Comprehensive analysis of Babesia tick-stage antigens: Establishing of a base for development the TBV

• Project/Area Number: 19H03120
• 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: KAWAZU Shin-ichiro 帯広畜産大学, 原虫病研究センター, 教授 (60312295)
• Co-Investigator(Kenkyū-buntansha): 白藤 梨可 帯広畜産大学, 原虫病研究センター, 准教授 (00549909) ; 麻田 正仁 帯広畜産大学, 原虫病研究センター, 准教授 (40587028) ; 山岸 潤也 北海道大学, 人獣共通感染症国際共同研究所, 准教授 (80535328)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000); Fiscal Year 2021: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2020: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2019: ¥6,760,000 (Direct Cost: ¥5,200,000、Indirect Cost: ¥1,560,000)
• Keywords: 獣医学 / 疾病予防・制御 / バベシア / マダニステージ / 遺伝子改変原虫

Outline of Research at the Start

本研究はバベシアのマダニ体内発育ステージに特異的な細胞表在性蛋白質を網羅的に検出し、その機能を解析するとともに伝搬阻止ワクチン（TBV）抗原としての性能を評価することを目的とする。バベシア病による経済損失は依然として深刻な問題で、現行の家畜体内（赤血球内）発育ステージを標的とするワクチンや創薬には限界がある。他方、バベシアのマダニ体内での発育ステージの研究は殆ど手つかずの状況にある。本研究では、網羅的手法を用いて同定したバベシアのマダニ体内発育ステージ表在性抗原について（１）原虫発育における機能を解析し（２）TBV抗原としての性能を評価する。

Outline of Final Research Achievements

Babesia ovata erythrocytic stage (Blood stage) was induced into the tick development stage (Tick stage) by an in vitro induction method, and RNA-sequence method was used to search for transmission-blocking vaccine (TBV) target molecules. The Tick stage at 6h after induction showed a characteristic form of the parasite called ray body, in which the expression of Tick stage genes was observed. The genes that were more abundantly expressed in the Tick stage than in the Blood stage were searched for genes that were coding for cell-surface proteins. As a result, 14 genes encoding proteins presumed to be cell surface-localization as potential TBV target molecules were identified.

Academic Significance and Societal Importance of the Research Achievements

節足動物を介して伝搬するマラリアでは、次世代型のワクチンとして、蚊体内での発育ステージを標的としたTBVが既に臨床試験の段階にある。他方、バベシアのマダニ体内での発育ステージの研究は、実験系の整備の遅れから殆ど手つかずの状況にある。本研究では、RNA-sequence法を用いてバベシアのマダニ体内発育ステージ表在性抗原を同定した。一連の研究成果から、バベシア病に対するTBVの開発研究が進展する。同時に、これら分子がマダニ体内での原虫発育に果たす役割を解明できれば、節足動物媒介性の原虫がどのように各々の宿主適応戦略を進化させてきたのか、という基本的な疑問にも分子論的な考察が加えられるようになる。

Research Products

• [Int'l Joint Research] ケレタロ自治大学(メキシコ)
• [Int'l Joint Research] チェコ科学アカデミー(CAS)(チェコ)
• [Int'l Joint Research] ケレタロ自治大学(メキシコ)
• [Int'l Joint Research] チェコ科学アカデミー(CAS)(チェコ)
• [Int'l Joint Research] ケレタロ自治大学(メキシコ)
• [Int'l Joint Research] チェコ科学アカデミー（CAS）(チェコ)
• [Journal Article] Advances in understanding red blood cell modifications by Babesia (2022)
  • Author(s): Hakimi H, Yamagishi J, Kawazu SI, Asada M
  • Journal Title: PLoS Pathogens Volume: 18(9) Issue: 9 Pages: e1010770-e1010770
  • DOI: 10.1371/journal.ppat.1010770
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The GP-45 Protein, a Highly Variable Antigen from Babesia bigemina, Contains Conserved B-Cell Epitopes in Geographically Distant Isolates (2022)
  • Author(s): Mercado-Uriostegui Miguel Angel、Castro-S?nchez Luis Alberto、Batiha Gaber El-Saber、Valdez-Espinoza Uriel Mauricio、Falc?n-Neri Alfonso、Ramos-Aragon Juan Alberto、Hern?ndez-Ortiz Ruben、Kawazu Shin-Ichiro、Igarashi Ikuo、Mosqueda Juan
  • Journal Title: Pathogens Volume: 11 Issue: 5 Pages: 591-591
  • DOI: 10.3390/pathogens11050591
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Recent Advances in Molecular Genetic Tools for Babesia (2021)
  • Author(s): Hassan Hakimi, H, Masahito Asada, Shinichiro Kawazu
  • Journal Title: Veterinary Sciences Volume: 8 Issue: 10 Pages: 222-222
  • DOI: 10.3390/vetsci8100222
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Isolation of viable Babesia bovis merozoites to study parasite invasion (2021)
  • Author(s): Hassan Hakimi, H, Masahito Asada, Takahiro Ishizaki, Shinichiro Kawazu
  • Journal Title: Scientific reports Volume: 11 Issue: 1 Pages: 16959-16959
  • DOI: 10.1038/s41598-021-96365-w
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Evaluation of 5-aminolevulenic acid plus ferrous ion for its potential to improve symptoms of bovine babesiosis. (2021)
  • Author(s): Elata, A., Suganuma, K., Mossaad, E., Zafar, I., Galon, EM., Yoshimoto, K., Tanaka, T., Xuenan, X., and Kawazu, SI.
  • Journal Title: Parasitologia Volume: 1(4) Issue: 4 Pages: 218-224
  • DOI: 10.3390/parasitologia1040023
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Genome editing of Babesia bovis using the CRISPR/Cas9 system. (2019)
  • Author(s): Hakimi, H., Ishizaki, T., Kegawa, Y., Kaneko, O., Kawazu, S., and Asada, M.
  • Journal Title: mSphere Volume: 4(3) Issue: 3
  • DOI: 10.1128/msphere.00109-19
  • NAID: 120006988459
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Initiated Babesia ovata Sexual Stages under In Vitro Conditions Were Recognized by Anti-CCp2 Antibodies, Showing Changes in the DNA Content by Imaging Flow Cytometry (2019)
  • Author(s): Nguyen TT, Dang-Trinh MA, Higuchi L, Mosqueda J, Hakimi H, Asada M, Yamagishi J, Umemiya-Shirafuji R, Kawazu SI
  • Journal Title: Pathogens Volume: 8(3) Issue: 3 Pages: 104-104
  • DOI: 10.3390/pathogens8030104
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] RON2, a novel gene in Babesia bigemina, contains conserved, immunodominant B-cell epitopes that induce antibodies that block merozoite invasion. (2019)
  • Author(s): Mosqueda, J., Hidalgo-Ruiz, M., Calvo-Olvera, DA., Hernandez-Silva, DJ., Ueti, MW., Mercado-Uriostegui, MA., Rodriguez, A., Ramos-Aragon, JA., Hernandez-Ortiz, R., Kawazu, SI., and garashi, I.
  • Journal Title: Parasitology Volume: 146(13 Issue: 13 Pages: 1646-1654
  • DOI: 10.1017/s0031182019001161
  • Peer Reviewed / Int'l Joint Research
• [Presentation] A novel Babesia bovis secreted protein responsible for binding of infected erythrocyte to endothelial cell. (2021)
  • Author(s): 晴希生ハッサン、坂口美亜子、山岸潤也、河津信一郎、金子修、麻田正仁
  • Organizer: 第90回日本寄生虫学会大会
• [Presentation] Babesia bovis TRAP関連分子p200は赤血球期増殖に重要な役割を担う (2021)
  • Author(s): 麻田正仁、晴希生ハッサン、金子修、河津信一郎
  • Organizer: 第90回日本寄生虫学会大会
• [Presentation] Cyclic GMP-dependent protein kinase plays a central role for egress of Babesia bovis. (2021)
  • Author(s): Hassan H., Asada M., and Kawazu S.
  • Organizer: 第164回日本獣医学会
• [Presentation] TRAP関連分子p200は赤血球期増殖に重要な役割を担う (2021)
  • Author(s): 麻田正仁、晴希生ハッサン、金子修、河津信一郎
  • Organizer: 第164回日本獣医学会
• [Presentation] Babesia ovata tick stages induced under in vitro conditions: Observation of morphological and ploidy changes. (2019)
  • Author(s): Thu-Thuy Nguyen、Minh Anh Dang Trinh、樋口留菜、Juan Mosqueda、Hassan Hakimi、麻田正仁、山岸潤也、梅宮-白藤梨可、河津信一郎
  • Organizer: 第162回日本獣医学会
• [Presentation] The expression of a novel multigene family is correlated with channel activity in Babesia bovis-infected erythrocytes. (2019)
  • Author(s): Hakimi H., Templeton TJ., Sakaguchi M., Yamagishi J., Yahata K., Kawazu S., Kaneko O., and Asada M.
  • Organizer: 第89回日本寄生虫学会大会 口頭発表（集会中止・誌上発表）
• [Presentation] 放牧地および自然環境の境界域におけるマダニ類の局所的な種多様性の比較 (2019)
  • Author(s): 尾針由真、白藤（梅宮）梨可、河津信一郎
  • Organizer: 89回日本寄生虫学会大会 ポスター（集会中止・誌上発表）
• [Remarks] 帯広畜産大学 原虫病研究センター （日本語版）
  • URL: https://www.obihiro.ac.jp/facility/protozoa/
• [Remarks] 帯広畜産大学 原虫病研究センター （英語版）
  • URL: https://www.obihiro.ac.jp/facility/protozoa/en