家畜病原性原虫ネオスポラの感染による流産発症機構の解明

• Project/Area Number: 18H02335
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 42020:Veterinary medical science-related
• Research Institution: Obihiro University of Agriculture and Veterinary Medicine
• Principal Investigator: 西川 義文 帯広畜産大学, 原虫病研究センター, 教授 (90431395)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000); Fiscal Year 2020: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2019: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000); Fiscal Year 2018: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
• Keywords: 原虫 / ネオスポラ / 流産 / 垂直感染 / 病原性

Outline of Annual Research Achievements

【流産誘発因子破壊原虫を用いたマウス感染実験による妊娠期の病態解析】原虫因子破壊株（NcGRA7KO株）を非妊娠雌マウスへ感染させ、マウスの生存率、原虫の体内分布、組織を調べたところ、親株（Nc1株）と比較してNcGRA7KO株の病原性が低下していた。次に妊娠期感染マウスモデルにてNcGRA7KO株の病原性を解析し、垂直感染の低下が認められた。NcGRA7KO株感染マウスの胎盤組織では炎症反応の亢進が認められた。従って、胎盤で誘導された抗原虫反応により、NcGRA7KO株の妊娠期の病原性が低下したと推測された。
【流産誘発因子と相互作用する宿主因子の同定】 FLAGタグ融合原虫因子（NcGRA7-FLAG）を導入した哺乳動物細胞（293T細胞）とFLAGタグ融合原虫因子を遺伝子欠損原虫へ導入した原虫株（NcGRA7Comp株）を用いて、質量分析によりNcGRA7結合タンパク質の候補タンパク質Xを同定した。タンパク質XをsiRNA法により免疫細胞からノックダウンし、ネオスポラ感染による免疫応答を解析したところ、炎症反応の減少が認められた。従って、感染による免疫反応におけるタンパク質Xの重要性が確認された。
【ネオスポラ流産牛の組織サンプルを用いた流産誘発因子の同定】ネオスポラ症のウシ由来組織サンプル（胎盤、流産胎児、神経症状等）におけるNcGRA7の発現・局在を解析した。原虫が増殖した寄生胞にてNcGRA7の発現が確認され、壊死像にもNcGRA7のシグナルが認められた。さらに、感染した神経細胞にてNcGRA7が寄生胞から細胞質へ分泌される特徴的な局在が確認された。NcGRA7を診断用抗原としたELISA系はすでに構築しており、ネオスポラ流産牛の血清ではNcGRA7抗体レベルが上昇していた。このことから、NcGAR7がネオスポラ流産に関与することが推測された。

Research Progress Status

令和2年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

令和2年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] South Valley University/Mansoura University(エジプト)
• [Int'l Joint Research] モンゴル生命科学大学(モンゴル)
• [Int'l Joint Research] マヒドン大学(タイ)
• [Int'l Joint Research] South Valley University/Mansoura University(エジプト)
• [Int'l Joint Research] University of the Philippines Cebu/Cebu Technological University/University of the Visayas(フィリピン)
• [Int'l Joint Research] South Valley大学(エジプト)
• [Int'l Joint Research] モンゴル生命科学大学(モンゴル)
• [Int'l Joint Research] マヒドン大学(タイ)
• [Journal Article] Role of dense granule antigen 7 in vertical transmission of Neospora caninum in C57BL/6 mice infected during early pregnancy (2022)
  • Author(s): Abdou AM, Ikeda R, Watanabe K, Furuoka H, Nishikawa Y
  • Journal Title: Parasitol Int Volume: 89 Pages: 102576-102576
  • DOI: 10.1016/j.parint.2022.102576
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Comparative Evaluation of Four Potent Neospora caninumDiagnostic Antigens Using Immunochromatographic Assay for Detection of Specific Antibody in Cattle (2021)
  • Author(s): Fereig RM, Abdelbaky HH, Nishikawa Y
  • Journal Title: Microorganisms Volume: 9 Pages: 2133-2133
  • DOI: 10.3390/microorganisms9102133
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Genetic characterization of Neospora caninum from aborted bovine fetuses in Hokkaido, Japan (2021)
  • Author(s): El-Alfy ES, Ohari Y, Shimoda N, Nishikawa Y
  • Journal Title: Infect Genet Evol Volume: 92 Pages: 104838-104838
  • DOI: 10.1016/j.meegid.2021.104838
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Vaccination with Neospora GRA6 Interrupts the Vertical Transmission and Partially Protects Dams and Offspring against Neospora caninum Infection in Mice. (2021)
  • Author(s): Fereig RM, Abdelbaky HH, Nishikawa Y.
  • Journal Title: Vaccines Volume: 9 Pages: 155-155
  • DOI: 10.3390/vaccines9020155
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Urological detection of specific antibodies against Neospora caninum infection in mice: A prospect for novel diagnostic approach of Neospora. (2020)
  • Author(s): Fereig RM, Nishikawa Y.
  • Journal Title: Exp Parasitol Volume: 216 Pages: 107942-107942
  • DOI: 10.1016/j.exppara.2020.107942
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Evaluation of Neospora caninum serodiagnostic antigens for bovine neosporosis. (2020)
  • Author(s): Abdelbaky HH, Nishimura M, Shimoda N, Hiasa J, Fereig RM, Tokimitsu H, Inokuma H, Nishikawa Y.
  • Journal Title: Parasitol Int Volume: 75 Pages: 102045-102045
  • DOI: 10.1016/j.parint.2019.102045
  • NAID: 120007004078
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] From Signaling Pathways to Distinct Immune Responses: Key Factors for Establishing or Combating Neospora caninum Infection in Different Susceptible Hosts. (Review) (2020)
  • Author(s): Fereig RM, Nishikawa Y.
  • Journal Title: Pathogens. Volume: 9 Pages: 384-384
  • DOI: 10.3390/pathogens9050384
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Seroepidemiological study of Toxoplama gondii in small ruminants (sheep and goat) in different provinces of Mongolia. (2020)
  • Author(s): Pagmadulam B, Myagmarsuren P, Yokoyama N, Battsetseg B, Nishikawa Y.
  • Journal Title: Parasitol Int. Volume: 74 Pages: 101996-101996
  • DOI: 10.1016/j.parint.2019.101996
  • NAID: 120006955802
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Metacytofilin is a potent therapeutic drug candidate for toxoplasmosis. (2020)
  • Author(s): Leesombun A, Iijima M, Umeda K, Kondoh D, Pagmadulama B, Abdou AM, Suzuki Y, Ohba S, Isshiki K, Kimura T, Kubota Y, Sawa R, Nihei C, Nishikawa Y.
  • Journal Title: J Infect Dis. Volume: 221 Pages: 766-774
  • DOI: 10.1093/infdis/jiz501
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Transcriptome analysis of the effect of C-C chemokine receptor 5 deficiency on cell response to Toxoplasma gondii in brain cells (2019)
  • Author(s): Kobayashi K, Umeda K, Ihara F, Tanaka S, Yamagishi J, Suzuki Y, Nishikawa Y.
  • Journal Title: BMC Genomics. Volume: 20 Pages: 705-705
  • DOI: 10.1186/s12864-019-6076-4
  • NAID: 120006732636
  • Peer Reviewed / Open Access
• [Journal Article] Specific increase of Fusobacterium in the faecal microbiota of neonatal calves infected with Cryptosporidium parvum (2019)
  • Author(s): Ichikawa-Seki Madoka、Motooka Daisuke、Kinami Aiko、Murakoshi Fumi、Takahashi Yoko、Aita Junya、Hayashi Kei、Tashibu Atsushi、Nakamura Shota、Iida Tetsuya、Horii Toshihiro、Nishikawa Yoshifumi
  • Journal Title: Sci Rep. Volume: 9 Pages: 12517-12517
  • DOI: 10.1038/s41598-019-48969-6
  • NAID: 120007175164
  • Peer Reviewed / Open Access
• [Journal Article] Isolation and characterization of antiprotozoal compound-producing Streptomyces species from Mongolian soils. (2019)
  • Author(s): Pagmadulam B, Tserendulam D, Rentsenkhand T, Igarashi M, Sawa R, Nihei CI, Nishikawa Y.
  • Journal Title: Parasitol Int. Volume: 74 Pages: 101961-101961
  • DOI: 10.1016/j.parint.2019.101961
  • NAID: 120006955803
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Involvement of Toll-like receptor 2 in the cerebral immune response and behavioral changes caused by latent Toxoplasma infection in mice (2019)
  • Author(s): Fumiaki Ihara, Sachi Tanaka, Ragab M Fereig, Maki Nishimura, Yoshifumi Nishikawa
  • Journal Title: PLoS One. Volume: 14
  • DOI: 10.1371/journal.pone.0220560
  • NAID: 120006732638
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Molecular screening approach to identify protozoan and trichostrongylid parasites infecting one-humped camels (Camelus dromedarius). (2019)
  • Author(s): El-Alfy ES, Abu-Elwafa S, Abbas I, Al-Araby M, Al-Kappany Y, Umeda K, Nishikawa Y.
  • Journal Title: Acta Trop Volume: 197 Pages: 105060-105060
  • DOI: 10.1016/j.actatropica.2019.105060
  • NAID: 120006713048
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Endemicity of Toxoplasma infection and its associated risk factors in Cebu, Philippines. (2019)
  • Author(s): Ybanez RHD, Busmeon CGR, Viernes ARG, Langbid JZ, Nuevarez JP, Ybanez AP, Nishikawa Y
  • Journal Title: PLoS One. Volume: 14
  • DOI: 10.1371/journal.pone.0217989
  • NAID: 120006713049
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Ethanol Extracts from Thai Plants have Anti-Plasmodium and Anti-Toxoplasma Activities In Vitro. (2019)
  • Author(s): Leesombun A, Boonmasawai S, Nishikawa Y.
  • Journal Title: Acta Parasitol. Volume: 64 Pages: 257-261
  • DOI: 10.2478/s11686-019-00036-w
  • NAID: 120006727120
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Neospora GRA6 possesses immune-stimulating activity and confers efficient protection against Neospora caninum infection in mice. (2019)
  • Author(s): Fereig RM, Shimoda N, Abdelbaky HH, Kuroda Y, Nishikawa Y.
  • Journal Title: Vet Parasitol. Volume: 267 Pages: 61-68
  • DOI: 10.1016/j.vetpar.2019.02.003
  • NAID: 120006603078
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Critical role of TLR2 in triggering protective immunity with cyclophilin entrapped in oligomannose-coated liposomes against Neospora caninum infection in mice. (2019)
  • Author(s): Fereig RM, Abdelbaky HH, Kuroda Y, Nishikawa Y.
  • Journal Title: Vaccine Volume: 37 Pages: 937-944
  • DOI: 10.1016/j.vaccine.2019.01.005
  • NAID: 120006583709
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Seroprevalence of Cryptosporidium parvum and Neospora caninum in cattle in the southern Kyushu region of Japan. (2018)
  • Author(s): Masatani T, Fereig RM, Otomaru K, Ishikawa S, Kojima I, Hobo S, Nishikawa Y.
  • Journal Title: Parasitol Int. Volume: 67 Pages: 763-767
  • DOI: 10.1016/j.parint.2018.08.001
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Seroprevalence of Toxoplasma gondii and Neospora caninum infections in cattle in Mongolia. (2018)
  • Author(s): Pagmadulam B, Myagmarsuren P, Fereig RM, Igarashi M, Yokoyama N, Battsetseg B, Nishikawa Y.
  • Journal Title: Vet Parasitol Reg Stud Reports. Volume: 14 Pages: 11-17
  • DOI: 10.1016/j.vprsr.2018.08.001
  • NAID: 120006533723
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Neospora caninum dense granule protein 7 regulates pathogenesis of neosporosis by modulating host immune response. (2018)
  • Author(s): Nishikawa Y*, Shimoda N, Fereig RM, Moritaka T, Umeda K, Nishimura M, Ihara F, Kobayashi K, Himori Y, Suzuki Y, Furuoka H.
  • Journal Title: Appl Environ Microbiol. Volume: 84
  • DOI: 10.1128/aem.01350-18
  • NAID: 120006533724
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Identification of the antigenic region of Neospora caninum dense granule protein 7 using ELISA. (2018)
  • Author(s): Abdelbaky HH, Fereig RM, Nishikawa Y
  • Journal Title: Parasitol Int. Volume: 67 Pages: 675-678
  • DOI: 10.1016/j.parint.2018.06.012
  • NAID: 120006769770
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Current situation of Neospora caninum and Toxoplasma gondii infection among beef cattle in Kanchanaburi, Ratchaburi and Nakhon Patom provinces, Thailand. (2018)
  • Author(s): Ruenruetai Udonsom, Yaowalark Sukthana, Yoshifumi Nishikawa, Ragab M. Fereig, Charoonluk Jirapattharasate.
  • Journal Title: The Thai Journal of Veterinary Medicine Volume: 48 Pages: 403-409
  • Peer Reviewed / Int'l Joint Research
• [Presentation] Role of dense granule antigen 7 on vertical transmission of Neospora caninum in mice (2021)
  • Author(s): Abdou Ahmed、池田 里奈、渡邉 謙一、古岡 秀文、西川 義文
  • Organizer: 第90回日本寄生虫学会大会
• [Presentation] Genetic characterization of Neospora caninum from aborted bovine fetuses in Hokkaido, Japan (2021)
  • Author(s): El-Sayed El-Alfy, Yuma Ohari, Naomi Shimoda, Yoshifumi Nishikawa
  • Organizer: British Society for Parasitology, Parasites Online 2021
  • Int'l Joint Research
• [Presentation] Immunization with Neospora GRA6 confers efficient protection against Neospora caninum infection in both non-pregnant and pregnant mice (2021)
  • Author(s): Ragab Fereig、下田 直美、Abdelbaky Hanan、西川 義文
  • Organizer: 第164回日本獣医学会学術集会
• [Presentation] Possible effects of dense granule protein 7 on vertical transmission of Neospora caninum (2020)
  • Author(s): Abdou Ahmed、池田里奈、古岡秀文、西川義文
  • Organizer: 第８９回日本寄生虫学会大会
• [Presentation] Identification of Neospora caninum serodiagnostic antigens for bovine neosporosis (2020)
  • Author(s): Abdelbaky Hanan、西村麻紀、下田直美、日浅淳、Fereig Ragab、時光宏三、猪熊壽、西川義文
  • Organizer: 第８９回日本寄生虫学会大会
• [Presentation] 妊娠期トキソプラズマ感染の病態発症機序の解明 (2019)
  • Author(s): 西川義文
  • Organizer: 第２7回分子寄生虫学ワークショップ／第１7回分子寄生虫・マラリア研究フォーラム合同大会
• [Presentation] Toxoplasma gondii conducts NFκB pathway of host cells, dense granule protein 14 is a novel member of the orchestra (2019)
  • Author(s): Fumiaki Ihara, Kosuke Umeda, Masahiro Yamamoto, Yoshifumi Nishikawa.
  • Organizer: 第18回あわじ感染と免疫国際フォーラム
  • Int'l Joint Research
• [Presentation] Neospora caninum dense granule protein 7 regulates pathogenesis of neosporosis by modulating host immune response. (2019)
  • Author(s): 西川義文、下田直美、Ragab Fereig、森高智弥、梅田剛佑、西村麻紀、猪原史成、小林薫、檜森結羽、鈴木穣、古岡秀文
  • Organizer: 第162回日本獣医学会学術集会
• [Presentation] Effect of metacytofilin against Toxoplasma gondii. (2019)
  • Author(s): Arpron Leesombun、飯島正富、梅田剛佑、近藤大輔、Baldorj Pagmadulam、Ahmed Abdou、鈴木穣、大庭俊一、一色邦夫、木村智之、久保田由美子、澤竜一、二瓶浩一、西川義文
  • Organizer: 第162回日本獣医学会学術集会
• [Presentation] Development of potent and safe vaccine candidates against Neospora caninum infection. (2018)
  • Author(s): Ragab Fereig、西川義文
  • Organizer: 第162回日本獣医学会学術集会
  • Invited
• [Presentation] Sero-epidemiological study of Toxoplasma gondii and Neospora caninum infections in cattle in Mongolia. (2018)
  • Author(s): Pagmadulam Baldorj、Myagmarsuren Punsantsogvoo、Ragab Fereig、五十嵐 慎、横山 直明、Battsetseg Badgar、西川 義文
  • Organizer: 第161回日本獣医学会学術集会
• [Presentation] Neospora caninumの病原性因子の同定 (2018)
  • Author(s): 西川義文
  • Organizer: 第２6回分子寄生虫学ワークショップ／第１6回分子寄生虫・マラリア研究フォーラム合同大会
• [Presentation] Evaluation of serodiagnostic antigens of Neospora caninum for bovine neosporosis. (2018)
  • Author(s): Hanan H. Abdelbaky, Maki Nishimura, Naomi Shimoda, Jun Hiasa, Hiromi Tokimitsu, Hisashi Inokuma, Yoshifumi Nishikawa
  • Organizer: 第 64 回日本寄生虫学会・日本衛生動物学会 北日本支部合同大会
• [Presentation] ネオスポラ症の病態に関与するNcGRA6 の機能解析 (2018)
  • Author(s): 森高智弥, 下田直美, 西川義文
  • Organizer: 第 64 回日本寄生虫学会・日本衛生動物学会 北日本支部合同大会
• [Book] Methods Mol Biol (2022)
  • Author(s): Fereig RM, Nishikawa Y
  • Total Pages: 360
  • Publisher: Humana New York, NY
  • ISBN: 9781071618882
• [Book] 寄生虫病学 第３版 (2020)
  • Author(s): 日本獣医寄生虫学会
  • Total Pages: 276
  • Publisher: 緑書房
  • ISBN: 9784895315692
• [Remarks] 帯広畜産大学・原虫病研究センター・西川研究室
  • URL: https://sites.google.com/site/nishihdlab/
• [Remarks] 帯広畜産大学・原虫病研究センター
  • URL: http://www.obihiro.ac.jp/~protozoa/index.html
• [Remarks] 帯広畜産大学・原虫病研究センター・生体防御学分野・西川研究室
  • URL: https://sites.google.com/site/nishihdlab/
• [Remarks] 帯広畜産大学
  • URL: https://www.obihiro.ac.jp
• [Remarks] 未踏の領野に挑む、知の開拓者たち vol.61
• [Remarks] http://shochou-kaigi.org/interview/interview_61/