Mechanisms of the resistance against disease progression during bovine leukemia virus infection in cattle with DRB3*0902 allele

• Project/Area Number: 19H03128
• 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: University of Miyazaki
• Principal Investigator: Norimine Junzo 宮崎大学, 農学部, 教授 (30627667)
• Co-Investigator(Kenkyū-buntansha): 佐藤 克明 宮崎大学, 医学部, 教授 (40301147) ; 今内 覚 北海道大学, 獣医学研究院, 教授 (40396304)
• Project Period (FY): 2019-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥17,290,000 (Direct Cost: ¥13,300,000、Indirect Cost: ¥3,990,000); Fiscal Year 2022: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2021: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2020: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2019: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
• Keywords: 牛MHCクラスII / テトラマー / ヘルパーT細胞 / CD4+T細胞 / 牛白血病 / 牛伝染性リンパ腫 / プロウイルス量 / 病態進行 / 牛伝染性リンパ腫症 / bovine leukemia virus / MHCクラスII / DRB3*009:02 / 抵抗性 / アリル / CD4T細胞動態 / BLV抵抗性 / BoLAクラスIIテトラマー / CD4+T細胞エピトープ / DRB3*00902 / BoLA class II テトラマー / CD4T細胞 / BoLA クラスII テトラマー / エピトープ / 組換え蛋白 / CD4T細胞エピトープ / BLV構造蛋白 / BoLA / 発症抵抗性 / BLV / 抵抗性遺伝子 / MHC class II / 細胞性免疫

Outline of Research at the Start

日本では、BLV感染によって発症する牛白血病が毎年増加の一途をたどっている。また、本感染症に対する有効なワクチンも治療法も無いため、感染させない以外対策が無いのが現状である。BLVは、ヒトTリンパ向性ウイルス1型（HTLV-1）と近縁のレトロウイルスで、感染後の病態進行経過（癌化）においても両者は極めて類似している。BLV感染後の病態進行機序を解明する事により、他の類似するレトロウイルス感染における治療法の開発つながる可能性が期待できる。

Outline of Final Research Achievements

Enzootic bovine leukosis is an infectious disease caused by infection of bovine leukemia virus (BLV) belonging to Retroviridae family. Purpose of this study is to understand immunological mechanisms in which DRB3 genes have been thought to play a critical role for a protective immunity and are associated with reduction of proviral load and resistance against disease progression. Among DRB3 genes, the DRB3*009:02 allele is so far most strongly associated with the protection and its tetramer was therefore generated. The epitope mapping method using this tetramer have identified several BLV-specific T cell epitopes and made possible to analyze BLV-specific helper T cells. Although experimental infection using cattle have not been achieved by the time, the tetramer generated in this study have greatly enhanced the future opportunity to clarify the mechanisms.

Academic Significance and Societal Importance of the Research Achievements

牛伝染性リンパ腫ウイルス(BLV)による感染症は、ヒトT細胞白血病ウイルスI型(HTLV-1)による感染症と同様、その病態進行の機序は解明されていない。興味深い事に、BLVに感染してもDRB3*009:02アリルを保有する牛は、病態が進行しない。本研究では、このアリルの４量体（テトラマー）を作製し、これを利用してBLV特異的T細胞エピトープを同定した。牛DRB3*009:02のテトラマーが利用可能になった事で、このアリルが関与する抗原特異的防御免疫が解析できるようになった。特異的防御免疫の機序を明らかにすることは、類似する感染症の病態進行機序の解明そして治療法開発へつながることが期待できる。

Research Products

• [Journal Article] Identification of a DRB3*011:01-restricted CD4+ T cell response against bovine respiratory syncytial virus fusion protein (2023)
  • Author(s): Kaplan Bryan S.、Hofstetter Amelia R.、McGill Jodi L.、Lippolis John D.、Norimine Junzo、Dassanayake Rohana P.、Sacco Randy E.
  • Journal Title: Frontiers in Immunology Volume: 14 Pages: 1-14
  • DOI: 10.3389/fimmu.2023.1040075
  • Peer Reviewed
• [Journal Article] Development of droplet digital PCR for quantification of bovine leukemia virus proviral load using unpurified genomic DNA (2023)
  • Author(s): Wu Xinyue、Notsu Kousuke、Matsuura Yuichi、Mitoma Shuya、El Daous Hala、Norimine Junzo、Sekiguchi Satoshi
  • Journal Title: Journal of Virological Methods Volume: 315 Pages: 114706-114706
  • DOI: 10.1016/j.jviromet.2023.114706
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Identifying Pathogen and Allele Type Simultaneously in a Single Well Using Droplet Digital PCR (2023)
  • Author(s): Notsu Kosuke、El Daous Hala、Mitoma Shuya、Wu Xinyue、Norimine Junzo、Sekiguchi Satoshi
  • Journal Title: mSphere Volume: 8 Issue: 1 Pages: 1-13
  • DOI: 10.1128/msphere.00493-22
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A survey of bovine leukemia virus resistant bovine leukocyte antigen (<i>BoLA</i>)<i>-DRB3*009:02</i> allele-carrying Japanese Black cattle in two prefectures in Japan (2022)
  • Author(s): NOTSU Kosuke、INOUE Yoshinobu、MASUDA Tsuneyuki、INOUE Masahiro、MITOMA Shuya、WU Xinyue、NORIMINE Junzo、SEKIGUCHI Satoshi
  • Journal Title: Journal of Veterinary Medical Science Volume: 84 Issue: 11 Pages: 1457-1460
  • DOI: 10.1292/jvms.22-0285
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed / Open Access
• [Journal Article] A pooled testing system to rapidly identify cattle carrying the elite controller BoLA-DRB3*009:02 haplotype against bovine leukemia virus infection. (2022)
  • Author(s): Notsu K, El Daous H, Mitoma S, Norimine J, Sekiguchi S
  • Journal Title: HLA Volume: 99 Issue: 1 Pages: 12-24
  • DOI: 10.1111/tan.14502
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The detection of long-lasting memory foot-and-mouth disease (FMD) virus serotype O-specific CD4+ T cells from FMD-vaccinated cattle by bovine major histocompatibility complex class II tetramer. (2021)
  • Author(s): Mitoma S, Carr BV, Harvey Y, Moffat K, Sekiguchi S, Charleston B, Norimine J, Seago J
  • Journal Title: Immunology Volume: 164 Issue: 2 Pages: 266-278
  • DOI: 10.1111/imm.13367
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Relationship between Allelic Heterozygosity in BoLA-DRB3 and Proviral Loads in Bovine Leukemia Virus-Infected Cattle (2021)
  • Author(s): Daous Hala El、Mitoma Shuya、Elhanafy Eslam、Thi Nguyen Huyen、Thi Mai Ngan、Notsu Kosuke、Kaneko Chiho、Norimine Junzo、Sekiguchi Satoshi
  • Journal Title: Animals Volume: 11 Issue: 3 Pages: 647-647
  • DOI: 10.3390/ani11030647
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Characterization of bovine interleukin-2 stably expressed in HEK-293 cells (2021)
  • Author(s): MITOMA Shuya、EL-KHAIAT Heba M.、UTO Tomofumi、SATO Katsuaki、SEKIGUCHI Satoshi、NORIMINE Junzo
  • Journal Title: Journal of Veterinary Medical Science Volume: 83 Issue: 1 Pages: 134-141
  • DOI: 10.1292/jvms.20-0423
  • NAID: 130007971516
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Quantitative Risk Assessment for the Introduction of Bovine Leukemia Virus-Infected Cattle Using a Cattle Movement Network Analysis (2020)
  • Author(s): Kosuke Notsu, Anuwat Wiratsudakul, Shuya Mitoma, Hala El Daous, Chiho Kaneko, Heba M El-Khaiat, Junzo Norimine, Satoshi Sekiguchi
  • Journal Title: Pathogens Volume: 9(11) Issue: 11 Pages: 903-903
  • DOI: 10.3390/pathogens9110903
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Establishment of a novel diagnostic test for Bovine leukaemia virus infection using direct filter PCR (2020)
  • Author(s): El Daous Hala、Mitoma Shuya、Elhanafy Eslam、Thi Nguyen Huyen、Thi Mai Ngan、Hara Akihiro、Duangtathip Karn、Takezaki Yuka、Kaneko Chiho、Norimine Junzo、Sekiguchi Satoshi
  • Journal Title: Transboundary and Emerging Diseases Volume: 67 Issue: 4 Pages: 1671-1676
  • DOI: 10.1111/tbed.13506
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Direct filter PCR as a novel diagnostic test for bovine leukemia virus infection. (2019)
  • Author(s): El Daous, H., 三苫修也, Elhanafy E., NgyenThi H., Mai T. N., 原彰宏, Duangtathip K., 竹崎由佳, 兼子千穂, 乗峰潤三, 関口敏
  • Organizer: 第162回日本獣医学会学術集会
• [Presentation] 牛MHCクラスIIテテトラマをー用いた口蹄疫ウイルス構造蛋白エピトープマッピング (2019)
  • Author(s): 4.三苫修也、Julian Seago、Veronica Carr、Katy Maffot、Bryan Charleston、関口敏、乗峰潤三.
  • Organizer: 第162回日本獣医学会学術集会
• [Presentation] The role of BoLA-DRB3 alleles on disease progression during bovine leukemia virus infection. (2019)
  • Author(s): Norimine J
  • Organizer: HTLV 19th International conference on human retrovirology
  • Int'l Joint Research / Invited