SEKIGUCHI Satoshi

写真a

Affiliation

Center for Animal Disease Control

Title

Professor

External Link

Degree 【 display / non-display

  • Doctor (Veterinary Medical Sciences) ( 2007.3   The University of Tokyo )

Research Interests 【 display / non-display

  • Veterinary epidemiology

Research Areas 【 display / non-display

  • Life Science / Veterinary medical science

Education 【 display / non-display

  • The University of Tokyo   Graduate School of Agricultural and Life Sciences   Veterinary Medical Sciences

    - 2007.3

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    Country:Japan

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  • Nippon Veterinary and Animal Science University   Faculty of Veterinary Medicine   Department of Veterinary Science

    - 2003.3

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    Country:Japan

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Campus Career 【 display / non-display

  • University of Miyazaki   Center for Animal Disease Control   Professor

    2023.08 - Now

  • University of Miyazaki   Center for Animal Disease Control   Division of Prevention & Control for Animal Diseases   Associate Professor

    2015.10 - 2023.07

  • University of Miyazaki   Faculty of Agriculture   Department of Veterinary Science   Animal infectious disease and prevention study course   Associate Professor

    2012.03 - 2023.07

External Career 【 display / non-display

  • 宮崎大学 産業動物防疫リサーチセンター   防疫戦略部門   部門長

    2022.4

  • University of Miyazaki   Center for Animal Disease Control Division of Prevention & Control for Animal Diseases   Associate Professor

    2015.10

  • University of Miyazaki   Faculty of Agriculture Department of Veterinary Science Animal infectious disease and prevention study course   Associate Professor

    2012.3

  • The University of Tokyo   Researcher

    2010.8 - 2012.2

  • Tokyo Metropolitan Institute of Medical Science   Researcher

    2007.4 - 2010.7

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Professional Memberships 【 display / non-display

  • Frontiers Veterinary Science

    2020.4

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  • 鶏肉の食中毒菌低減を進めるための検討会

    2020.4

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  • 獣医事審議会専門委員会

    2018.4 - 2021.3

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  • Japan Veterinary Medical Association

    2012.10

  • Japanese Society of Veterinary Science

    2012.8

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Papers 【 display / non-display

  • Development of droplet digital PCR for quantification of bovine leukemia virus proviral load using unpurified genomic DNA Reviewed International coauthorship

    Wu X., Notsu K., Matsuura Y., Mitoma S., El Daous H., Norimine J., Sekiguchi S.

    Journal of Virological Methods   315   114706   2023.5

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    Authorship:Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Journal of Virological Methods  

    Bovine leukemia virus (BLV) is the causative agent of a B-cell tumor called enzootic bovine leukosis. Preventing BLV spreading is required to reduce economic loss related to BLV infection of livestock. To quantify proviral load (PVL) more easily and rapidly, we developed a quantification system of PVL using droplet digital PCR (ddPCR). This method uses a multiplex TaqMan assay of the BLV provirus and housekeeping gene RPP30 for the quantification of BLV in BLV-infected cells. Furthermore, we combined ddPCR with DNA purification-free sample preparation (unpurified genomic DNA). The percentage of BLV-infected cells based on unpurified genomic DNA was highly correlated with that based on purified genomic DNA (correlation coefficient: 0.906). Thus, this new technique is a suitable method to quantify PVL of BLV-infected cattle in a large sample number.

    DOI: 10.1016/j.jviromet.2023.114706

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    PubMed

  • Identifying Pathogen and Allele Type Simultaneously in a Single Well Using Droplet Digital PCR. Reviewed International coauthorship

    Notsu K, El Daous H, Mitoma S, Wu X, Norimine J, Sekiguchi S

    mSphere   8 ( 1 )   e0049322   2023.1

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    Authorship:Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:mSphere  

    In the transmission control of chronic and untreatable livestock diseases such as bovine leukemia virus (BLV) infection, the removal of viral superspreaders is a fundamental approach. On the other hand, selective breeding of cattle with BLV-resistant capacity is also critical for reducing the viral damage to productivity by keeping infected cattle. To provide a way of measuring BLV proviral load (PVL) and identifying susceptible/resistant cattle simply and rapidly, we developed a fourplex droplet digital PCR method targeting the BLV pol gene, BLV-susceptible bovine major histocompatibility complex (BoLA)-DRB3*016:01 allele, resistant DRB3*009:02 allele, and housekeeping RPP30 gene (IPATS-BLV). IPATS-BLV successfully measured the percentage of BLV-infected cells and determined allele types precisely. Furthermore, it discriminated homozygous from heterozygous carriers. Using this method to determine the impact of carrying these alleles on the BLV PVL, we found DRB3*009:02-carrying cattle could suppress the PVL to a low or undetectable level, even with the presence of a susceptible heterozygous allele. Although the population of DRB3*016:01-carrying cattle showed significantly higher PVLs compared with cattle carrying other alleles, their individual PVLs were highly variable. Because of the simplicity and speed of this single-well assay, our method has the potential of being a suitable platform for the combined diagnosis of pathogen level and host biomarkers in other infectious diseases satisfying the two following characteristics of disease outcomes: (i) pathogen level acts as a critical maker of disease progression; and (ii) impactful disease-related host genetic biomarkers are already identified. IMPORTANCE While pathogen-level quantification is an important diagnostic of disease severity and transmissibility, disease-related host biomarkers are also useful in predicting outcomes in infectious diseases. In this study, we demonstrate that combined proviral load (PVL) and host biomarker diagnostics can be used to detect bovine leukemia virus (BLV) infection, which has a negative economic impact on the cattle industry. We developed a fourplex droplet digital PCR assay for PVL of BLV and susceptible and resistant host genes named IPATS-BLV. IPATS-BLV has inherent merits in measuring PVL and identifying susceptible and resistant cattle with superior simplicity and speed because of a single-well assay. Our new laboratory technique contributes to strengthening risk-based herd management used to control within-herd BLV transmission. Furthermore, this assay design potentially improves the diagnostics of other infectious diseases by combining the pathogen level and disease-related host genetic biomarker to predict disease outcomes.

    DOI: 10.1128/msphere.00493-22

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  • A pooled testing system to rapidly identify cattle carrying the elite controller BoLA-DRB3*009:02 haplotype against bovine leukemia virus infection. Reviewed International coauthorship

    Notsu K, Daous HE, Mitoma S, Norimine J, Sekiguchi S

    HLA   99 ( 1 )   12 - 24   2021.11

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    Authorship:Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:HLA  

    As genetically resistant individuals, the “elite controllers” (ECs) of human immunodeficiency virus infection have been focused on as the keys to developing further functional treatments in medicine. In the livestock production field, identifying the ECs of bovine leukemia virus (BLV) infection in cattle is desired to stop BLV transmission chains on farms. Cattle carrying the bovine leukocyte antigen (BoLA)-DRB3*009:02 allele (DRB3*009:02) have a strong possibility of being BLV ECs. Most of cattle carrying this allele maintain undetectable BLV proviral loads and do not shed virus even when infected. BLV ECs can act as transmission barriers when placed between uninfected and infected cattle in a barn. To identify cattle carrying DRB3*009:02 in large populations more easily, we developed a pooled testing system. It employs a highly sensitive, specific real-time PCR assay and TaqMan MGB probes (DRB3*009:02-TaqMan assay). Using this system, we determined the percentage of DRB3*009:02-carrying cattle on Kyushu Island, Japan. Our pooled testing system detected cattle carrying the DRB3*009:02 allele from a DNA pool containing one DRB3*009:02-positive animal and 29 cattle with other alleles. Its capacity is sufficient for herd-level screening for DRB3*009:02-carrying cattle. The DRB3*009:02-TaqMan assay showed high-discriminative sensitivity and specificity toward DRB3*009:02, making it suitable for identifying DRB3*009:02-carrying cattle in post-screening tests on individuals. We determined that the percentage of DRB3*009:02-carrying cattle in Kyushu Island was 10.56%. With its ease of use and reliable detection, this new method strengthens the laboratory typing for DRB3*009:02-carrying cattle. Thus, our findings support the use of BLV ECs in the field.

    DOI: 10.1111/tan.14502

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  • Relationship between allelic heterozygosity in bola-drb3 and proviral loads in bovine leukemia virus-infected cattle Reviewed International coauthorship

    Daous H.E., Mitoma S., Elhanafy E., Huyen N.T., Ngan M.T., Notsu K., Kaneko C., Norimine J., Sekiguchi S.

    Animals   11 ( 3 )   1 - 14   2021.3

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    Authorship:Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Animals  

    Enzootic bovine leukosis is a lethal neoplastic disease caused by bovine leukemia virus (BLV), belongs to family Retroviridae. The BLV proviral load (PVL) represents the quantity of BLV genome that has integrated into the host’s genome in BLV-infected cells. Bovine leukocyte antigen (BoLA) class II allelic polymorphisms are associated with PVLs in BLV-infected cattle. We sought to identify relationships between BoLA-DRB3 allelic heterozygosity and BLV PVLs among different cattle breeds. Blood samples from 598 BLV-infected cattle were quantified to determine their PVLs by real-time polymerase chain reaction. The results were confirmed by a BLV-enzyme-linked immunosorbent assay. Restriction fragment length polymorphism-polymerase chain reaction identified 22 BoLA-DRB3 alleles. Multivariate negative binomial regression modeling was used to test for associations between BLV PVLs and BoLA-DRB3 alleles. BoLA-DRB3.2*3, *7, *8, *11, *22, *24, and *28 alleles were significantly associated with low PVLs. BoLA-DRB3.2*10 was significantly associated with high PVLs. Some heterozygous allele combinations were associated with low PVLs (*3/*28, *7/*8, *8/*11, *10/*11, and *11/*16); others were associated with high PVLs (*1/*41, *10/*16, *10/*41, *16/*27, and *22/*27). Interestingly, the BoLA-DRB3.2*11 heterozygous allele was always strongly and independently associated with low PVLs. This is the first reported evidence of an association between heterozygous allelic combinations and BLV PVLs.

    DOI: 10.3390/ani11030647

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  • The detection of long-lasting memory foot-and-mouth disease (FMD) virus serotype O-specific CD4<sup>+</sup> T cells from FMD-vaccinated cattle by bovine major histocompatibility complex class II tetramer Reviewed International coauthorship

    Mitoma S., Carr B.V., Harvey Y., Moffat K., Sekiguchi S., Charleston B., Norimine J., Seago J.

    Immunology   164 ( 2 )   266 - 278   2021

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Immunology  

    Foot-and-mouth disease (FMD) is a highly contagious, economically devastating disease of cloven-hooved animals. The development of long-lasting effective FMD vaccines would greatly benefit the global FMD control programme. Deep analysis of adaptive immunity in cattle vaccinated against FMD is technically challenging due to the lack of species-specific tools. In this study, we aimed to identify CD4+ T-cell epitopes in the FMD virus (FMDV) capsid and to phenotype the CD4+ T cells that recognize them using bovine major histocompatibility complex (BoLA) class II tetramer. A BoLA class II tetramer based on the DRA/DRB3*020:02 allele and FMDV antigen-stimulated PBMCs from bovine vaccinates were used to successfully identify four epitopes in the FMDV capsid, three of which have not been previously reported; two epitopes were identified in the structural protein VP1, one in VP3 and one in VP4. Specificity of the three novel epitopes was confirmed by proliferation assay. All epitope-expanded T-cell populations produced IFN-γ in vitro, indicating a long-lasting Th1 cell phenotype after FMD vaccination. VP3-specific CD4+ T cells exhibited the highest frequency amongst the identified epitopes, comprising >0·004% of the CD4+ T-cell population. CD45RO+CCR7+ defined central memory CD4+ T-cell subpopulations were present in higher frequency in FMDV-specific CD4+ T-cell populations from FMD-vaccinated cattle ex vivo. This indicates an important role in maintaining cell adaptive immunity after FMD vaccination. Notably, FMDV epitope-loaded tetramers detected the presence of FMDV-specific CD4+ T cells in bovine PBMC more than four years after vaccination. This work contributes to our understanding of vaccine efficacy.

    DOI: 10.1111/imm.13367

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Books 【 display / non-display

  • veterinary epidemiology

    ( Role: Contributor)

    2022.3 

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    Language:Japanese Book type:Textbook, survey, introduction

  • 動物衛生学

    髙井伸二,末吉益男,永幡肇,他( Role: Joint author)

    文永堂出版  2018.4 

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    Total pages:415   Responsible for pages:125-136   Language:Japanese Book type:Textbook, survey, introduction

MISC 【 display / non-display

  • 牛伝染性リンパ腫の新しい検査法とその利用 Invited

    関口 敏

    畜産技術   ( 816 )   36 - 40   2023.5

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    Authorship:Lead author, Corresponding author   Language:Japanese   Publishing type:Article, review, commentary, editorial, etc. (scientific journal)  

  • Corrigendum to “Development of droplet digital PCR for quantification of bovine leukemia virus proviral load using unpurified genomic DNA” [J. Virol. Methods, 315 (2023) 114706] (Journal of Virological Methods (2023) 315, (S0166093423000319), (10.1016/j.jviromet.2023.114706))

    Wu X., Notsu K., Matsuura Y., Mitoma S., El Daous H., Norimine J., Sekiguchi S.

    Journal of Virological Methods   315   114708   2023.5

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    Language:Japanese   Publishing type:Rapid communication, short report, research note, etc. (scientific journal)   Publisher:Journal of Virological Methods  

    The authors regret that several typing errors in paragraph of 2.7. Validation of ddPCR for amplicon-containing plasmid DNA. The correct formula is provided below: [Formula presented] Furthermore, Figure Supplement 1 contained a few editorial errors, the correct version is included below. The authors would like to apologise for any inconvenience caused.

    DOI: 10.1016/j.jviromet.2023.114708

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  • 地域とともに実現した牛伝染性リンパ腫の持続可能な防疫対策 Invited

    関口 敏

    家畜感染症学会誌   2022.9

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    Authorship:Lead author, Corresponding author   Language:Japanese   Publishing type:Article, review, commentary, editorial, etc. (scientific journal)  

  • Editorial: The Epidemiology, Diagnosis and Prevention of Infectious Diseases in Livestock Invited International coauthorship

    Sekiguchi S., Wiratsudakul A., Nguyen V.G.

    Frontiers in Veterinary Science   8   840635   2022.1

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    Authorship:Lead author   Language:English   Publishing type:Article, review, commentary, editorial, etc. (scientific journal)   Publisher:Frontiers in Veterinary Science  

    DOI: 10.3389/fvets.2021.840635

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  • Quantitative Risk Assessment of Introduction of Bovine Leukemia Virus in Beef Herds Using Deterministic Method

    藤原未歩, 牛谷雄一, 野津昂亮, EL-DAOUS Hala, 芹田光玲, 三苫修也, 乗峰潤三, 乗峰潤三, 関口敏, 関口敏

    獣医疫学雑誌   25 ( 1 )   2021

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    Publishing type:Rapid communication, short report, research note, etc. (scientific journal)  

    J-GLOBAL

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Presentations 【 display / non-display

  • 確率論的手法を用いた肉用牛の外部導入における 牛伝染性リンパ腫ウイルス感染のリスク評価 International coauthorship

    藤原未歩、牛谷雄一、野津昴亮、Hala El-Daous、芹田光玲、 三苫修也、乗峰潤三、関口敏

    第164回日本獣医学会 

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    Event date: 2021.9.7 - 2021.9.13

    Language:Japanese   Presentation type:Oral presentation (general)  

  • CADIC生物統計学講座

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    CADICリスク分析学 

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    Event date: 2022.3.22 - 2022.3.23

    Language:Japanese   Presentation type:Oral presentation (general)  

  • A pooled testing system to rapidly identify cattle carrying the elite controller BoLA-DRB3*009:02 haplotype against bovine leukemia virus infection Invited International coauthorship

    関口 敏

    宮崎大学若手研究者論文発表支援報告会 

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    Event date: 2022.3.14

    Language:Japanese   Presentation type:Public lecture, seminar, tutorial, course, or other speech  

  • SATREPS JCC meeting International coauthorship International conference

    関口 敏

    SATREPS JCC meeting 

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    Event date: 2022.3.11

    Language:English   Presentation type:Oral presentation (general)  

  • 90分でわかる牛ウイルス性下痢症 Invited

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    令和3年度家畜生産農場衛生対策事業​ 

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    Event date: 2022.2.18

    Language:Japanese   Presentation type:Oral presentation (invited, special)  

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Awards 【 display / non-display

  • 令和5年度文部科学大臣表彰 科学技術賞(理解増進部門)

    2023.4   文部科学省  

    関口 敏

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    Award type:International academic award (Japan or overseas) 

  • Top Downloaded Article (HLA: IMMUNE RESPONSE GENETICS)

    2023.4   WILEY   A pooled testing system to rapidly identify cattle carrying the elite controllerBoLA-DRB3*009:02 haplotype against bovine leukemia virus infection

    Kosuke Notsu, Hala El Daous, Shuya Mitoma, Junzo Norimine, Satoshi Sekiguchi

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    Award type:Honored in official journal of a scientific society, scientific journal 

  • 2022年宮崎大学農学部地域貢献優秀賞

    2022.3   宮崎大学  

    関口 敏

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    Award type:International academic award (Japan or overseas) 

  • 宮崎大学若手研究者論文発表支援

    2022.3   宮崎大学  

    関口敏

  • Japan Award

    2021.11  

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    Award type:International academic award (Japan or overseas) 

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Grant-in-Aid for Scientific Research 【 display / non-display

  • 牛白血病ウイルス抵抗性牛の簡易判別診断法の開発

    Grant number:22KJ2524   2021.04 - 2024.03

    独立行政法人日本学術振興会  科学研究基金  特別研究員奨励費

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    Authorship:Principal investigator 

  • 牛白血病ウイルスの抵抗性牛の簡易判別診断法の開発

    Grant number:21J23396  2021.04 - 2024.03

    独立行政法人日本学術振興会  科学研究費補助金  特別研究員奨励費

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    Authorship:Principal investigator 

  • 牛白血病ウイルスの血中プロウイルス量と感染細胞数に関する研究

    Grant number:20K06413  2020.04 - 2024.03

    独立行政法人日本学術振興会  科学研究費補助金  基盤研究(C)

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    Authorship:Principal investigator 

    牛白血病ウイルス(BLV)は牛のリンパ球(B細胞)に感染し,宿主のDNA中にプロウイルスとして組み込まれる。そのため血中プロウイルス量は,感染細胞数に比例するといわれ,感染力を評価する指標として知られている。しかし,血中のプロウイルス量だけでは感染力を説明できない個体も少なくない。そこで本研究では,BLVの感染力を説明する「第2の関連因子」を同定することを目的とする。まず,感染力には感染細胞あたりのプロウイルスの挿入個数が影響しているという仮説の下,プロウイルスを高精度に測定する絶対定量法を確立する。次に,臨床検体を用いて血中プロウイルス量とプロウイルスの挿入個数の関係を明らかにする。最後に,プロウイルスの挿入個数が感染力に及ぼす影響をフィールド調査で分析する。本研究によって新たな関連因子が同定されれば,より正確に感染リスクを評価することが可能となる。

  • 豚流行性下痢の感染拡大要因の解明

    2015.04 - 2018.03

    科学研究費補助金  若手研究(B)

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    Authorship:Principal investigator 

    本年(2014年)、全国的に膨大な経済的被害を出した豚流行性下痢(PED)は未だに感染が拡大している状況にある。その一方で、発生農家に近接していてもウイルスの侵入を防いでいる非感染農場が存在することも周知の事実である。このことから、感染農場と非感染農場の飼養管理方法や防疫対策方法を比較すれば、ウイルスの感染拡大に関与するリスク因子と防御因子を同定することが可能と予測される。本研究では、能動的サーベイランスを用いてPEDウイルスの感染農場と非感染農場を特定し、その相違点を明らかにすることでPEDの感染拡大要因の解明を行う(具体的技術については計画と方法に記載)。本研究によって、PEDの感染拡大する要因が解明されれば、科学的根拠に基づいた効果的な防疫対策の立案が可能になると期待される。

  • 免疫系の攪乱によるC型肝炎ウイルスの持続感染化及び発症機序の解析とその制御

    2009.04 - 2011.03

    科学研究費補助金  特定領域研究

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    Authorship:Coinvestigator(s) 

    【目的】C型肝炎は国内で約200万人に及ぶ感染者が存在し、安全で効果的な治療法の開発が急務となっている。我々はHCV遺伝子組換えワクチニアウイルス(HCV-RVV)株を樹立し、HCV持続感染モデルマウスを用いて治療ワクチンとしての有用性を検討した。

    【方法】Cre/loxPシステムでHCV遺伝子を導入したTg(Cre/loxP/HCV-Tg)マウスと、IFN誘導性のCreを発現するTgマウスを交配させ、任意の時期にHCV遺伝子をスイッチング発現するTgマウス(Cre/loxP/HCV-MxCreTg)を作製した。HCV-RVVはHCVの構造蛋白質を主に発現するCN2、非構造蛋白質を発現するN25、全蛋白質を発現するCN5を用いた。これらHCV-RVVの治療効果を評価するために、HCV蛋白を持続的に発現した状態のTgマウスに単回皮内接種し、接種後1週および4週のマウスにおいて解析を行った。

    【成績】HCV-RVV接種後1週のTgマウス肝臓では各接種群でHCV蛋白の減少が認められなかったが、4週ではN25接種群が減少していた。またN25接種群では接種後1週で肝臓の索状構造や肝細胞のsteatosisなど、HCV特有の形態学的な異常の正常化が認められた。N25接種群においてTgマウスのCD4またはCD8を欠損させるとHCV蛋白減少効果はみられなかったが、HCV特有の形態学的な異常の改善が認められた。

    【結論】我々が樹立したTgマウスは慢性肝炎を発症させることができるHCV持続感染モデルは、HCVの持続感染化と宿主の免疫応答の関係を解析することを可能にした。さらに、HCV-RVV(N25)接種によりマウス肝臓におけるHCV蛋白の減少及び慢性肝炎症状の正常化が認められた。HCVの肝細胞からの除去にはCD4,CD8 T細胞が関与しているが、HCVによる肝細胞の形態学的異常はHCV蛋白の発現に依存するのではなく他のメカニズムが関与していることが示唆された。

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Other external funds procured 【 display / non-display

  • Risk analysis of antimicrobial resistance transmission at the wildlife-livestock-human interface in Zambia

    2017.04 - 2020.03

    JSPS RONPAKU Program 

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    Grant type:Competitive

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Committee Memberships 【 display / non-display

  • BVD対策推進コンソーシアム   BVD対策推進コンソーシアム会員  

    2023.4   

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    Committee type:学協会

  • vetCBT問題作成・精選委員会   vetCBT問題作成・精選委員  

    2022.4   

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    Committee type:学協会

  • 日本獣医学会   編集委員  

    2021.4   

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    Committee type:学協会

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  • 鶏肉の食中毒菌低減を進めるための検討会   検討委員  

    2020.4   

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    Committee type:政府

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  • Frontiers Veterinary Science   Topic editor  

    2020.4 - 2022.3   

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    Committee type:学協会

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Social Activities 【 display / non-display

  • 家畜衛生検査受託研究事業

    Role(s): Advisor, Informant, Planner, Organizing member

    2020.4.1 - Now

  • 清武町西新町青少年育成委員会委員長

    Role(s): Planner, Organizing member, Demonstrator

    2022.4.1 - Now

  • 牛伝染性リンパ腫の疫学調査

    Role(s): Investigater

    2022.4.1 - 2023.3.31

  • 地域との協働による高等学校教育改革推進事業

    Role(s): Lecturer

    2022.4.1 - 2023.3.31

  • BLVパンフレットの作成

    Role(s): Chief editor, Editer, Advisor, Informant, Planner

    2022.3.24 - Now

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Media Coverage 【 display / non-display

  • 牛リンパ腫抵抗性判別 Newspaper, magazine

    日本農業新聞  2023.3.8

  • 宮大 格安検査準備進む Newspaper, magazine

    宮崎日日新聞  2022.10.24

  • DX(デジタルトランスフォーメーション)に関する取組事例

    2022.1.13

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    Author:Myself 

  • 獣医医事審議会専門員

Academic Activities 【 display / non-display

  • 牛伝染性リンパ腫の検査および防疫対策に関するコンサルティング活動

    2013.4.1 - Now

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    Type:Academic research 

  • 牛ウイルス性下痢症の検査および防疫対策に関するコンサルティング活動

    Role(s): Planning/Implementing academic research

    2013.4.1 - Now

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    Type:Academic research