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Affiliation |
Center for Animal Disease Control |
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Professor |
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External Link |
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SEKIGUCHI Satoshi
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Degree 【 display / non-display 】
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Doctor (Veterinary Medical Sciences) ( 2007.3 The University of Tokyo )
Research Areas 【 display / non-display 】
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Life Science / Veterinary medical science
Education 【 display / non-display 】
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The University of Tokyo Graduate School of Agricultural and Life Sciences Veterinary Medical Sciences
- 2007.3
Country：Japan
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Nippon Veterinary and Animal Science University Faculty of Veterinary Medicine Department of Veterinary Science
- 2003.3
Country：Japan
Campus Career 【 display / non-display 】
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University of Miyazaki Center for Animal Disease Control Professor
2023.08 - Now
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University of Miyazaki Center for Animal Disease Control Division of Prevention & Control for Animal Diseases Associate Professor
2015.10 - 2023.07
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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 】
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The University of Tokyo Researcher
2010.8 - 2012.2
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Tokyo Metropolitan Institute of Medical Science Researcher
2007.4 - 2010.7
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The University of Tokyo Special researcher of the Japan Society for the Promotion of Science
2005.4 - 2007.3
Professional Memberships 【 display / non-display 】
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Japanese Society of Veterinary Science
2012.8
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The Japan Society of Veterinary Epidemiology
2012.8
Papers 【 display / non-display 】
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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
Authorship：Last author, 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.
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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
Authorship：Last author, 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.
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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
Authorship：Last author, 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
Authorship：Last author, 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
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
Books 【 display / non-display 】
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veterinary epidemiology
（ Role： Contributor）
2022.3
Language：Japanese Book type：Textbook, survey, introduction
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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
Total pages：415 Responsible for pages：125-136 Language：Japanese Book type：Textbook, survey, introduction
MISC 【 display / non-display 】
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牛ウイルス性下痢（BVD）対策の経済的評価 Invited
関口 敏
臨床獣医 42 ( 4 ) 40 - 44 2024.4
Authorship：Lead author Language：Japanese Publishing type：Article, review, commentary, editorial, etc. (trade magazine, newspaper, online media)
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家畜伝染病対策に関する検査技術の開発および社会実装 Invited
関口 敏
調査月報 369 2 - 7 2024.2
Authorship：Lead author Language：Japanese Publishing type：Article, review, commentary, editorial, etc. (trade magazine, newspaper, online media)
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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
Authorship：Lead author, Corresponding author Language：Japanese Publishing type：Article, review, commentary, editorial, etc. (scientific journal)
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Wu X., Notsu K., Matsuura Y., Mitoma S., El Daous H., Norimine J., Sekiguchi S.
Journal of Virological Methods 315 114708 2023.5
Language：English 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.
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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
Authorship：Lead author, Corresponding author Language：Japanese Publishing type：Article, review, commentary, editorial, etc. (scientific journal)
Presentations 【 display / non-display 】
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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
Event date： 2021.9.7 - 2021.9.13
Language：Japanese Presentation type：Oral presentation (general)
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乾燥血液を用いた牛伝染性リンパ腫ウイルスのプロウイルス量の 定量 International coauthorship
廣川 万莉 、野津 昂亮 、鄔 心悦 、乗峰 潤三 、関口 敏
第63回獣医疫学会学術集会 2024.3.17
Event date： 2024.3.17
Language：Japanese Presentation type：Oral presentation (general)
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宮崎県における牛ウイルス性下痢の積極的疫学調査と分離株の遺伝 子 系統 解析 Invited International coauthorship
野津昂亮 、 龍千香 、 Luqman Ul Hakim Bin Rahmat、 Shakir Ullah、 植木萌 葉 、 三苫修也 、 乗峰潤三 、 青木博史 、亀山健一郎 、迫田義 博 、 関口敏
第63回獣医疫学会学術集会 2024.3.17
Event date： 2024.3.17
Language：Japanese Presentation type：Oral presentation (general)
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牛伝染性リンパ腫ウイルスの検査技術の開発と社会実装 Invited
関口 敏
宮崎大学産業動物防疫リサーチセンターと大分大学グローカル感染症研究センターとの共同セミナー 2023.11.27
Event date： 2023.11.27
Language：Japanese Presentation type：Public lecture, seminar, tutorial, course, or other speech
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牛ウイルス性下痢 制圧に向けた対策 Invited
関口 敏
2023年度九州沖縄産業動物臨床研究会・牛臨床寄生虫研究会合同研究集会 2023.11.25
Event date： 2023.11.25
Language：Japanese Presentation type：Public lecture, seminar, tutorial, course, or other speech
Awards 【 display / non-display 】
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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
Award type：International academic award (Japan or overseas)
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若手優秀発表賞
2024.3 獣医疫学会 宮崎県における牛ウイルス性下痢の積極的疫学調査と分離株の遺伝子系統解析
野津昂亮、龍千香、Luqman Ul Hakim Bin Rahmat、Shakir Ullah、植木萌葉、三苫修也、乗峰潤三、青木博史、亀山健一郎、迫田義博、関口敏
Award type：Award from Japanese society, conference, symposium, etc.
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学生優秀発表賞
2024.3 獣医疫学会 乾燥血液を用いた牛伝染性リンパ腫ウイルスのプロウイルス量の定量
廣川 万莉、野津 昂亮、鄔 心悦、乗峰 潤三、関口 敏
Award type：Award from Japanese society, conference, symposium, etc.
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最優秀学術賞
2023.12 家畜感染症学会 牛伝染性リンパ腫ウイルス抵抗性マーカー遺伝子の簡易検査法の開発
野津昂亮、関口敏
Award type：Award from Japanese society, conference, symposium, etc.
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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
Award type：Honored in official journal of a scientific society, scientific journal
Grant-in-Aid for Scientific Research 【 display / non-display 】
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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
Authorship：Principal investigator
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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
Authorship：Principal investigator
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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)
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。
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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)
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。
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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
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。
Other external funds procured 【 display / non-display 】
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Risk analysis of antimicrobial resistance transmission at the wildlife-livestock-human interface in Zambia
2017.04 - 2020.03
JSPS RONPAKU Program
Grant type：Competitive
Available Technology 【 display / non-display 】
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地域における家畜伝染病のコントロールに関する研究
家畜伝染病の検査法の開発
最適な検査プログラムの開発Home Page： 産業動物伝染病防疫学研究室
Related fields where technical consultation is available：・家畜伝染病の疫学調査、検査方法およびその対策方法に関する相談
・統計解析手法に関するコンサルティングおよび講習Message：動物を病気にさせないことは、動物の命を守るだけでなく、私たちの食の安全・安心を守ることにもつながります。地域が一体となって感染症対策に取り組みましょう。
Committee Memberships 【 display / non-display 】
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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
Committee type：学協会
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vetCBT問題作成・精選委員会 vetCBT問題作成・精選委員
2022.4
Committee type：学協会
Social Activities 【 display / non-display 】
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牛伝染性リンパ腫と牛ウイルス性下痢の病態と防疫対策
Role(s)： Lecturer
令和5年度西部家畜保健衛生推進協議会 畜産講演会 2024.3.8
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牛個体識別AIを起点とする飼養衛生管理と 防疫対策のDX化
Role(s)： Lecturer
都城市 飼育衛生管理と情報通信の融合 2024.3.4
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牛伝染性リンパ腫対策と今後の展望について
Role(s)： Lecturer
小林市 小林市畜産振興大会2024 2024.2.16
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牛伝染性リンパ腫対策に役立つ検査技術について
Role(s)： Lecturer
静岡県 令和５年度公益社団法人静岡県獣医師会産業動物部会講演会 2024.2.9
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牛ウイルス性下痢の現状と対策ついて
Role(s)： Lecturer
長崎県 長崎県獣医師会産業動物臨床部会講習会 2023.10.18
Media Coverage 【 display / non-display 】
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よい方向へ正念場 Newspaper, magazine
日本農業新聞 日本農業新聞 現場からの農村学教室 2023.9.10
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家畜伝染病予防のための検査を可能にする環境に Promotional material
宮崎市 宮崎市広報 みらいの宮崎を創る人 フォーカス 2023.7.1
Author：Other
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宮大 格安検査準備進む Newspaper, magazine
宮崎日日新聞 2022.10.24
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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
Author：Myself
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獣医医事審議会専門員
Academic Activities 【 display / non-display 】
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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
Type：Academic research
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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
Type：Academic research