminidase (NA) stalk domain serve as one of the molecular markers for AIV adaptation to Galliformes. This study highlights the critical role of N-glycosylation in the NA stalk domain in the pathogenesis of high pathogenicity avian influenza viruses in chickens. The findings propose a novel theory that the loss of glycosylation at the NA stalk domain, rather than a reduction in stalk length, is responsible for both NA function and increased virus pathogenicity in chickens.
• Generation of Vaccine Candidate Strains That Antigenically Match Classical Swine Fever Virus Field Strains.
  Maya Kobayashi; Loc Tan Huynh; Saho Ogino; Lim Yik Hew; Miki Koyasu; Hikaru Kamata; Takahiro Hiono; Norikazu Isoda; Yoshihiro Sakoda
  Vaccines, 13, 2, 14 Feb. 2025, [International Magazine]
  English, Scientific journal, BACKGROUND: Classical swine fever virus (CSFV) is genetically categorized into three genotypes. A live-attenuated vaccine strain GPE-, currently used in Japan, belongs to genotype 1 and is genetically distinct from the field strains circulating in Japan, which belong to genotype 2. This study aimed to understand the antigenicity of recent field isolates in Japan and develop new vaccine candidates that antigenically match field strains. METHODS: The serum samples of 20 pigs vaccinated with GPE- were subjected to a serum neutralizing test (SNT) using one of the field strains, CSFV/wb/Jpn-Mie/P96/2019 (Mie/2019). For the antigenic matching, vGPE-/HiBiT/Mie E2 was generated by replacing the viral glycoprotein E2, the main target of the neutralizing antibody, with that of Mie/2019. Additionally, vGPE-/HiBiT/Mie E2/PAPeV Erns was generated by further substituting glycoprotein Erns with that of pronghorn antelope pestivirus (PAPeV) since Erns is not important as a vaccine immunogen and can be replaced by that of other pestiviruses to provide an immunological marker. The efficacy of vGPE-/HiBiT/Mie E2/PAPeV Erns was further evaluated by the challenge experiments in pigs. RESULTS: The SNT titers of serum sample against Mie/2019 were 6.1-fold lower than that against vGPE-. The generated recombinant viruses showed closer antigenicity to Mie/2019 than vGPE-. The challenge study confirmed that vGPE-/HiBiT/Mie E2/PAPeV Erns provided clinical and virological protection against a field CSFV equivalent to vGPE-. CONCLUSIONS: This study demonstrated that swapping the E2 encoding region with the prevalent field CSFVs is a promising strategy to achieve antigenic matching between the vaccine and field strains.
• Serosurvey of Bovine Viral Diarrhea Virus in Cattle in Southern Japan and Estimation of Its Transmissibility by Transient Infection in Nonvaccinated Cattle.
  Norikazu Isoda; Satoshi Sekiguchi; Chika Ryu; Kosuke Notsu; Maya Kobayashi; Karin Hamaguchi; Takahiro Hiono; Yuichi Ushitani; Yoshihiro Sakoda
  Viruses, 17, 1, 02 Jan. 2025, [International Magazine]
  English, Scientific journal, Bovine viral diarrhea (BVD) is caused by the BVD virus (BVDV) and has been reported worldwide in cattle. To estimate BVDV circulation among cattle where few BVD cases were reported in southern Japan, 1910 serum samples collected from 35 cattle farms without a BVD outbreak were investigated to detect antibodies against BVDV-1 and BVDV-2 using an indicator virus with a cytopathogenic effect and the luciferase gene, respectively. Neutralizing antibodies against BVDV-1 and BVDV-2 were detected more frequently in 18 vaccinated farms than in 17 nonvaccinated farms. In the nonvaccinated farms, 9.6%, 1.8%, and 13.8% of the cattle were estimated to have a history of infection with BVDV-1, BVDV-2, and both, respectively. The median rate of within-herd anti-BVDV-1 seropositivity among cattle in the nonvaccinated farms was 22.0%; however, a high within-herd seropositivity (>50%) was confirmed in the two farms. The force of infection, basic reproduction number, and annual probability of BVDV-1 infection were estimated as 0.072 (95% confidence interval [CI]: 0.062-0.084), 0.36 (95% CI: 0.31-0.42), and 0.73% (95% CI: 0.61-0.87%), respectively, using the age-specific positive rate of anti-BVDV-1 antibodies. These parameters should be further applicable for developing epidemiological models which illustrate the BVDV dynamics in the field.
• Hemagglutinin and neuraminidase of a non-pathogenic H7N7 avian influenza virus coevolved during the acquisition of intranasal pathogenicity in chickens
  Takaya Ichikawa; Takahiro Hiono; Masatoshi Okamatsu; Junki Maruyama; Daiki Kobayashi; Keita Matsuno; Hiroshi Kida; Yoshihiro Sakoda
  Archives of Virology, 169, 10, 207, 207, Oct. 2024, [International Magazine]
  English, Scientific journal, Polybasic amino acid residues at the hemagglutinin (HA) cleavage site are insufficient to induce the highly pathogenic phenotype of avian influenza viruses in chickens. In our previous study, an H7N7 avian influenza virus named "Vac2sub-P0", which is nonpathogenic despite carrying polybasic amino acids at the HA cleavage site, was passaged in chick air sacs, and a virus with high intravenous pathogenicity, Vac2sub-P3, was obtained. Intranasal infection with Vac2sub-P3 resulted in limited lethality in chickens; therefore, in this study, this virus was further passaged in chicken lungs, and the resultant virus, Vac2sub-P3L4, acquired high intranasal pathogenicity. Experimental infection of chickens with recombinant viruses demonstrated that mutations in HA and neuraminidase (NA) found in consecutive passages were responsible for the increased pathogenicity. The HA and NA functions of Vac2sub-P3L4 were compared with those of the parental virus in vitro; the virus growth at 40 °C was faster, the binding affinity to a sialic acid receptor was lower, and the rate of release by NA from the cell surface was lower, suggesting that these changes enabled the virus to replicate efficiently in chickens with high intranasal pathogenicity. This study demonstrates that viruses that are highly pathogenic when administered intranasally require additional adaptations for increased pathogenicity to be highly lethal to intranasally infected chickens.
• Cocirculation of Genetically Distinct Highly Pathogenic Avian Influenza H5N5 and H5N1 Viruses in Crows, Hokkaido, Japan.
  Yik Lim Hew; Takahiro Hiono; Isabella Monne; Kei Nabeshima; Saki Sakuma; Asuka Kumagai; Shunya Okamura; Kosuke Soda; Hiroshi Ito; Mana Esaki; Kosuke Okuya; Makoto Ozawa; Toshiyo Yabuta; Hiroki Takakuwa; Linh Bao Nguyen; Norikazu Isoda; Kohtaro Miyazawa; Manabu Onuma; Yoshihiro Sakoda
  Emerging infectious diseases, 30, 9, 1912, 1917, 06 Aug. 2024, [Corresponding author], [International Magazine]
  English, Scientific journal, We isolated highly pathogenic avian influenza (HPAI) H5N5 and H5N1 viruses from crows in Hokkaido, Japan, during winter 2023-24. They shared genetic similarity with HPAI H5N5 viruses from northern Europe but differed from those in Asia. Continuous monitoring and rapid information sharing between countries are needed to prevent HPAI virus transmission.
• Assessment of the Safety Profile of Chimeric Marker Vaccine against Classical Swine Fever: Reversion to Virulence Study.
  Loc Tan Huynh; Mikihiro Otsuka; Maya Kobayashi; Hung Dinh Ngo; Lim Yik Hew; Takahiro Hiono; Norikazu Isoda; Yoshihiro Sakoda
  Viruses, 16, 7, 12 Jul. 2024, [International Magazine]
  English, Scientific journal, Chimeric marker vaccine candidates, vGPE-/PAPeV Erns and vGPE-/PhoPeV Erns, have been generated and their efficacy and capability to differentiate infected from vaccinated animals were confirmed in previous studies. The safety profile of the two chimeric marker vaccine candidates, particularly in the potential reversion to virulence, was evaluated. Each virus was administered to pigs with a dose equivalent to the vaccination dose, and pooled tonsil homogenates were subsequently inoculated into further pigs. Chimeric virus vGPE-/PAPeV Erns displayed the most substantial attenuation, achieving this within only two passages, whereas vGPE-/PhoPeV Erns was detectable until the third passage and disappeared entirely by the fourth passage. The vGPE- strain, assessed alongside, consistently exhibited stable virus recovery across each passage without any signs of increased virulence in pigs. In vitro assays revealed that the type I interferon-inducing capacity of vGPE-/PAPeV Erns was significantly higher than that of vGPE-/PhoPeV Erns and vGPE-. In conclusion, the safety profile of the two chimeric marker vaccine candidates was affirmed. Further research is essential to ensure the stability of their attenuation and safety in diverse pig populations.
• FOUR-WEEK ORAL ADMINISTRATION OF BALOXAVIR MARBOXIL AS AN ANTI-INFLUENZA VIRUS DRUG SHOWS NO TOXICITY IN CHICKENS.
  Mariko Miki; Ryo Daniel Obara; Kyohei Nishimura; Takao Shishido; Yoshinori Ikenaka; Ryoko Oka; Kenji Sato; Shouta M M Nakayama; Takashi Kimura; Atsushi Kobayashi; Keisuke Aoshima; Keisuke Saito; Takahiro Hiono; Norikazu Isoda; Yoshihiro Sakoda
  Journal of zoo and wildlife medicine : official publication of the American Association of Zoo Veterinarians, 55, 2, 313, 321, Jun. 2024, [International Magazine]
  English, Scientific journal, High pathogenicity avian influenza is an acute zoonotic disease with high mortality in birds caused by a high pathogenicity avian influenza virus (HPAIV). Recently, HPAIV has rapidly spread worldwide and has killed many wild birds, including endangered species. Baloxavir marboxil (BXM), an anti-influenza agent used for humans, was reported to reduce mortality and virus secretion from HPAIV-infected chickens (Gallus domesticus, order Galliformes) at a dosage of ≥2.5 mg/kg when administered simultaneously with viral challenge. Application of this treatment to endangered birds requires further information on potential avian-specific toxicity caused by repeated exposure to BXM over the long term. To obtain information of potential avian-specific toxicity, a 4-wk oral repeated-dose study of BXM was conducted in chickens (n = 6 or 7 per group), which are commonly used as laboratory avian species. The study was conducted in reference to the human pharmaceutical guidelines for nonclinical repeated-dose drug toxicity studies to evaluate systemic toxicity and exposure. No adverse changes were observed in any organs examined, and dose proportional increases in systemic exposure to active pharmaceutical ingredients were noted from 12.5 to 62.5 mg/kg per day. BXM showed no toxicity to chickens at doses of up to 62.5 mg/kg per day, at which systemic exposure was approximately 71 times higher than systemic exposure at 2.5 mg/kg, the reported efficacious dosage amount, in HPAIV-infected chickens. These results also suggest that BXM could be considered safe for treating HPAIV-infected endangered birds due to its high safety margin compared with the efficacy dose. The data in this study could contribute to the preservation of endangered birds by using BXM as a means of protecting biodiversity.
• Development of a dual immunochromatographic test strip to detect E2 and Erns antibodies against classical swine fever
  Loc Tan Huynh; Eun-Ju Sohn; Youngmin Park; Juhun Kim; Tomohiko Shimoda; Takahiro Hiono; Norikazu Isoda; Sung-Hee Hong; Ha-Na Lee; Yoshihiro Sakoda
  Frontiers in Microbiology, 15, 1383976, 1383976, Frontiers Media SA, 11 Apr. 2024, [International Magazine]
  English, Scientific journal, Background
  
  It is essential to consider a practical antibody test to successfully implement marker vaccines and validate vaccination efficacy against classical swine fever virus (CSFV). The test should include a serological antibody assay, combined with a tool for differentiating infected from vaccinated animals (DIVA). The immunochromatographic test strip (ICS) has been exclusively designed for detecting CSFV E2 antibodies while lacking in detecting E^rns antibodies, which can be employed and satisfy DIVA strategy. This study developed a novel ICS for detecting CSFV E2/E^rns dual-antibody. The effectiveness of ICS in evaluating the DIVA capability of two novel chimeric pestivirus vaccine candidates was assessed.
  
  Methods
  
  Recombinant E2 or E^rns protein was transiently expressed in the plant benthamiana using Agrobacterium tumefaciens. ICS was subsequently assembled, and goat anti-rabbit IgG and recombinant CSFV E2 or E^rns protein were plated onto the nitrocellulose membrane as control and test lines, respectively. The sensitivity and specificity of ICS were evaluated using sera with different neutralizing antibody titers or positive for antibodies against CSFV and other pestiviruses. The coincidence rates for detecting E2 and E^rns antibodies between ICS and commercial enzyme-linked immunosorbent assay (ELISA) kits were also computed. ICS performance for DIVA capability was evaluated using sera from pigs vaccinated with conventional vaccine or chimeric vaccine candidates.
  
  Results
  
  E2 and E^rns proteins were successfully expressed in N. benthamiana-produced recombinant proteins. ICS demonstrated high sensitivity in identifying CSFV E2 and E^rns antibodies, even at the low neutralizing antibody titers. No cross-reactivity with antibodies from other pestiviruses was confirmed using ICS. There were high agreement rates of 93.0 and 96.5% between ICS and two commercial ELISA kits for E2 antibody testing. ICS also achieved strong coincidence rates of 92.9 and 89.3% with two ELISA kits for E^rns antibody detection. ICS confirmed the absence of CSFV E^rns-specific antibodies in sera from pigs vaccinated with chimeric vaccine candidates.
  
  Conclusion
  
  E2 and E^rns proteins derived from the plant showed great potential and can be used to engineer a CSFV E2/E^rns dual-antibody ICS. The ICS was also highly sensitive and specific for detecting CSFV E2 and E^rns antibodies. Significantly, ICS can fulfill the DIVA concept by incorporating chimeric vaccine candidates.
• Combinatorial Approach with Mass Spectrometry and Lectin Microarray Dissected Site-Specific Glycostem and Glycoleaf Features of the Virion-Derived Spike Protein of Ancestral and γ Variant SARS-CoV-2 Strains.
  Takahiro Hiono; Hiroaki Sakaue; Azusa Tomioka; Hiroyuki Kaji; Michihito Sasaki; Yasuko Orba; Hirofumi Sawa; Atsushi Kuno
  Journal of proteome research, 23, 4, 1408, 1419, 05 Apr. 2024, [Lead author, Corresponding author], [International Magazine]
  English, Scientific journal, The coronavirus disease (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has impacted public health globally. As the glycosylation of viral envelope glycoproteins is strongly associated with their immunogenicity, intensive studies have been conducted on the glycans of the glycoprotein of SARS-CoV-2, the spike (S) protein. Here, we conducted intensive glycoproteomic analyses of the SARS-CoV-2 S protein of ancestral and γ-variant strains using a combinatorial approach with two different technologies: mass spectrometry (MS) and lectin microarrays (LMA). Our unique MS1-based glycoproteomic technique, Glyco-RIDGE, in addition to MS2-based Byonic search, identified 1448 (ancestral strain) and 1785 (γ-variant strain) site-specific glycan compositions, respectively. Asparagine at amino acid position 20 (N20) is mainly glycosylated within two successive potential glycosylation sites, N17 and N20, of the γ-variant S protein; however, we found low-frequency glycosylation at N17. Our novel approaches, glycostem mapping and glycoleaf scoring, also illustrate the moderately branched/extended, highly fucosylated, and less sialylated natures of the glycoforms of S proteins. Subsequent LMA analysis emphasized the intensive end-capping of glycans by Lewis fucoses, which complemented the glycoproteomic features. These results illustrate the high-resolution glycoproteomic features of the SARS-CoV-2 S protein, contributing to vaccine design and understanding of viral protein synthesis.
• Continuous Introduction of H5 High Pathogenicity Avian Influenza Viruses in Hokkaido, Japan: Characterization of Viruses Isolated in Winter 2022–2023 and Early Winter 2023–2024
  Lim Yik Hew; Norikazu Isoda; Fumihito Takaya; Kohei Ogasawara; Daiki Kobayashi; Loc Tan Huynh; Tatsuru Morita; Rio Harada; Nikolay Gennadievich Zinyakov; Dmitriy Borisovich Andreychuk; Ilya Alexandrovich Chvala; Viktor Nikolaevich Irza; Yukiko Watanabe; Hiroko Fujita; Keisuke Saito; Takahiro Hiono; Yoshihiro Sakoda
  Transboundary and Emerging Diseases, 2024, 1, 18, Hindawi Limited, 14 Mar. 2024, [International Magazine]
  English, Scientific journal, High pathogenicity avian influenza (HPAI) has impacted poultry and wild birds globally. The number of H5 HPAI virus (HPAIV) infection cases in wild birds in Hokkaido (Northern Japan) was high in the last two seasons, contributing to virus spillover to resident birds and poultry. Therefore, H5 HPAIVs in birds and mammals in Hokkaido in winter 2022–2023 and 2023–2024 were monitored and viruses were phylogenetically, antigenically, and pathogenetically characterized. Thirty HPAIV isolates were subtyped and pathotyped by sequencing the hemagglutinin (HA) gene of viruses. Phylogenetic analysis of the HA gene revealed that all isolated HPAIVs were categorized into clade 2.3.4.4b and divided into three groups (G2b, G2c, and G2d). Most isolates belonging to subgroup G2d clustered with isolates in winter 2021–2022 in Hokkaido. The other isolates were categorized into two subgroups, G2b and G2c, mainly composed of isolates in Honshu Island in winter 2021–2022 and 2022–2023, respectively. Two H5 HPAIVs isolated in Eastern Russia in spring and autumn 2022 were genetically close to most Hokkaido isolates (G2d), and a virus isolated in Hokkaido in November 2023 was also grouped in subgroup G2d. Further analysis of all eight gene segments identified six types of gene constellations. Cross-hemagglutination inhibition test indicated that the antigenicity of H5 HPAIVs isolated in the last several seasons was similar within them but slightly different from that in the 2010s. Three chicken breeds were intranasally challenged with four representative isolates to assess their pathogenicity. All chickens except one broiler chicken were dead until 5-day postchallenge with different pathogenicity of these viruses. The pathogenicity of one HPAIV strain was significantly lower in broiler chickens than in layer chickens. The mixture of multiple characteristics of HPAIVs in Hokkaido was confirmed by bird migration routes. Thus, many HPAIVs can be brought and scattered anywhere on Earth.
• A rapid and versatile reverse genetics approach for generating recombinant positive-strand RNA viruses that use IRES-mediated translation.
  Tomokazu Tamura; Hirotaka Yamamoto; Saho Ogino; Yuhei Morioka; Shuhei Tsujino; Rigel Suzuki; Takahiro Hiono; Saori Suzuki; Norikazu Isoda; Yoshihiro Sakoda; Takasuke Fukuhara
  Journal of virology, e0163823, 14 Feb. 2024, [International Magazine]
  English, Scientific journal, Reverse genetics systems have played a central role in developing recombinant viruses for a wide spectrum of virus research. The circular polymerase extension reaction (CPER) method has been applied to studying positive-strand RNA viruses, allowing researchers to bypass molecular cloning of viral cDNA clones and thus leading to the rapid generation of recombinant viruses. However, thus far, the CPER protocol has only been established using cap-dependent RNA viruses. Here, we demonstrate that a modified version of the CPER method can be successfully applied to positive-strand RNA viruses that use cap-independent, internal ribosomal entry site (IRES)-mediated translation. As a proof-of-concept, we employed mammalian viruses with different types (classes I, II, and III) of IRES to optimize the CPER method. Using the hepatitis C virus (HCV, class III), we found that inclusion in the CPER assembly of an RNA polymerase I promoter and terminator, instead of those from polymerase II, allowed greater viral production. This approach was also successful in generating recombinant bovine viral diarrhea virus (class III) following transfection of MDBK/293T co-cultures to overcome low transfection efficiency. In addition, we successfully generated the recombinant viruses from clinical specimens. Our modified CPER could be used for producing hepatitis A virus (HAV, type I) as well as de novo generation of encephalomyocarditis virus (type II). Finally, we generated recombinant HCV and HAV reporter viruses that exhibited replication comparable to that of the wild-type parental viruses. The recombinant HAV reporter virus helped evaluate antivirals. Taking the findings together, this study offers methodological advances in virology.IMPORTANCEThe lack of versatility of reverse genetics systems remains a bottleneck in viral research. Especially when (re-)emerging viruses reach pandemic levels, rapid characterization and establishment of effective countermeasures using recombinant viruses are beneficial in disease control. Indeed, numerous studies have attempted to establish and improve the methods. The circular polymerase extension reaction (CPER) method has overcome major obstacles in generating recombinant viruses. However, this method has not yet been examined for positive-strand RNA viruses that use cap-independent, internal ribosome entry site-mediated translation. Here, we engineered a suitable gene cassette to expand the CPER method for all positive-strand RNA viruses. Furthermore, we overcame the difficulty of generating recombinant viruses because of low transfection efficiency. Using this modified method, we also successfully generated reporter viruses and recombinant viruses from a field sample without virus isolation. Taking these findings together, our adapted methodology is an innovative technology that could help advance virologic research.
• Establishment of a superinfection exclusion method for pestivirus titration using a recombinant reporter pestiviruses
  Yume MIMURA; Takahiro HIONO; Loc Tan HUYNH; Saho OGINO; Maya KOBAYASHI; Norikazu ISODA; Yoshihiro SAKODA
  Journal of Veterinary Medical Science, 86, 4, 389, 395, Japanese Society of Veterinary Science, 2024, [Domestic magazines]
  English, Scientific journal, Pestiviruses are classified into two biotypes based on their cytopathogenicity. As the majority of pestivirus field isolates are noncytopathogenic, their titration requires alternative methods rather than direct observation of cytopathogenic effects, such as immunostaining using specific antibodies or interference with cytopathogenic strains. However, these methods require microscopic observation to assess virus growth, which is time- and labor-intensive, especially when handling several samples. In this study, we developed a novel luciferase-based pestivirus titration method using the superinfection exclusion phenomenon with recombinant reporter pestiviruses that possessed an 11-amino-acid subunit derived from NanoLuc luciferase (HiBiT). In this method, swine kidney cells were inoculated with classical swine fever virus (CSFV) and superinfected with the reporter CSFV vGPE-/HiBiT 5 days postinoculation. Virus titer was determined based on virus growth measured in luminescence using the culture fluid 3 days after superinfection; the resultant virus titer was comparable to that obtained by immunoperoxidase staining. Furthermore, this method has proven to be applicable for the titration of border disease virus (BDV) by superinfection with both the homologous reporter BDV and heterologous reporter CSFV, suggesting that this novel virus titration method is a simple technique for automated virus detection based on the luciferase system.
• Detection of H5N1 High Pathogenicity Avian Influenza Viruses in Four Raptors and Two Geese in Japan in the Fall of 2022.
  Kei Nabeshima; Yoshihiro Takadate; Kosuke Soda; Takahiro Hiono; Norikazu Isoda; Yoshihiro Sakoda; Junki Mine; Kohtaro Miyazawa; Manabu Onuma; Yuko Uchida
  Viruses, 15, 9, 01 Sep. 2023, [International Magazine]
  English, Scientific journal, In the fall of 2022, high pathogenicity avian influenza viruses (HPAIVs) were detected from raptors and geese in Japan, a month earlier than in past years, indicating a shift in detection patterns. In this study, we conducted a phylogenetic analysis on H5N1 HPAIVs detected from six wild birds during the 2022/2023 season to determine their genetic origins. Our findings revealed that these HPAIVs belong to the G2 group within clade 2.3.4.4b, with all isolates classified into three subgroups: G2b, G2d, and G2c. The genetic background of the G2b virus (a peregrine falcon-derived strain) and G2d viruses (two raptors and two geese-derived strains) were the same as those detected in Japan in the 2021/2022 season. Since no HPAI cases were reported in Japan during the summer of 2022, it is probable that migratory birds reintroduced the G2b and G2d viruses. Conversely, the G2c virus (a raptor-derived strain) was first recognized in Japan in the fall of 2022. This strain might share a common ancestor with HPAIVs from Asia and West Siberia observed in the 2021/2022 season. The early migration of waterfowl to Japan in the fall of 2022 could have facilitated the early invasion of HPAIVs.
• Generation and Efficacy of Two Chimeric Viruses Derived from GPE− Vaccine Strain as Classical Swine Fever Vaccine Candidates
  Loc Tan Huynh; Norikazu Isoda; Lim Yik Hew; Saho Ogino; Yume Mimura; Maya Kobayashi; Taksoo Kim; Tatsuya Nishi; Katsuhiko Fukai; Takahiro Hiono; Yoshihiro Sakoda
  Viruses, 15, 7, 1587, 1587, MDPI AG, 20 Jul. 2023
  Scientific journal, A previous study proved that vGPE− mainly maintains the properties of classical swine fever (CSF) virus, which is comparable to the GPE− vaccine seed and is a potentially valuable backbone for developing a CSF marker vaccine. Chimeric viruses were constructed based on an infectious cDNA clone derived from the live attenuated GPE− vaccine strain as novel CSF vaccine candidates that potentially meet the concept of differentiating infected from vaccinated animals (DIVA) by substituting the glycoprotein Erns of the GPE− vaccine strain with the corresponding region of non-CSF pestiviruses, either pronghorn antelope pestivirus (PAPeV) or Phocoena pestivirus (PhoPeV). High viral growth and genetic stability after serial passages of the chimeric viruses, namely vGPE−/PAPeV Erns and vGPE−/PhoPeV Erns, were confirmed in vitro. In vivo investigation revealed that two chimeric viruses had comparable immunogenicity and safety profiles to the vGPE− vaccine strain. Vaccination at a dose of 104.0 TCID50 with either vGPE−/PAPeV Erns or vGPE−/PhoPeV Erns conferred complete protection for pigs against the CSF virus challenge in the early stage of immunization. In conclusion, the characteristics of vGPE−/PAPeV Erns and vGPE−/PhoPeV Erns affirmed their properties, as the vGPE− vaccine strain, positioning them as ideal candidates for future development of a CSF marker vaccine.
• Inhibition of cellular RNA methyltransferase abrogates influenza virus capping and replication.
  Yuta Tsukamoto; Takahiro Hiono; Shintaro Yamada; Keita Matsuno; Aileen Faist; Tobias Claff; Jianyu Hou; Vigneshwaran Namasivayam; Anja Vom Hemdt; Satoko Sugimoto; Jin Ying Ng; Maria H Christensen; Yonas M Tesfamariam; Steven Wolter; Stefan Juranek; Thomas Zillinger; Stefan Bauer; Takatsugu Hirokawa; Florian I Schmidt; Georg Kochs; Masayuki Shimojima; Yi-Shuian Huang; Andreas Pichlmair; Beate M Kümmerer; Yoshihiro Sakoda; Martin Schlee; Linda Brunotte; Christa E Müller; Manabu Igarashi; Hiroki Kato
  Science (New York, N.Y.), 379, 6632, 586, 591, 10 Feb. 2023, [International Magazine]
  English, Scientific journal, Orthomyxo- and bunyaviruses steal the 5' cap portion of host RNAs to prime their own transcription in a process called "cap snatching." We report that RNA modification of the cap portion by host 2'-O-ribose methyltransferase 1 (MTr1) is essential for the initiation of influenza A and B virus replication, but not for other cap-snatching viruses. We identified with in silico compound screening and functional analysis a derivative of a natural product from Streptomyces, called trifluoromethyl-tubercidin (TFMT), that inhibits MTr1 through interaction at its S-adenosyl-l-methionine binding pocket to restrict influenza virus replication. Mechanistically, TFMT impairs the association of host cap RNAs with the viral polymerase basic protein 2 subunit in human lung explants and in vivo in mice. TFMT acts synergistically with approved anti-influenza drugs.
• Genetic, Antigenic, and Pathobiological Characterization of H9 and H6 Low Pathogenicity Avian Influenza Viruses Isolated in Vietnam from 2014 to 2018.
  Kien Trung Le; Lam Thanh Nguyen; Loc Tan Huynh; Duc-Huy Chu; Long Van Nguyen; Tien Ngoc Nguyen; Tien Ngoc Tien; Keita Matsuno; Masatoshi Okamatsu; Takahiro Hiono; Norikazu Isoda; Yoshihiro Sakoda
  Microorganisms, 11, 2, 18 Jan. 2023, [International Magazine]
  English, Scientific journal, The H9 and H6 subtypes of low pathogenicity avian influenza viruses (LPAIVs) cause substantial economic losses in poultry worldwide, including Vietnam. Herein, we characterized Vietnamese H9 and H6 LPAIVs to facilitate the control of avian influenza. The space-time representative viruses of each subtype were selected based on active surveillance from 2014 to 2018 in Vietnam. Phylogenetic analysis using hemagglutinin genes revealed that 54 H9 and 48 H6 Vietnamese LPAIVs were classified into the sublineages Y280/BJ94 and Group II, respectively. Gene constellation analysis indicated that 6 and 19 genotypes of the H9 and H6 subtypes, respectively, belonged to the representative viruses. The Vietnamese viruses are genetically related to the previous isolates and those in neighboring countries, indicating their circulation in poultry after being introduced into Vietnam. The antigenicity of these subtypes was different from that of viruses isolated from wild birds. Antigenicity was more conserved in the H9 viruses than in the H6 viruses. Furthermore, a representative H9 LPAIV exhibited systemic replication in chickens, which was enhanced by coinfection with avian pathogenic Escherichia coli O2. Although H9 and H6 were classified as LPAIVs, their characterization indicated that their silent spread might significantly affect the poultry industry.
• Virological, pathological, and glycovirological investigations of an Ezo red fox and a tanuki naturally infected with H5N1 highly pathogenic avian influenza viruses in Hokkaido, Japan
  Takahiro Hiono; Daiki Kobayashi; Atsushi Kobayashi; Tamami Suzuki; Yuki Satake; Rio Harada; Keita Matsuno; Mariko Sashika; Hinako Ban; Maya Kobayashi; Keisuke Aoshima; Fumihito Takaya; Hiroko Fujita; Norikazu Isoda; Takashi Kimura; Yoshihiro Sakoda
  Virology, 578, 35, 44, Jan. 2023, [Peer-reviewed], [Lead author], [International Magazine]
  English, Scientific journal, In winter/spring 2021-2022, high pathogenicity avian influenza viruses (HPAIVs) that are genetically closely related to each other were detected worldwide. In a public garden in Sapporo, Hokkaido, Japan, a crow die-off by HPAIV infection occurred from March 29 to May 18, 2022. During the event, H5N1 HPAIVs were isolated from an Ezo red fox (Vulpes vulpes schrencki) and a tanuki (Nyctereutes procyonoides albus) found in the same garden. The fox showed viral meningoencephalitis and moderate virus replication in the upper respiratory tract, whereas the tanuki showed viral conjunctivitis and secondary bacterial infection in the eyes accompanied with visceral larva migrans. Viruses isolated from the fox and the tanuki were genetically closely related to those isolated from crows in the same garden. Various α2-3 sialosides were found in the respiratory tracts of these canid mammals, consistent with HPAIV infections in these animals. This study highlighted the importance of monitoring HPAIV infections in wild carnivore mammals to detect the potential virus spreading in nature.
• Detection of New H5N1 High Pathogenicity Avian Influenza Viruses in Winter 2021-2022 in the Far East, Which Are Genetically Close to Those in Europe.
  Norikazu Isoda; Manabu Onuma; Takahiro Hiono; Ivan Sobolev; Hew Yik Lim; Kei Nabeshima; Hisako Honjyo; Misako Yokoyama; Alexander Shestopalov; Yoshihiro Sakoda
  Viruses, 14, 10, 30 Sep. 2022, [International Magazine]
  English, Scientific journal, Many high pathogenicity avian influenza (HPAI) cases in wild birds due to H5N1 HPAI virus (HPAIV) infection were reported in northern Japan in the winter of 2021-2022. To investigate the epidemiology of HPAIVs brought to Japan from surrounding areas, a genetic analysis of H5 HPAIVs isolated in northern Japan was performed, and the pathogenicity of the HPAIV in chickens was assessed by experimental infection. Based on the genetic analysis of the hemagglutinin gene, pathogenic viruses detected in northern Japan as well as one in Sakhalin, the eastern part of Russia, were classified into the same subgroup as viruses prevalent in Europe in the same season but distinct from those circulating in Asia in winter 2020-2021. High identities of all eight segment sequences of A/crow/Hokkaido/0103B065/2022 (H5N1) (Crow/Hok), the representative isolates in northern Japan in 2022, to European isolates in the same season could also certify the unlikeliness of causing gene reassortment between H5 HPAIVs and viruses locally circulating in Asia. According to intranasal challenge results in six-week-old chickens, 50% of the chicken-lethal dose of Crow/Hok was calculated as 104.5 times of the 50% egg-infectious dose. These results demonstrated that the currently prevalent H5 HPAIVs could spread widely from certain origins throughout the Eurasian continent, including Europe and the Far East, and implied a possibility that contagious viruses are gathered in lakes in the northern territory via bird migration. Active monitoring of wild birds at the global level is essential to estimate the geographical source and spread dynamics of HPAIVs.
• Influenza A Virus Agnostic Receptor Tropism Revealed Using a Novel Biological System with Terminal Sialic Acid Knockout Cells.
  Haruhiko Kamiki; Shin Murakami; Takashi Nishikaze; Takahiro Hiono; Manabu Igarashi; Yuki Furuse; Hiromichi Matsugo; Hiroho Ishida; Misa Katayama; Wataru Sekine; Yasushi Muraki; Masateru Takahashi; Akiko Takenaka-Uema; Taisuke Horimoto
  Journal of virology, e0041622, 18 Jul. 2022, [International Magazine]
  English, Scientific journal, Avian or human influenza A viruses bind preferentially to avian- or human-type sialic acid receptors, respectively, indicating that receptor tropism is an important factor for determining the viral host range. However, there are currently no reliable methods for analyzing receptor tropism biologically under physiological conditions. In this study, we established a novel system using MDCK cells with avian- or human-type sialic acid receptors and with both sialic acid receptors knocked out (KO). When we examined the replication of human and avian influenza viruses in these KO cells, we observed unique viral receptor tropism that could not be detected using a conventional solid-phase sialylglycan binding assay, which directly assesses physical binding between the virus and sialic acids. Furthermore, we serially passaged an engineered avian-derived H4N5 influenza virus, whose PB2 gene was deleted, in avian-type receptor KO cells stably expressing PB2 to select a mutant with enhanced replication in KO cells; however, its binding to human-type sialylglycan was undetectable using the solid-phase binding assay. These data indicate that a panel of sialic acid receptor KO cells could be a useful tool for determining the biological receptor tropism of influenza A viruses. Moreover, the PB2KO virus experimental system could help to safely and efficiently identify the mutations required for avian influenza viruses to adapt to human cells that could trigger a new influenza pandemic. IMPORTANCE The acquisition of mutations that allow avian influenza A virus hemagglutinins to recognize human-type receptors is mandatory for the transmission of avian viruses to humans, which could lead to a pandemic. In this study, we established a novel system using a set of genetically engineered MDCK cells with knocked out sialic acid receptors to biologically evaluate the receptor tropism for influenza A viruses. Using this system, we observed unique receptor tropism in several virus strains that was undetectable using conventional solid-phase binding assays that measure physical binding between the virus and artificially synthesized sialylglycans. This study contributes to elucidation of the relationship between the physical binding of virus and receptor and viral infectivity. Furthermore, the system using sialic acid knockout cells could provide a useful tool to explore the sialic acid-independent entry mechanism. In addition, our system could be safely used to identify mutations that could acquire human-type receptor tropism.
• Turkeys possess diverse Siaα2-3Gal glycans that facilitate their dual susceptibility to avian influenza viruses isolated from ducks and chickens
  Daiki Kobayashi; Takahiro Hiono; Osamu Ichii; Shoko Nishihara; Sayaka Takase-Yoden; Kazuo Yamamoto; Hiroto Kawashima; Norikazu Isoda; Yoshihiro Sakoda
  Virus Research, 315, 02 Jul. 2022, [Peer-reviewed]
  English, Scientific journal
• Risk profile of low pathogenicity avian influenza virus infections in farms in southern Vietnam.
  Kien Trung LE; Norikazu Isoda; Lam Thanh Nguyen; Duc-Huy Chu; Long Van Nguyen; Minh Quang Phan; Diep Thi Nguyen; Tien Ngoc Nguyen; Tien Ngoc Tien; Tung Thanh LE; Takahiro Hiono; Keita Matsuno; Masatoshi Okamatsu; Yoshihiro Sakoda
  The Journal of veterinary medical science, 84, 6, 860, 868, 13 May 2022, [Domestic magazines]
  English, Scientific journal, The impact of low pathogenicity avian influenza (LPAI) has been confirmed mainly in farms. Unlike apparent losses caused by the high pathogenicity avian influenza (HPAI), the LPAI impact has been hardly evaluated due to underestimating its spread and damage. In 2019, a questionnaire study was conducted in southern Vietnam to identify the specific risk factors of LPAI virus (LPAIV) circulation and to find associations between husbandry activities and LPAI prevalence. A multilevel regression analysis indicated that keeping Muscovy ducks during farming contributed to LPAIV positivity [Odds ratio=208.2 (95% confidence interval: 13.4-1.1×104)]. In cluster analysis, farmers willing to report avian influenza (AI) events and who agreed with the local AI control policy had a slightly lower risk for LPAIV infection although there was no significance in the correlation between farmer characteristics and LPAI occurrence. These findings indicated that keeping Muscovy ducks without appropriate countermeasures might increase the risk of LPAIV infection. Furthermore, specific control measures at the local level are effective for LPAIV circulation, and the improvement of knowledge about biosecurity and attitude contributes to reducing LPAI damage.
• Susceptibility of herons (family: Ardeidae) to clade 2.3.2.1 H5N1 subtype high pathogenicity avian influenza virus.
  Kosuke Soda; Yukiko Tomioka; Tatsufumi Usui; Yukiko Uno; Yasuko Nagai; Hiroshi Ito; Takahiro Hiono; Tomokazu Tamura; Masatoshi Okamatsu; Masahiro Kajihara; Naganori Nao; Yoshihiro Sakoda; Ayato Takada; Toshihiro Ito
  Avian pathology : journal of the W.V.P.A, 51, 2, 146, 153, Apr. 2022, [International Magazine]
  English, Scientific journal, The pathogenicity of the H5 subtype high pathogenicity avian influenza viruses (HPAIVs) in Ardeidae bird species has not been investigated yet, despite the increasing infections reported. Therefore, the present study aimed to examine the susceptibility of the Ardeidae species, which had already been reported to be susceptible to HPAIVs, to a clade 2.3.2.1 H5N1 HPAIV. Juvenile herons (four grey herons, one intermediate egret, two little egrets, and three black-crowned night herons) were intranasally inoculated with 106 50% egg infectious dose of the virus and observed for 10 days. Two of the four grey herons showed lethargy and conjunctivitis; among them, one died at 6 days post-inoculation (dpi). The viruses were transmitted to the other two cohoused naïve grey herons. Some little egrets and black-crowned night herons showing neurological disorders died at 4-5 dpi; these birds mainly shed the virus via the oral route. The viruses predominantly replicated in the brains of birds that died of infection. Seroconversion was observed in most surviving birds, except some black-crowned night herons. These results demonstrate that most Ardeidae species are susceptible to H5 HPAIVs, sometimes with lethal effects. Herons are mostly colonial and often share habitats with Anseriformes, natural hosts of influenza A viruses; therefore, the risks of cluster infection and contribution to viral dissemination should be continuously evaluated. RESEARCH HIGHLIGHTSClade 2.3.2.1 H5N1 HPAIV causes lethal infections in Ardeidae sp.Viruses are transmitted among grey herons.Some herons with HPAIV showed conjunctivitis or neurological symptoms.HPAIV systemically replicated in herons tissues.
• Antiviral Effects of 5-Aminolevulinic Acid Phosphate against Classical Swine Fever Virus: In Vitro and In Vivo Evaluation.
  Shizuka Hirose; Norikazu Isoda; Loc Tan Huynh; Taksoo Kim; Keiichiro Yoshimoto; Tohru Tanaka; Kenjiro Inui; Takahiro Hiono; Yoshihiro Sakoda
  Pathogens (Basel, Switzerland), 11, 2, 27 Jan. 2022, [International Magazine]
  English, Scientific journal, The inhibitory effects of 5-aminolevulinic acid phosphate (5-ALA), an important amino acid for energy production in the host, against viral infections were previously reported. Here, the antiviral effects of 5-ALA against classical swine fever virus (CSFV) belonging to the genus Pestivirus in the Flaviviridae family and its possible mechanisms were investigated. CSFV replication was suppressed in swine cells supplemented with 5-ALA or its metabolite, protoporphyrin IX (PPIX). The infectivity titer of CSFV was decreased after mixing with PPIX extracellularly. In addition, the activities of the replication cycle were decreased in the presence of PPIX based on the CSFV replicon assay. These results showed that PPIX exerted antiviral effects by inactivating virus particles and inhibiting the replication cycle. To evaluate the in vivo efficacy of 5-ALA, pigs were supplemented daily with 5-ALA for 1 week before virus inoculation and then inoculated with a virulent CSFV strain at the 107.0 50% tissue culture infectious dose. The clinical scores of the supplemented group were significantly lower than those of the nonsupplemented group, whereas the virus growth was not. Taken together, 5-ALA showed antiviral effects against CSFV in vitro, and PPIX played a key role by inactivating virus particles extracellularly and inhibiting the replication cycle intracellularly.
• Glycan Profiling of Viral Glycoproteins with the Lectin Microarray.
  Takahiro Hiono; Atsushi Kuno
  Methods in molecular biology (Clifton, N.J.), 2556, 59, 68, 2022, [Lead author, Corresponding author], [International Magazine]
  English, Scientific journal, Recently, structural analyses on the glycans attached to viral surface proteins have been intensively conducted since previous studies demonstrated that glycoform of the viral glycoproteins is closely related to their immunogenicity as vaccine antigens. Although mass spectrometric approach is a gold standard for the glycoproteomic analysis of viral glycoproteins, lectin microarray (LMA) is regarded as an alternative method for analyzing glycan attached to viruses. The previous studies demonstrated that LMA provides highly sensitive and straightforward platforms for the glycoproteomic analyses of viral glycans. Here, two methods, antibody-overlay method, and direct-labeling method, for profiling glycoforms of viral glycoprotein using LMA are described.
• Receptor-Binding Assay for Avian Influenza Viruses.
  Takahiro Hiono; Daiki Kobayashi
  Methods in molecular biology (Clifton, N.J.), 2556, 141, 148, 2022, [Lead author, Last author, Corresponding author], [International Magazine]
  English, Scientific journal, It is well known that influenza viruses utilize host cell glycans for virus attachment factors via their major glycoprotein, hemagglutinin (HA), to initiate their invasion to host cells. Unlike well-known theories in human and avian influenza viruses, barriers laying between interspecies transmission of influenza viruses among bird species are not well understood. Recently, it was speculated that glycan binding of the HA to fucosylated Siaα2-3Gal is related to the expansion in the host range of the virus in avian species. Accordingly, the binding specificity of avian influenza viruses to fucosylated Siaα2-3Gal glycans should be monitored for the better control of avian influenza in both poultry and wild birds. Here, general methods and points for the glycan-binding assay that are specifically modified to target fucosylated Siaα2-3Gal glycans are provided.
• Establishment of a mouse- and egg-adapted strain for the evaluation of vaccine potency against H3N2 variant influenza virus in mice.
  Enkhbold Bazarragchaa; Takahiro Hiono; Norikazu Isoda; Hirotaka Hayashi; Masatoshi Okamatsu; Yoshihiro Sakoda
  The Journal of veterinary medical science, 83, 11, 1694, 1701, 31 Oct. 2021, [Domestic magazines]
  English, Scientific journal, Sporadic spreads of swine-origin influenza H3N2 variant (H3N2v) viruses were reported in humans, resulting in 437 human infections between 2011 and 2021 in the USA. Thus, an effective vaccine is needed to better control a potential pandemic for these antigenically distinct viruses from seasonal influenza. In this study, a candidate vaccine strain with efficient growth capacity in chicken embryos was established through serial blind passaging of A/Indiana/08/2011 (H3N2)v in mice and chicken embryos. Seven amino acid substitutions (M21I in PA; A138T, N165K, and V226A in HA; S312L in NP; T167I in M1; G62A in NS1 proteins) were found in the passaged viruses without a major change in the antigenicity. This mouse- and egg-adapted virus was used as a vaccine and challenge strain in mice to evaluate the efficacy of the H3N2v vaccine in different doses. Antibodies with high neutralizing titers were induced in mice immunized with 100 µg of inactivated whole-virus particles, and those mice were significantly protected from the challenge of homologous strain. The findings indicated that the established strain in the study was useful for vaccine study in mouse models.
• Characteristics of Classical Swine Fever Virus Variants Derived from Live Attenuated GPE- Vaccine Seed.
  Taksoo Kim; Loc Tan Huynh; Shizuka Hirose; Manabu Igarashi; Takahiro Hiono; Norikazu Isoda; Yoshihiro Sakoda
  Viruses, 13, 8, 23 Aug. 2021, [International Magazine]
  English, Scientific journal, The GPE- strain is a live attenuated vaccine for classical swine fever (CSF) developed in Japan. In the context of increasing attention for the differentiating infected from vaccinated animals (DIVA) concept, the achievement of CSF eradication with the GPE- proposes it as a preferable backbone for a recombinant CSF marker vaccine. While its infectious cDNA clone, vGPE-, is well characterized, 10 amino acid substitutions were recognized in the genome, compared to the original GPE- vaccine seed. To clarify the GPE- seed availability, this study aimed to generate and characterize a clone possessing the identical amino acid sequence to the GPE- seed. The attempt resulted in the loss of the infectious GPE- seed clone production due to the impaired replication by an amino acid substitution in the viral polymerase NS5B. Accordingly, replication-competent GPE- seed variant clones were produced. Although they were mostly restricted to propagate in the tonsils of pigs, similarly to vGPE-, their type I interferon-inducing capacity was significantly lower than that of vGPE-. Taken together, vGPE- mainly retains ideal properties for the CSF vaccine, compared with the seed variants, and is probably useful in the development of a CSF marker vaccine.
• Application of Glycan-Related Microarrays
  Takahiro Hiono; Chiaki Nagai-Okatani; Atsushi Kuno
  Comprehensive Glycoscience: Second Edition, 134, 148, 21 Jun. 2021
  In book
• C-Terminally tagged NA in replication-competent influenza A viruses reveals differences in glycan profiles between NA and HA.
  Takahiro Hiono; Atsushi Kuno
  The Analyst, 145, 17, 5845, 5853, 24 Aug. 2020, [Lead author, Corresponding author], [International Magazine]
  English, Scientific journal, Glycans attached to the viruses regulate their pathogenicity, immunogenicity, and antigenicity. We have previously shown that lectin microarray provided an easy and highly sensitive platform for analyzing glycan profiles of hemagglutinin (HA) of influenza A viruses in culture supernatants. On the other hand, the system is not applicable for neuraminidase (NA), the other viral glycoprotein of influenza A viruses, due to the limited availability of specific antibodies used to detect NA in the lectin microarray. Accordingly, we established replication-competent viruses harboring the short peptide-tag sequence at the C-terminus of NA in this study. The generated viruses underwent normal proliferation cycles and showed similar properties to the wild-type viruses. Lectin microarray analyses of the tagged NA enriched from the viral particles showed that glycan profiles of NA were mostly occupied by mannose-type glycans. Interestingly, the profiles were distinct from those of HA separated from the same particle preparation, in which core-fucosylated complex-type N-glycans terminating with non-sialylated N-acetyllactosamine were dominant. Collectively, this study provides novel platforms for the analyses of the distinction between the glycan profiles of NA and HA, and contributes to a better understanding of later stages of the viral life cycles through analyzing the glycans attached to NA.
• Genetic and antigenic characterization of H5 and H7 avian influenza viruses isolated from migratory waterfowl in Mongolia from 2017 to 2019.
  Ankhanbaatar Ulaankhuu; Enkhbold Bazarragchaa; Masatoshi Okamatsu; Takahiro Hiono; Khishgee Bodisaikhan; Tsolmon Amartuvshin; Jargalsaikhan Tserenjav; Tsogtbaatar Urangoo; Khanui Buyantogtokh; Keita Matsuno; Takanari Hattori; Tatsunari Kondoh; Masahiro Sato; Yoshihiro Takadate; Shiho Torii; Mao Isono; Kosuke Okuya; Takeshi Saito; Nodoka Kasajima; Yurie Kida; Junki Maruyama; Manabu Igarashi; Ayato Takada; Hiroshi Kida; Damdinjav Batchuluun; Yoshihiro Sakoda
  Virus genes, 56, 4, 472, 479, Aug. 2020, [Peer-reviewed], [International Magazine]
  English, Scientific journal, The circulation of highly pathogenic avian influenza viruses (HPAIVs) of various subtypes (e.g., H5N1, H5N6, H5N8, and H7N9) in poultry remains a global concern for animal and public health. Migratory waterfowls play important roles in the transmission of these viruses across countries. To monitor virus spread by wild birds, active surveillance for avian influenza in migratory waterfowl was conducted in Mongolia from 2015 to 2019. In total, 5000 fecal samples were collected from lakesides in central Mongolia, and 167 influenza A viruses were isolated. Two H5N3, four H7N3, and two H7N7 viruses were characterized in this study. The amino acid sequence at hemagglutinin (HA) cleavage site of those isolates suggested low pathogenicity in chickens. Phylogenetic analysis revealed that all H5 and H7 viruses were closely related to recent H5 and H7 low pathogenic avian influenza viruses (LPAIVs) isolated from wild birds in Asia and Europe. Antigenicity of H7Nx was similar to those of typical non-pathogenic avian influenza viruses (AIVs). While HPAIVs or A/Anhui/1/2013 (H7N9)-related LPAIVs were not detected in migratory waterfowl in Mongolia, sporadic introductions of AIVs including H5 and H7 viruses into Mongolia through the wild bird migration were identified. Thus, continued monitoring of H5 and H7 AIVs in both domestic and wild birds is needed for the early detection of HPAIVs spread into the country.
• E190V substitution of H6 hemagglutinin is one of key factors for binding to sulfated sialylated glycan receptor and infection to chickens.
  Yuto Kikutani; Masatoshi Okamatsu; Shoko Nishihara; Sayaka Takase-Yoden; Takahiro Hiono; Robert P de Vries; Ryan McBride; Keita Matsuno; Hiroshi Kida; Yoshihiro Sakoda
  Microbiology and immunology, 64, 4, 304, 312, 14 Jan. 2020, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Avian influenza viruses (AIVs) recognize sialic acid linked α2,3 to galactose (SAα2,3Gal) glycans as receptors. In this study, the interactions between hemagglutinins (HAs) of AIVs and sulfated SAα2,3Gal glycans were analyzed to clarify the molecular basis of interspecies transmission of AIVs from ducks to chickens. It was revealed that E190V and N192D substitutions of the HA increased the recovery of viruses derived from an H6 duck virus isolate, A/duck/Hong Kong/960/1980 (H6N2), in chickens. Recombinant HAs from an H6 chicken virus, A/chicken/Tainan/V156/1999 (H6N1), bound to sulfated SAα2,3Gal glycans, whereas the HAs from an H6 duck virus did not. Binding preference of mutant HAs revealed that an E190V substitution is critical for the recognition of sulfated SAα2,3Gal glycans. These results suggest that the binding of the HA from H6 AIVs to sulfated SAα2,3Gal glycans explains a part of mechanisms of interspecies transmission of AIVs from ducks to chickens.
• Infection of newly identified phleboviruses in ticks and wild animals in Hokkaido, Japan indicating tick-borne life cycles.
  Shiho Torii; Keita Matsuno; Yongjin Qiu; Akina Mori-Kajihara; Masahiro Kajihara; Ryo Nakao; Naganori Nao; Katsunori Okazaki; Mariko Sashika; Takahiro Hiono; Masatoshi Okamatsu; Yoshihiro Sakoda; Hideki Ebihara; Ayato Takada; Hirofumi Sawa
  Ticks and tick-borne diseases, 10, 2, 328, 335, Feb. 2019, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Recent discoveries of tick-borne pathogens have raised public health concerns on tick-borne infectious diseases and emphasize the need to assess potential risks of unrecognized tick-borne pathogens. First, to determine the existence of tick-borne phleboviruses (TBPVs), genetic surveillance of phleboviruses in ticks was conducted mainly in Hokkaido, the northernmost island in Japan from 2013 to 2015. Genes of two TBPVs, previously reported as Mukawa virus (MKWV) and a newly identified relative of MKWV, Kuriyama virus (KURV), were detected and the viruses were isolated from Ixodes persulcatus collected in Hokkaido, but not in I. persulcatus collected from other areas of Japan. These viruses were phylogenetically and antigenically similar to each other. Next, to investigate the infection of MKWV in mammals, serum samples from wildlife captured in Hokkaido from 2007 to 2011 were used for serological screening. Neutralizing antibodies against MKWV were detected in both Yezo-deer (Cervus nippon yesoensis) (2/50) and raccoons (Procyon lotor) (16/64). However, no infectious MKWV was recovered from laboratory mice in experimental infections, though viral RNAs were detected in their tissues. Thus, MKWV and KURV may maintain tick-mammalian life cycles in Hokkaido, suggesting their potential as causative agents of tick-borne diseases in mammals.
• Lectin microarray analyses reveal host cell-specific glycan profiles of the hemagglutinins of influenza A viruses.
  Takahiro Hiono; Atsushi Matsuda; Takanori Wagatsuma; Masatoshi Okamatsu; Yoshihiro Sakoda; Atsushi Kuno
  Virology, 527, 132, 140, 15 Jan. 2019, [Peer-reviewed], [Lead author, Corresponding author], [International Magazine]
  English, Scientific journal, Glycan structures on hemagglutinin (HA) of influenza A viruses have been analyzed previously to understand their significance. However, the formerly established methods using mass spectrometry present disadvantages such as procedure complexity, sensitivity, and throughput. Our study has established a novel method for analyzing glycan profiles of HA using lectin microarray techniques. We successfully obtained glycan profiles of HA starting from 1 ml of the 106 TCID50 samples through simple antigen enrichment using optimized immunoprecipitation. The profiles were reasonably consistent with known glycan structures of HA. Next, we compared glycan profiles of the HAs prepared from chicken embryos, MDCK, Vero, and A549 cells, and demonstrated the host cell-specific HA glycan profiles. Notably, the HA from MDCK cells was α1-3 galactosylated. Our method provides a highly sensitive and simple procedure for glycan profiling of the viral glycoproteins, thereby paving way for direct glycan analyses of human- and animal-derived virions.
• H13 influenza viruses in wild birds have undergone genetic and antigenic diversification in nature.
  Zu-Jyun Wang; Yuto Kikutani; Lam Thanh Nguyen; Takahiro Hiono; Keita Matsuno; Masatoshi Okamatsu; Scott Krauss; Richard Webby; Youn-Jeong Lee; Hiroshi Kida; Yoshihiro Sakoda
  Virus genes, 54, 4, 543, 549, Aug. 2018, [Peer-reviewed], [International Magazine]
  English, Among 16 haemagglutinin (HA) subtypes of avian influenza viruses (AIVs), H13 AIVs have rarely been isolated in wild waterfowl. H13 AIVs cause asymptomatic infection and are maintained mainly in gull and tern populations; however, the recorded antigenic information relating to the viruses has been limited. In this study, 2 H13 AIVs, A/duck/Hokkaido/W345/2012 (H13N2) and A/duck/Hokkaido/WZ68/2012 (H13N2), isolated from the same area in the same year in our surveillance, were genetically and antigenically analyzed with 10 representative H13 strains including a prototype strain, A/gull/Maryland/704/1977 (H13N6). The HA genes of H13 AIVs were phylogenetically divided into 3 groups (I, II, and III). A/duck/Hokkaido/W345/2012 (H13N2) was genetically classified into Group III. This virus was distinct from a prototype strain, A/gull/Maryland/704/1977 (H13N6), and the virus, A/duck/Hokkaido/WZ68/2012 (H13N2), both belonging to Group I. Antigenic analysis indicated that the viruses of Group I were antigenically closely related to those of Group II, but distinct from those of Group III, including A/duck/Hokkaido/W345/2012 (H13N2). In summary, our study indicates that H13 AIVs have undergone antigenic diversification in nature.
• Isolation and characterization of avian influenza viruses from raw poultry products illegally imported to Japan by international flight passengers
  A. Shibata; T. Hiono; H. Fukuhara; R. Sumiyoshi; A. Ohkawara; K. Matsuno; M. Okamatsu; H. Osaka; Y. Sakoda
  Transboundary and Emerging Diseases, 65, 2, 465, 475, Blackwell Publishing Ltd, 01 Apr. 2018, [Peer-reviewed], [Lead author]
  English, Scientific journal
• Potency of an inactivated influenza vaccine prepared from A/duck/Mongolia/245/2015 (H10N3) against H10 influenza virus infection in a mouse model
  Mizuho Suzuki; Masatoshi Okamatsu; Yuri Fujimoto; Takahiro Hiono; Keita Matsuno; Hiroshi Kida; Yoshihiro Sakoda
  Japanese Journal of Veterinary Research, 66, 1, 29, 41, Hokkaido University, 2018, [Peer-reviewed]
  English, Scientific journal
• Potency of an inactivated influenza vaccine prepared from A/duck/Hokkaido/162/2013 (H2N1) against a challenge with A/swine/Missouri/2124514/2006 (H2N3) in mice
  Mizuho Suzuki; Masatoshi Okamatsu; Takahiro Hiono; Keita Matsuno; Yoshihiro Sakoda
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 79, 11, 1815, 1821, Nov. 2017, [Peer-reviewed]
  English, Scientific journal
• Selection of antigenic variants of an H5N1 highly pathogenic avian influenza virus in vaccinated chickens
  Lam Thanh Nguyen; Tatsuya Nishi; Shintaro Shichinohe; Duc-Huy Chu; Takahiro Hiono; Keita Matsuno; Masatoshi Okamatsu; Hiroshi Kida; Yoshihiro Sakoda
  VIROLOGY, 510, 252, 261, Oct. 2017, [Peer-reviewed]
  English, Scientific journal
• Characterization of H5N6 highly pathogenic avian influenza viruses isolated from wild and captive birds in the winter season of 2016-2017 in Northern Japan
  Takahiro Hiono; Masatoshi Okamatsu; Keita Matsuno; Atsushi Haga; Ritsuko Iwata; Lam Thanh Nguyen; Mizuho Suzuki; Yuto Kikutani; Hiroshi Kida; Manabu Onuma; Yoshihiro Sakoda
  MICROBIOLOGY AND IMMUNOLOGY, 61, 9, 387, 397, Sep. 2017, [Peer-reviewed], [Lead author]
  English, Scientific journal
• Rapid and broad detection of H5 hemagglutinin by an immunochromatographic kit using novel monoclonal antibody against highly pathogenic avian influenza virus belonging to the genetic clade 2.3.4.4
  Lam Thanh Nguyen; Kazunari Nakaishi; Keiko Motojima; Ayako Ohkawara; Erina Minato; Junki Maruyama; Takahiro Hiono; Keita Matsuno; Masatoshi Okamatsu; Takashi Kimura; Ayato Takada; Hiroshi Kida; Yoshihiro Sakoda
  PLOS ONE, 12, 8, e0182228, Aug. 2017, [Peer-reviewed]
  English, Scientific journal
• Genetic and virulence characterization of classical swine fever viruses isolated in Mongolia from 2007 to 2015
  Bazarragchaa Enkhbold; Munkhduuren Shatar; Shiho Wakamori; Tomokazu Tamura; Takahiro Hiono; Keita Matsuno; Masatoshi Okamatsu; Takashi Umemura; Batchuluun Damdinjav; Yoshihiro Sakoda
  VIRUS GENES, 53, 3, 418, 425, Jun. 2017, [Peer-reviewed]
  English, Scientific journal
• Antigenic diversity of H5 highly pathogenic avian influenza viruses of clade 2.3.4.4 isolated in Asia
  Ayako Ohkawara; Masatoshi Okamatsu; Makoto Ozawa; Duc-Huy Chu; Lam Thanh Nguyen; Takahiro Hiono; Keita Matsuno; Hiroshi Kida; Yoshihiro Sakoda
  MICROBIOLOGY AND IMMUNOLOGY, 61, 5, 149, 158, May 2017, [Peer-reviewed]
  English, Scientific journal
• Potency of whole virus particle and split virion vaccines using dissolving microneedle against challenges of H1N1 and H5N1 influenza viruses in mice
  Akihiro Nakatsukasa; Koji Kuruma; Masatoshi Okamatsu; Takahiro Hiono; Mizuho Suzuki; Keita Matsuno; Hiroshi Kida; Takayoshi Oyamada; Yoshihiro Sakoda
  VACCINE, 35, 21, 2855, 2861, May 2017, [Peer-reviewed]
  English, Scientific journal
• Characterization of Highly Pathogenic Avian Influenza Virus A(H5N6), Japan, November 2016
  Masatoshi Okamatsu; Makoto Ozawa; Kosuke Soda; Hiroki Takakuwa; Atsushi Haga; Takahiro Hiono; Aya Matsuu; Yuko Uchida; Ritsuko Iwata; Keita Matsuno; Masakazu Kuwahara; Toshiyo Yabuta; Tatsufumi Usui; Hiroshi Ito; Manabu Onuma; Yoshihiro Sakoda; Takehiko Saito; Koichi Otsuki; Toshihiro Ito; Hiroshi Kida
  EMERGING INFECTIOUS DISEASES, 23, 4, 691, 695, Apr. 2017, [Peer-reviewed]
  English, Scientific journal
• Nationwide Distribution of Bovine Influenza D Virus Infection in Japan
  Taisuke Horimoto; Takahiro Hiono; Hirohisa Mekata; Tomoha Odagiri; Zhihao Lei; Tomoya Kobayashi; Junzo Norimine; Yasuo Inoshima; Hirokazu Hikono; Kenji Murakami; Reiichiro Sato; Hironobu Murakami; Masahiro Sakaguchi; Kazunori Ishii; Takaaki Ando; Kounosuke Otomaru; Makoto Ozawa; Yoshihiro Sakoda; Shin Murakami
  PLOS ONE, 11, 9, e0163828, Sep. 2016, [Peer-reviewed]
  English, Scientific journal
• Recent developments in the diagnosis of avian influenza
  Masatoshi Okamatsu; Takahiro Hiono; Hiroshi Kida; Yoshihiro Sakoda
  VETERINARY JOURNAL, 215, 82, 86, Sep. 2016, [Peer-reviewed]
  English
• Genetic and antigenic characterization of H5, H6 and H9 avian influenza viruses circulating in live bird markets with intervention in the center part of Vietnam
  Duc-Huy Chu; Masatoshi Okamatsu; Keita Matsuno; Takahiro Hiono; Kohei Ogasawara; Lam Thanh Nguyen; Long Van Nguyen; Tien Ngoc Nguyen; Thuy Thu Nguyen; Dong Van Pham; Dang Hoang Nguyen; Tho Dang Nguyen; Thanh Long To; Hung Van Nguyen; Hiroshi Kida; Yoshihiro Sakoda
  VETERINARY MICROBIOLOGY, 192, 194, 203, Aug. 2016, [Peer-reviewed]
  English, Scientific journal
• Is the optimal pH for membrane fusion in host cells by avian influenza viruses related to host range and pathogenicity?
  Masatoshi Okamatsu; Yurie Motohashi; Takahiro Hiono; Tomokazu Tamura; Kazuki Nagaya; Keita Matsuno; Yoshihiro Sakoda; Hiroshi Kida
  ARCHIVES OF VIROLOGY, 161, 8, 2235, 2242, Aug. 2016, [Peer-reviewed]
  English, Scientific journal
• Cytokine responses to eye spray adjuvants for enhancing vaccine-induced immunity in chickens.
  Hiromi Takaki; Haruko Sato; Riho Kurata; Hirokazu Hikono; Takahiro Hiono; Hiroshi Kida; Misako Matsumoto; Takehiko Saito; Tsukasa Seya
  Microbiology and immunology, 60, 7, 511, 5, Jul. 2016, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Eye spray influenza vaccines for chickens are increasingly available; however, how to enhance cellular and antibody responses to them remains undetermined. Here, eye-drops containing the immune-enhancing adjuvants Pam2CSK4 or polyI:C were assessed in chickens. Application of these TLR agonists to chicken conjunctiva resulted in up-regulation of IL-1β, but not other cytokines, including IFN and IL-6, in the spleen, lung and Harderian gland. Thus, responses to adjuvant applied to the conjunctival mucosa of chickens differ from those expected from the responses to intra-nasal adjuvants in mammals. Identifying an appropriate delivery route for adjuvants is crucial for evoking immune responses in chickens.
• Amino acid residues at positions 222 and 227 of the hemagglutinin together with the neuraminidase determine binding of H5 avian influenza viruses to sialyl Lewis X
  Takahiro Hiono; Masatoshi Okamatsu; Manabu Igarashi; Ryan McBride; Robert P. de Vries; Wenjie Peng; James C. Paulson; Yoshihiro Sakoda; Hiroshi Kida
  ARCHIVES OF VIROLOGY, 161, 2, 307, 316, Feb. 2016, [Peer-reviewed], [Lead author]
  English, Scientific journal
• Complete genome sequence of the avian paramyxovirus serotype 5 strain APMV-5/budgerigar/Japan/TI/75
  Takahiro Hiono; Keita Matsuno; Kotaro Tuchiya; Zhifeng Lin; Masatoshi Okamatsu; Yoshihiro Sakoda
  Genome Announcements, 4, 5, American Society for Microbiology, 2016, [Peer-reviewed], [Lead author]
  English, Scientific journal
• Experimental infection of highly and low pathogenic avian influenza viruses to chicken's, ducks, tree sparrows, jungle crows, and black rats for the evaluation of their roles in virus transmission
  Takahiro Hiono; Masatoshi Okamatsu; Naoki Yamamoto; Kohei Ogasawara; Mayumi Endo; Saya Kuribayashi; Shintaro Shichinohe; Yurie Motohashi; Duc-Huy Chu; Mizuho Suzuki; Takaya Ichikawa; Tatsuya Nishi; Yuri Abe; Keita Matsuno; Kazuyuki Tanaka; Tsutomu Tanigawa; Hiroshi Kida; Yoshihiro Sakoda
  VETERINARY MICROBIOLOGY, 182, 108, 115, Jan. 2016, [Peer-reviewed], [Lead author]
  English, Scientific journal
• Genetic and antigenic characterization of H5 and H7 influenza viruses isolated from migratory water birds in Hokkaido, Japan and Mongolia from 2010 to 2014
  Takahiro Hiono; Ayako Ohkawara; Kohei Ogasawara; Masatoshi Okamatsu; Tomokazu Tamura; Duc-Huy Chu; Mizuho Suzuki; Saya Kuribayashi; Shintaro Shichinohe; Ayato Takada; Hirohito Ogawa; Reiko Yoshida; Hiroko Miyamoto; Naganori Nao; Wakako Furuyama; Junki Maruyama; Nao Eguchi; Gerelmaa Ulziibat; Bazarragchaa Enkhbold; Munkhduuren Shatar; Tserenjav Jargalsaikhan; Selenge Byambadorj; Batchuluun Damdinjav; Yoshihiro Sakoda; Hiroshi Kida
  VIRUS GENES, 51, 1, 57, 68, Aug. 2015, [Peer-reviewed], [Lead author]
  English, Scientific journal
• Potency of an inactivated influenza vaccine prepared from A/duck/Mongolia/119/2008 (H7N9) against the challenge with A/Anhui/1/2013 (H7N9)
  Duc-Huy Chu; Yoshihiro Sakoda; Tatsuya Nishi; Takahiro Hiono; Shintaro Shichinohe; Masatoshi Okamatsu; Hiroshi Kida
  VACCINE, 32, 28, 3473, 3479, Jun. 2014, [Peer-reviewed]
  English, Scientific journal
• Protective Efficacy of Passive Immunization with Monoclonal Antibodies in Animal Models of H5N1 Highly Pathogenic Avian Influenza Virus Infection
  Yasushi Itoh; Reiko Yoshida; Shintaro Shichinohe; Megumi Higuchi; Hirohito Ishigaki; Misako Nakayama; Van Loi Pham; Hideaki Ishida; Mitsutaka Kitano; Masahiko Arikata; Naoko Kitagawa; Yachiyo Mitsuishi; Kazumasa Ogasawara; Hideaki Tsuchiya; Takahiro Hiono; Masatoshi Okamatsu; Yoshihiro Sakoda; Hiroshi Kida; Mutsumi Ito; Le Quynh Mai; Yoshihiro Kawaoka; Hiroko Miyamoto; Mari Ishijima; Manabu Igarashi; Yasuhiko Suzuki; Ayato Takada
  PLOS PATHOGENS, 10, 6, e1004192, Jun. 2014, [Peer-reviewed]
  English, Scientific journal
• A chicken influenza virus recognizes fucosylated alpha 2,3 sialoglycan receptors on the epithelial cells lining upper respiratory tracts of chickens
  Takahiro Hiono; Masatoshi Okamatsu; Shoko Nishihara; Sayaka Takase-Yoden; Yoshihiro Sakoda; Hiroshi Kida
  VIROLOGY, 456, 131, 138, May 2014, [Peer-reviewed], [Lead author]
  English, Scientific journal
• Potency of a vaccine prepared from A/swine/Hokkaido/2/1981 (H1N1) against A/Narita/1/2009 (H1N1) pandemic influenza virus strain
  Masatoshi Okamatsu; Yoshihiro Sakoda; Takahiro Hiono; Naoki Yamamoto; Hiroshi Kida
  VIROLOGY JOURNAL, 10, 47, Feb. 2013, [Peer-reviewed]
  English, Scientific journal

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• ワクチン非接種牛群における牛ウイルス性下痢ウイルスに対する抗体陽性率の算出および感染力の推定
  磯田典和; 磯田典和; 磯田典和; 磯田典和; 関口敏; 関口敏; HUNG Ngo Dinh; HUNG Ngo Dinh; 龍千香; 牛谷雄一; 野津昂亮; 濱口華凜; 小林茉弥; 日尾野隆大; 迫田義博; 日尾野隆大; 迫田義博; 日尾野隆大; 迫田義博; 日尾野隆大; 迫田義博, 日本獣医学会学術集会講演要旨集, 167th, 2024
• ノイラミニダーゼストーク領域におけるN型糖鎖欠損がH7N7高病原性鳥インフルエンザウイルスのニワトリに対する病原性を高める
  小林大樹; 日尾野隆大; 日尾野隆大; 日尾野隆大; 日尾野隆大; 荒川広夢; 梶裕之; 大河原彩子; 市川貴也; 伴日向子; 磯田典和; 迫田義博; 磯田典和; 迫田義博; 磯田典和; 迫田義博; 磯田典和; 迫田義博, 日本獣医学会学術集会講演要旨集, 167th, 2024
• Pathogenicity of different genotypes of Clade 2.3.4.4b H5N1 high pathogenicity avian influenza virus in a mammalian model
  七戸新太郎; 日尾野隆大; 日尾野隆大; 日尾野隆大; 下岡誠; 磯田典和; 磯田典和; 磯田典和; 迫田義博; 迫田義博; 迫田義博; 迫田義博; 渡辺登喜子; 渡辺登喜子; 渡辺登喜子, 日本ウイルス学会学術集会プログラム・予稿集(Web), 71st, 2024
• Establishment of a drug deployment system for the treatment of high pathogenicity avian influenza virus infections in rare birds: efforts towards biodiversity conservation and promoting proper use of medications
  迫田義博; 迫田義博; 日尾野隆大; 日尾野隆大; 池中良徳; 池中良徳, 日本野生動物医学会大会・講演要旨集, 30th, 2024
• Pharmacokinetic analysis of baloxavir marboxil in captive rare birds to establish the treatment protocol against high pathogenicity avian influenza virus infection
  島津陽; 三木万梨子; 尾原涼; 中谷裕美子; 長嶺隆; 吉本悠人; 林万里菜; 井出いづみ; 見浦沙耶子; 平栗祐子; 加来雅人; 豊田恒介; 尾崎美樹; 日尾野隆大; 日尾野隆大; 磯田典和; 磯田典和; 宍戸貴雄; 池中良徳; 迫田義博; 迫田義博, 日本野生動物医学会大会・講演要旨集, 30th, 2024
• Study of the administration route of baloxavir marboxil in non-clinical models-Aiming to improve absorption in the Okinawa rail-
  三木万梨子; 島津陽; 尾原涼; 大崎裕美; 宍戸貴雄; 池中良徳; 木村享史; 日尾野隆大; 日尾野隆大; 磯田典和; 磯田典和; 迫田義博; 迫田義博, 日本野生動物医学会大会・講演要旨集, 30th, 2024
• インフルエンザAウイルスのヘマグルチニンに対する高親和性及び低親和性細胞の作製
  前川明博; 日尾野隆大; 迫田義博; 喜田宏; 栂谷内晶; 栂谷内晶; 西原祥子; 西原祥子; 高瀬明; 高瀬明, 日本分子生物学会年会プログラム・要旨集(Web), 47th, 2024
• Elucidation of the glycan structure that contributes to bind of influenza A virus using glycan engineered cells.
  前川明博; 日尾野隆大; 迫田義博; 喜田宏; 栂谷内晶; 栂谷内晶; 西原祥子; 西原祥子; 高瀬明; 高瀬明, 日本ウイルス学会学術集会プログラム・予稿集(Web), 71st, 2024
• [The current situation of H5 high pathogenicity avian influenza viruses in wild birds and mammals].
  Takahiro Hiono; Norikazu Isoda; Yoshihiro Sakoda, Uirusu, 74, 2, 107, 116, 2024, [Domestic magazines]
  H5 high pathogenicity avian influenza viruses, which emerged in Guangdong Province, China, in 1996, has now been persistently transmitted among various wild birds due to the "silent spreading" of the viruses among vaccinated poultry and domestic waterfowl. These viruses traveled long distances along with bird migration; therefore, the threat of H5 high pathogenicity avian influenza viruses is now a global issue. Furthermore, infection in wild mammals has become more prominent since 2020. The contamination of the wild bird population by the virus is considered to be an irreversible situation, and thus, the reduction of virus levels in the environment is an urgent issue to prevent further deterioration of the situation. This review will describe the history and current situations of influenza virus infection in wild birds and mammals, and discuss the research and countermeasures that are required to stop the damage caused by this virus., Japanese
• 2021-2022年冬季に国内で検出されたH5亜型高病原性鳥インフルエンザウイルスの性状と来シーズンの展望
  迫田 義博; 日尾野 隆大; 小林 大樹; Gulyaeva Marina; Shestopalov Alexander; 鍋島 圭; 本庄 比佐子; 横山 美沙子; 大沼 学; 磯田 典和, 日本野生動物医学会誌, 27, Suppl., 161, 161, Mar. 2023
  日本野生動物医学会, Japanese
• 鳥類におけるバロキサビルマルボキシルの安全性の検討
  三木 万梨子; 尾原 涼; 西村 享平; 岡 良子; 宍戸 貴雄; 福島 民雄; 吉本 淳; 中山 翔太; 木村 亨史; 池中 良徳; 小笠原 浩平; 齊藤 慶輔; 日尾野 隆大; 磯田 典和; 迫田 義博, 日本野生動物医学会誌, 27, Suppl., 176, 177, Mar. 2023
  日本野生動物医学会, Japanese
• 高病原性鳥インフルエンザウイルスに感染したオジロワシにおける,バロキサビルマルボキシルによる治療の試み
  齊藤 慶輔; 渡邊 有希子; 小笠原 浩平; 磯田 典和; 日尾野 隆大; 宍戸 貴雄; 西村 享平; 安達 光; 河野 晴子; 石井 千尋; 大沼 学; 迫田 義博, 日本野生動物医学会誌, 27, Suppl., 182, 182, Mar. 2023
  日本野生動物医学会, Japanese
• 高病原性鳥インフルエンザウイルスの現状とOne Health
  日尾野隆大; 日尾野隆大; 日尾野隆大; 磯田典和; 磯田典和; 磯田典和; 磯田典和; 迫田義博; 迫田義博; 迫田義博; 迫田義博, 人と動物の共通感染症研究会学術集会講演要旨集(Web), 23rd, 2023
• 北海道における野鳥および野生哺乳動物からのH5N1亜型高病原性鳥インフルエンザウイルスの検出事例について
  磯田 典和; 日尾野 隆大; 迫田 義博, 病原微生物検出情報月報, 43, 11, 259, 260, Nov. 2022
  国立感染症研究所, Japanese
• 2021～2022年シーズンの野鳥における高病原性鳥インフルエンザウイルスの流行の特徴と考察
  小笠原 浩平; 渡辺 有希子; 磯田 典和; 日尾野 隆大; 安達 光; 河野 晴子; 大沼 学; 迫田 義博; 齊藤 慶輔, 北海道獣医師会雑誌, 66, 8, 302, 302, Aug. 2022
  (公社)北海道獣医師会, Japanese
• 2014年から2020年に日本で分離された牛ウイルス性下痢ウイルスの遺伝子解析
  荻野紗帆; 西森朝美; 廣瀬静香; 磯田典和; 磯田典和; 日尾野隆大; 安藤清彦; 迫田義博; 迫田義博, 日本獣医学会学術集会講演要旨集, 164th (CD-ROM), 2021
• シチメンチョウの気管上皮に発現する多様な鳥インフルエンザウイルスレセプターの構造解析
  小林大樹; 市居修; 日尾野隆大; 西原祥子; 高瀬明; 磯田典和; 磯田典和; 迫田義博; 迫田義博, 日本獣医学会学術集会講演要旨集, 164th (CD-ROM), 2021
• 2020年度冬季に東日本の野鳥から分離された高病原性鳥インフルエンザウイルスの性状
  磯田典和; 磯田典和; 日尾野隆大; TWABELA Augustin T.; BAZARRAGCHAA Enkhbold; KIEN Le Trung; LOC Huynh Tan; 小笠原浩平; 林裕貴; 渡辺有希子; 齊藤慶輔; 喜田宏; 迫田義博; 迫田義博, 日本獣医学会学術集会講演要旨集, 164th (CD-ROM), 2021
• アミノレブリン酸リン酸塩による豚熱ウイルス増殖抑制効果の検証
  廣瀬静香; LOC Huynh Tan; 金琢珠; 磯田典和; 磯田典和; 日尾野隆大; 吉本圭一郎; 田中徹; 乾健二郎; 迫田義博; 迫田義博, 日本獣医学会学術集会講演要旨集, 164th (CD-ROM), 2021
• ワクチンシードと同じアミノ酸配列を持つ豚熱ウイルスGPE^-株作出の試み
  金琢洙; LOC Huynh Tan; 廣瀬静香; 日尾野隆大; 磯田典和; 磯田典和; 迫田義博; 迫田義博, 日本獣医学会学術集会講演要旨集, 164th (CD-ROM), 2021
• Generation of recombinant highly virulent classical swine fever virus strain possessing a luminescent HiBiT tag as a reporter
  LOC Huynh Tan; 廣瀬静香; 金琢洙; 田村友和; 福原崇介; 松浦善治; 日尾野隆大; 磯田典和; 磯田典和; 迫田義博; 迫田義博, 日本獣医学会学術集会講演要旨集, 164th (CD-ROM), 2021
• The pioneering study on impact identification of low pathogenicity avian influenza virus infection in the farm in southern Vietnam
  LE Trung Kien; 磯田典和; 磯田典和; NGUYEN Thanh Lam; NGUYEN Thanh Lam; CHU Duc-Huy; TIEN Ngoc Tien; LE Thanh Tung; 日尾野隆大; 迫田義博; 迫田義博, 日本獣医学会学術集会講演要旨集, 164th (CD-ROM), 2021
• 豚熱ウイルスに対する各種動物用消毒薬の効果
  迫田義博; 迫田義博; 遠藤真由美; 伊藤貢; 日尾野隆大; 磯田典和; 磯田典和, 日本獣医学会学術集会講演要旨集, 164th (CD-ROM), 2021
• 豚熱ウイルスに対する各種動物用消毒薬の効果
  迫田義博; 迫田義博; 遠藤真由美; 伊藤貢; 日尾野隆大; 磯田典和; 磯田典和, 家畜衛生学雑誌, 47, 3, 2021
• 質量分析とレクチンアレイを組み合わせたSARS-CoV-2スパイクタンパク質の高精度糖鎖構造解析
  日尾野隆大; 日尾野隆大; 富岡あづさ; 梶裕之; 佐々木道仁; 大場靖子; 澤洋文; 久野敦, 日本獣医学会学術集会講演要旨集, 164th (CD-ROM), 2021
• 鳥インフルエンザウイルスの中間宿主としてのウズラおよびシチメンチョウにおけるシアル酸糖鎖分布の解析
  小林大樹; 佐々木乙; 日尾野隆大; 岡松正敏; 西原祥子; 高瀬明; 市居修; 松野啓太; 松野啓太; 迫田義博; 迫田義博, 日本獣医学会学術集会講演要旨集, 163rd, 2020
• Does the receptor-binding specificity of influenza viruses explain their tissue tropism in hosts?
  Takahiro Hiono; Masatoshi Okamatsu; Yoshihiro Sakoda, Clinical Virology, 47, 1, 61, 68, Mar. 2019, [Peer-reviewed], [Invited]
  日本臨床ウイルス学会, Japanese, Introduction scientific journal
• Influenza viruses in glycoscience
  Takahiro Hiono, BIO Clinica, 34, 2, 79, 85, Feb. 2019, [Peer-reviewed], [Invited]
  Japanese, Introduction commerce magazine
• 2016‐2017年冬季に北日本で野鳥及び飼育鳥から分離されたH5N6高病原性鳥インフルエンザウイルスの性状解析
  岡松正敏; 日尾野隆大; 菊谷祐斗; 鈴木瑞穂; NGUYEN Thanh‐Lam; 松野啓太; 松野啓太; 迫田義博; 迫田義博, 日本獣医学会学術集会講演要旨集, 160th, 382, 382, 30 Aug. 2017
  (公社)日本獣医学会, Japanese
• 2016‐2017年冬季に北日本で野鳥及び飼育鳥から分離されたH5N6高病原性鳥インフルエンザウイルスの性状解析
  岡松正敏; 日尾野隆大; 菊谷祐斗; 鈴木瑞穂; NGUYEN Thanh‐Lam; 松野啓太; 松野啓太; 迫田義博; 迫田義博, 日本獣医学会学術集会講演要旨集, 160th, 382, 382, 30 Aug. 2017
  (公社)日本獣医学会, Japanese
• 2.3.4.4高病原性鳥インフルエンザウイルスに対する新規モノクローナル抗体を用いた免疫クロマトグラフィーキットによるH5ヘマグルチニンの迅速かつ広範囲検出(Rapid and broad detection of H5 hemagglutinin by an immunochromatographic kit using novel monoclonal antibody against a 2.3.4.4 highly pathogenic avian influenza virus)
  Nguyen Thanh-Lam; 中石 和成; 本島 桂子; 大河原 彩子; 日尾野 隆大; 松野 啓太; 岡松 正敏; 高田 礼人; 喜田 宏; 迫田 義博, 日本獣医学会学術集会講演要旨集, 160回, 384, 384, Aug. 2017
  (公社)日本獣医学会, English
• Rapid and broad detection of H5 hemagglutinin by an immunochromatographic kit using novel monoclonal antibody against a 2.3.4.4 highly pathogenic avian influenza virus(和訳中)
  Nguyen Thanh-Lam; 中石 和成; 本島 桂子; 大河原 彩子; 日尾野 隆大; 松野 啓太; 岡松 正敏; 高田 礼人; 喜田 宏; 迫田 義博, 日本獣医学会学術集会講演要旨集, 160回, 384, 384, Aug. 2017
  (公社)日本獣医学会, English
• モンゴルにおける豚コレラの発生と分離ウイルスの遺伝子と病原性の解析
  迫田義博; 迫田義博; BAZARRAGCHAA Enkhbold; MUNKHDUUREN Shatar; 若森史穂; 日尾野隆大; 松野啓太; 松野啓太; 岡松正敏; BATCHULUUN Damdinjav; 梅村孝司, 日本獣医学会学術集会講演要旨集, 159th, 388, 388, 30 Aug. 2016
  (公社)日本獣医学会, Japanese
• D型インフルエンザウイルスはわが国のウシ社会に侵淫している
  堀本泰介; 日尾野隆大; 目堅博久; 小田切友葉; 雷志皓; 遠藤麻衣子; 上間亜希子; 小林知也; CHAMBERS James; 内田和幸; 西原真杉; 乗峰潤三; 猪島康雄; 彦野弘一; 村上賢二; 佐藤礼一郎; 村上裕信; 阪口雅弘; 安藤貴朗; 乙丸孝之介; 小澤真; 石井一功; 迫田義博; 村上晋, 日本獣医学会学術集会講演要旨集, 159th, 386, 386, 30 Aug. 2016
  (公社)日本獣医学会, Japanese
• 海外から持ち込まれた未加熱家きん肉から分離された鳥インフルエンザウイルスの性状解析
  柴田明弘; 日尾野隆大; 福原久江; 住吉理穂; 大河原彩子; 松野啓太; 松野啓太; 岡松正敏; 迫田義博; 迫田義博; 尾坂優之, 日本獣医学会学術集会講演要旨集, 159th, 382, 382, 30 Aug. 2016
  (公社)日本獣医学会, Japanese
• ワクチン接種後の鶏におけるH5鳥インフルエンザウイルスの抗原多様性の選択(Selection of antigenic variants of H5 avian influenza viruses in vaccinated chickens)
  Nguyen Thanh-Lam; 西 達也; 七戸 新太郎; Chu Duc-Huy; 日尾野 隆大; 松野 啓太; 岡松 正敏; 喜田 宏; 迫田 義博, 日本獣医学会学術集会講演要旨集, 159回, 371, 371, Aug. 2016
  (公社)日本獣医学会, English
• インフルエンザウイルスのフコシル化α2,3シアロ糖認識機構の解析
  日尾野隆大; 岡松正敏; 五十嵐学; 五十嵐学; MCBRIDE Ryan; DE VIRES Robert; PAULSON James; 松野啓太; 松野啓太; 迫田義博; 迫田義博; 喜田宏; 喜田宏; 喜田宏, 日本獣医学会学術集会講演要旨集, 158th, 332, 332, 30 Aug. 2015
  (公社)日本獣医学会, Japanese
• 新規開発マイクロニードルを用いた貼るインフルエンザワクチンの効果
  中務陽裕; 来馬浩二; 鈴木瑞穂; 日尾野隆大; 小山田孝嘉; 岡松正敏; 迫田義博; 迫田義博, 日本獣医学会学術集会講演要旨集, 158th, 329, 329, 30 Aug. 2015
  (公社)日本獣医学会, Japanese
• 近年アジア地域で分離された鳥インフルエンザウイルスのアヒル,スズメ,カラス,およびクマネズミに対する病原性
  小笠原浩平; 岡松正敏; 日尾野隆大; 七戸新太郎; 栗林沙弥; 市川貴也; 西達也; CHU Duc‐Huy; 鈴木瑞穂; 遠藤真由美; 田中和之; 谷川力; 喜田宏; 喜田宏; 喜田宏; 迫田義博; 迫田義博; 迫田義博, 日本獣医学会学術集会講演要旨集, 158th, 333, 333, 30 Aug. 2015
  (公社)日本獣医学会, Japanese
• 2014年のベトナムにおける介入有りまたは介入なしの生鳥市場にて単離された鳥インフルエンザウイルスの性状解析(Characterization of avian influenza viruses isolated in live bird markets with and without intervention in Vietnam in 2014)
  Chu Duc-Huy; 岡松 正敏; 松野 啓太; 日尾野 隆大; 小笠原 浩平; 鈴木 瑞穂; Nguyen Thanh Lam; 喜田 宏; 迫田 義博, 日本獣医学会学術集会講演要旨集, 158回, 329, 329, Aug. 2015
  (公社)日本獣医学会, English
• インフルエンザウイルスのフコシル化α2,3シアロ糖認識機構の解析
  日尾野隆大; 岡松正敏; 五十嵐学; 五十嵐学; MCBRIDE Ryan; DE VIRES Robert P; PAULSON James C; 迫田義博; 迫田義博; 喜田宏; 喜田宏, 日本糖質学会年会要旨集, 34th, 173, 01 Jul. 2015
  Japanese
• 鳥インフルエンザウイルスのヒト型レセプター特異性獲得メカニズムの解析
  柳田里澄; 岡松正敏; 日尾野隆大; 迫田義博; 喜田宏, 日本ウイルス学会学術集会プログラム・抄録集, 62nd, 155, 31 Oct. 2014
  Japanese
• ニワトリのインフルエンザウイルスはフコシル化および硫酸化α2,3シアル酸糖鎖をレセプターとする
  岡松正敏; 日尾野隆大; 五十嵐学; DE VRIES Robert; 西原祥子; 高瀬明; PAULSON James C; 迫田義博; 喜田宏, 日本ウイルス学会学術集会プログラム・抄録集, 62nd, 154, 31 Oct. 2014
  Japanese
• H5N1インフルエンザウイルスに対する中和抗体を用いた受動免疫の感染防御効果
  吉田玲子; 伊藤靖; 七戸新太郎; 小笠原一誠; 日尾野隆大; 岡松正敏; 迫田義博; 喜田宏; LE Mai Quynh; 河岡義裕; 五十嵐学; 鈴木定彦; 高田礼人, 日本ウイルス学会学術集会プログラム・抄録集, 62nd, 234, 31 Oct. 2014
  Japanese
• H5N1高病原性鳥インフルエンザウイルスがブタに感染するとSAα2,6Galレセプターに結合するウイルスが選択されるか?
  七戸新太郎; 迫田義博; 西達也; 日尾野隆大; 岡松正敏; 伊藤靖; 小笠原一誠; 喜田宏, 日本ウイルス学会学術集会プログラム・抄録集, 62nd, 154, 31 Oct. 2014
  Japanese
• 2010年から2014年に家禽及び野鳥から分離されたH5及びH7鳥インフルエンザウイルスの遺伝子と抗原性の解析
  迫田義博; 大河原彩子; 日尾野隆大; 小笠原浩平; 岡松正敏; 喜田宏, 日本ウイルス学会学術集会プログラム・抄録集, 62nd, 163, 31 Oct. 2014
  Japanese
• 2010年から2013年に家禽及び野鳥から分離されたH5及びH7鳥インフルエンザウイルスの遺伝子と抗原性の解析
  大河原 彩子; 日尾野 隆大; 小笠原 浩平; 岡松 正敏; 迫田 義博; 喜田 宏, 日本獣医学会学術集会講演要旨集, 157回, 411, 411, Aug. 2014
  (公社)日本獣医学会, Japanese
• 糖鎖アレイを用いたカモとニワトリのインフルエンザウイルスのレセプター特異性
  岡松 正敏; de Vries Robert; 日尾野 隆大; 迫田 義博; Paulson James C; 喜田 宏, 日本獣医学会学術集会講演要旨集, 157回, 419, 419, Aug. 2014
  (公社)日本獣医学会, Japanese
• ニワトリのインフルエンザウイルスはフコシル化α2,3シアル酸糖鎖をレセプターとする
  日尾野隆大; 岡松正敏; 五十嵐学; 西原祥子; 高瀬明; 迫田義博; 喜田宏, 日本糖質学会年会要旨集, 33rd, 62, 23 Jul. 2014
  Japanese
• H7低病原性鳥インフルエンザウイルスのニワトリに対する病原性獲得メカニズム
  市川貴也; 岡松正敏; 佐藤由佳; 日尾野隆大; 迫田義博; 喜田宏, 日本獣医学会学術集会講演要旨集, 156th, 269, 30 Aug. 2013
  Japanese
• ニワトリのインフルエンザウイルスはニワトリ気管上皮に発現するフコシル化α2,3シアル酸糖鎖をレセプターとして認識する
  日尾野隆大; 岡松正敏; 西原祥子; 高瀬明; 迫田義博; 喜田宏, 日本獣医学会学術集会講演要旨集, 155th, 229, 04 Mar. 2013
  Japanese
• 2010‐11シーズンに国内の野鳥斃死体から分離されたH5N1高病原性鳥インフルエンザウイルスのカモ科およびサギ科鳥類に対する病原性と伝播能の解析
  曽田公輔; 笛吹達史; 宇野有紀子; 永井泰子; 尾崎弘一; 伊藤啓史; 山本直樹; 田村友和; 日尾野隆大; 岡松正敏; 迫田義博; 高田礼人; 村瀬敏之; 山口剛士; 伊藤壽啓, 日本獣医学会学術集会講演要旨集, 155th, 83, 04 Mar. 2013
  Japanese

• 海鳥と海棲哺乳類の高病原性鳥インフルエンザウイルス感染事例におけるワンヘルスアクションとウイルス学的知見
  日尾野隆大; 直亨則; 渡邊有希子; 市川世識; 青島圭佑; 服部薫; 外山雅大; Hew Yik Lim; 磯田典和; 松野啓太; 大沼学; 迫田義博
  第168回日本獣医学会学術集会, 05 Sep. 2025, Invited oral presentation
  03 Sep. 2025 - 04 Sep. 2025, [Invited]
• Multisectoral collaboration against HPAIV cases in marine mammals in Japan
  Takahiro Hiono
  Regional workshop on avian disease prevention and control in Asia and the Pacific 2025, 27 Aug. 2025, Invited oral presentation
  26 Aug. 2025 - 28 Aug. 2025, [Invited]
• Detection and Isolation of H5N1 High Pathogenicity Avian Influenza Viruses from Sea mammals in Association with the Die-off Event of Seabirds in Eastern Hokkaido, Japan
  Takahiro Hiono; Naganori Nao; Yukiko Watanabe; Yoshiki Ichikawa; Kaoru Hattori; Masahiro Toyama; Yik lim Hew; Norikazu Isoda; Keita Matsuno; Manabu Onuma; Yohshihiro Sakoda
  East Asia Wildlife Health Network meeting #16, 25 Jun. 2025, Invited oral presentation
  25 Jun. 2025 - 25 Jun. 2025, [Invited]
• Detection and Isolation of H5N1 High Pathogenicity Avian Influenza Viruses from Sea mammals in Association with the Die-off Event of Seabirds in Eastern Hokkaido, Japan
  Takahiro Hiono; Naganori Nao; Yukiko Watanabe; Yoshiki Ichikawa; Kaoru Hattori; Masahiro Toyama; Yik lim Hew; Norikazu Isoda; Keita Matsuno; Manabu Onuma; Yohshihiro Sakoda
  Asia-Pacific Wildlife Health Workshop 2025, 20 Jun. 2025, Invited oral presentation
  [Invited]
• One Health approach against HPAIVs in wild seabirds and mammals in Hokkaido, Japan
  Takahiro HIONO; Norikazu ISODA; Yik Lim HEW; Yoshihiro SAKODA
  4th Online meeting for the WOAH avian disease network in East Asia, 19 May 2025, Invited oral presentation
  19 May 2025 - 19 May 2025, [Invited]
• Redefining the sialoside receptor distributions in birds and mammals
  Takahiro Hiono; Tatsuru Morita; Daiki Kobayashi; Rio Harada; Norikazu Isoda; Yoshihiro Sakoda
  World Flu Day 2024 Symposium 9th Japan-China Bilateral Symposium on All Influenza Viruses, 02 Nov. 2024, Invited oral presentation
  01 Nov. 2024 - 02 Nov. 2024, [Invited]
• 最近の高病原性鳥インフルエンザウイルスに関する話題提供 (ウシの高病原性鳥インフルエンザウイルス感受性を中心に)
  Takahiro Hiono; Norikazu Isoda; Tatsuru Morita; Yik Lim Hew; Yoshihiro Sakoda
  第37回インフルエンザウイルス研究者交流の会シンポジウム, 04 Jul. 2024, Invited oral presentation
  04 Jul. 2024 - 06 Jul. 2024, [Invited]
• Cocirculation of two genetically distinct high pathogenicity avian influenza viruses in Hokkaido, Japan
  Takahiro HIONO; Norikazu ISODA; Yik Lim HEW; Yoshihiro SAKODA
  3rd Online meeting for the WOAH avian disease network in East Asia, 02 Apr. 2024, Invited oral presentation
  02 Apr. 2024 - 02 Apr. 2024, [Invited]
• 高病原性鳥インフルエンザウイルスの最新事情とOne Health
  日尾野 隆大; 磯田 典和; 迫田 義博
  埼玉大学先端産業国際ラボラトリー メディカルイノベーション研究ユニット 第26回ワークショップ 感染症に挑むII, 20 Dec. 2023, Invited oral presentation
  20 Dec. 2023 - 20 Dec. 2023, [Invited]
• 高病原性鳥インフルエンザウイルスの現状とOne Health
  日尾野 隆大; 磯田 典和; 迫田 義博
  第 23 回 人と動物の共通感染症研究会学術集会, 28 Oct. 2023, Invited oral presentation
  28 Oct. 2023 - 28 Oct. 2023, [Invited]
• Glycoscientific approaches toward understanding the host preference of avian influenza viruses
  Takahiro Hiono
  The 6th One Health Lecture Series on Solutions to Global One Health Challenges Towards Human Animals and Environmental Health, 10 Dec. 2021, English, Invited oral presentation
  10 Dec. 2021 - 11 Dec. 2021, [Invited]
• 鳥インフルエンザウイルスの糖結合特異性
  日尾野隆大
  第31回 インフルエンザ研究者交流の会シンポジウム, 08 Jun. 2017, Invited oral presentation
  08 Jun. 2017 - 10 Jun. 2017, [Invited]

• 産業動物獣医療実習Ⅱ, 2024年, 学士課程, 獣医学部
• 産業動物獣医療実習Ⅱ, 2024年, 学士課程, 獣医学部
• 微生物学実習, 2024年, 学士課程, 獣医学部
• 動物衛生学実習, 2024年, 学士課程, 獣医学部
• アドバンスト演習, 2024年, 学士課程, 獣医学部
• 草地飼料学, 2024年, 学士課程, 獣医学部

• THE JAPANESE SOCIETY OF CARBOHYDRATE RESEARCH
• THE JAPANESE SOCIETY OF VETERINARY SCIENCE
• THE JAPANESE SOCIETY FOR VIROLOGY

• Glycoforms of viral proteins and their immunogenicity
  Grants-in-Aid for Scientific Research
  01 Apr. 2024 - 31 Mar. 2027
  日尾野 隆大
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, 24K09260
• 宿主RNAメチルトランスフェラーゼを標的とした抗インフルエンザウイルス薬の探索
  科学研究費助成事業 国際共同研究加速基金(国際共同研究強化(B))
  07 Oct. 2022 - 31 Mar. 2026
  五十嵐 学; 松野 啓太; 日尾野 隆大
  日本学術振興会, 国際共同研究加速基金(国際共同研究強化(B)), 北海道大学, 22KK0094
• ウイルスに付加する糖鎖の異種宿主間病原体伝播における新規生物学的機能の解明
  科学研究費助成事業 若手研究
  01 Apr. 2021 - 31 Mar. 2024
  日尾野 隆大
  日本学術振興会, 若手研究, 北海道大学, 21K14982
• ウイルス感染における糖鎖リモデリングの意義解明に向けた技術基盤の構築
  Grant-in-Aid for Early-Career Scientists
  2018 - 2020
  Takahiro Hiono
  Japan Society for the Promotion of Science, Principal investigator, Competitive research funding
• Comprehensive analyses of the interaction between host glycan and influenza viruses and its significance in virus infection
  Grant-in-Aid for Research Activity start-up
  2016 - 2017
  Takahiro Hiono
  インフルエンザウイルスは糖と密接に関連した病原体である。本研究では、インフルエンザウイルスと糖質の直接的な相互作用に加えて、感染に対する宿主免疫応答としての糖質の発現変動を解析することで、その宿主域と病原性を規定する因子の同定を試みた。インフルエンザウイルスのHAの当結合特異性およびNAの基質特異性を解析する手法を確立した。またインフルエンザウイルス感染細胞では様々な糖鎖変化が認められることをレクチンマイクロアレイで明らかにした。
  Japan Society for the Promotion of Science, Principal investigator, Competitive research funding
• インフルエンザウイルス感染におけるレセプターとしてのシアル酸糖蛋白質の機能解析
  科学研究費助成事業
  24 Apr. 2015 - 31 Mar. 2016
  日尾野 隆大
  バイオインフォマティクスの手法を用いて、ニワトリから分離されたH5亜型鳥インフルエンザウイルスのヘマグルチニン (HA)とニワトリ型レセプターであるフコシル化α2,3シアロ糖の結合を予測した。その結果、フコシル化α2,3シアロ糖のフコース基はH5HA分子の222および227位のアミノ酸と相互作用することが予測された。
  カモから分離されたA/duck/Mongolia/54/2001 (H5N2) (Dk/MNG)株およびニワトリから分離されたA/chicken/Ibaraki/1/2005 (H5N2) (Ck/IBR)株の組換えHAを作出し、その糖鎖結合特異性をGlycan microarray法で評価した。その結果、Dk/MNG株のHAはフコシル化のないα2,3シアロ糖と、Ck/IBRのHAはフコシル化α2,3シアロ糖とそれぞれ特異的に結合することが分かった。さらに、これらの糖鎖結合特異性が、HAの222位および227位のアミノ酸モチーフによって規定されていることを明らかにした。一方で、Dk/MNG株の粒子はフコシル化α2,3シアロ糖とフコシル化のないα2,3シアロ糖の両方と結合した。この結果はフコシル化のないα2,3シアロ糖とのみ結合した組換えHAの成績と異なっていた。そこでノイラミニダーゼ(NA)阻害剤の存在下で同様の試験を行ったところ、Dk/MNG株の粒子はフコシル化のないα2,3シアロ糖のみと特異的に結合した。すなわち、Dk/MNG株の粒子はNAを介してフコシル化α2,3シアロ糖を認識していると考えられる。
  また、インフルエンザウイルスが細胞内に侵入する際に利用する宿主のエンドサイトーシス経路を明らかにするために、クラスリン依存性エンドサイトーシスのアダプター蛋白質であるEPN1およびAP2M1のノックダウンを試みた。当初、shRNA発現ベクターを用いたノックダウンを計画していたが、RNA干渉の効果を得ることができなかった。そこで合成siRNAを用いてノックダウンを試みたところ、EPN1およびAP2M1共に概ね70%以上のノックダウンに成功した。
  日本学術振興会, 特別研究員奨励費, 北海道大学, 15J00209