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Development of model system to analyze the antigen epitope to induce cell-mediated immunity in fish

Research Project

Project/Area Number 12460088
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field General fisheries
Research InstitutionNihon University

Principal Investigator

NAKANISHI Teruyuki  Biological Sciences, veterinary Medicine, Professor, 生物資源科学部, 教授 (00322496)

Co-Investigator(Kenkyū-buntansha) MORITOMO Tadaaki  Biological Sciences, veterinary Medicine, Associate Professor, 生物資源科学部, 助教授 (20239677)
OKAMOTO Nobuaki  Tokyo University of Fisheries, Professor, 水産学部, 教授 (40114912)
Project Period (FY) 2000 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥12,900,000 (Direct Cost: ¥12,900,000)
Fiscal Year 2002: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2001: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2000: ¥6,900,000 (Direct Cost: ¥6,900,000)
KeywordsCell-mediated immunity / Cytotoxicity test / MHC class I / IHN virus / DNA vaccination / Rainbow trout / Ginbuna crucian carp / Mixed leukocyte culture / )ニジマス / DNAフィンープリント法 / モノクローナル抗体 / トランスフェクション / MHC クラスI / ウイルス / 長期培養細胞 / ギンブナ / ヒラメ
Research Abstract

Fish possess specific antigen recognition molecules as well as B and T lymphocytes and can evoke specific immune responses against a variety of antigens. In fish, however, information on cell-mediated immunity is limited when compared to those on humoral immunity due to the lack of in vivo and in vitro model systems. Vaccine against viral disease is limited in fish despite that viral diseases are serious problem in aquaculture. Development of such vaccines requires identification and characterization of viral antigens which induce protective immunity. It is well known that cell-mediated killing is most important defense mechanisms in the control of viral disease. The objective of this research project is to develop experimental model systems to analyze the protective antigens or epitopes involved in cell-mediated immunity for the development of fish vaccines.
In collaboration with Dr. Okamoto at Tokyo University of Fisheries and Dr. Ototake at National Research Institute of Aquaculture … More we have established several assay systems for specific cell-mediated immunity, particularly on cytotoxic T-cell function employing clonal ginbuna and a syngeneic cell line. Those include the graft-versus host reaction (GVHR) and cell-mediated cytotoxicity (CMC) against allogeneic erythrocytes or cell lines. In the present research project we further pursued the studies and have established model systems for CMC against virus-infected cells and analyzed the protection mechanism using isogeneic ginbuna and cell lines together with fish viruses. We also extensively analyzed the MHC polymorphism in rainbow trout and found that trout cell line (RTG-2) shares the same MHC class I allele with homozygous clonal rainbow trout (C25 clone). We demonstrated MHC-restricted cell-mediated cytotoxicity against virus-infected cell lines using lymphocytes from C25 clonal trout sensitized by DNA immunization with IHNV-G or by infection with IHNV and RTG-2 cells as target. Experimental model systems established in the present study can be applied to investigate the protective antigens or epitopes involved in cell-mediated immunity for the development of fish vaccines as well as to investigate the recognition and killing g mechanism of specific cell-mediated immunity. Less

Report

(4 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • 2000 Annual Research Report
  • Research Products

    (43 results)

All Other

All Publications (43 results)

  • [Publications] Xia, C., Kiryu, I., J.M.Dijkstra, T.Azuma, T.Nakanishi, M.Ototake: "Differences in MHC class I between strains of rainbow trout (Oncorhynchus mykiss)"Fish & Shellfish Immunol.. 12. 287-301 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Okubo, K., Qin Qiwei, K.Aoyagi, K.Yamano, M.Ototake, T.Nakanishi, H.Fukuda, J.M.Dijkstra: "Ubiquitin genes in rainbow trout (Oncorhynchus mykiss)"Fish & Shellfish Immunol.. 12. 335-351 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Somamoto, T., T.Nakanishi, N Okamoto: "Roles of specific cell-mediated cytotoxicity in protecting fish from viral infections"Virology. 297. 120-127 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 乙竹充, J.M.Dijkstra, 桐生郁也, 吉浦康寿, 藤原篤志, U.Fischer, 中西照幸: "ニジマスMHC遺伝子の多様性及び機能の解析"水産育種. 32. 67-74 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Dijkstra, J.M., B.Kollner, K.Aoyagi, I.Denda, A.Kuroda, M.Ototake, K.Hashimoto, T.Nakanishi, Fischer, U.: "The rainbow trout classical MHC class I molecule Onmy-UBA ^*501 is expressed in similar cell types as mammalian classical MHC class I molecules"Fish & Shellfish Immunol.. 14. 1-23 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] J.M.Dijkstra, Y.Yoshiura, I.Kiryu, K.Aoyagi, B.Kollner, U.Fischer, T.Nakanishi, M.Ototake: "The promoter of the classical MHC class I locus in rainbow trout (Oncorhynchus mykiss)"Fish & Shellfish Immunol.. 14. 177-185 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nakanishi, T., U.Fischer, J.M.Dijkstra, M.Ototake: "MHC-restricted cell-mediated cytotoxicity in fish"Fisheries Science Vol.68, Suppl.. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Somamoto, T., T.Nakanishi, N.Okamoto: "Specific cytotoxic cells of ginbuna crucian carp are responsible for in vivo clearance of carp haematopoietic necrosis virus(CHNV)"Fisheries Science Vol.68, Suppl.. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 中西照幸, J.M.Dijkstra, 桐生郁也, 乙竹充: "魚類の免疫系(免疫系の遺伝子、8.主要組織適合遺伝子複合体(MHC)"恒星社厚生閣. 163(114-127) (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Somamoto, T., Nakanishi, T. and Okamoto, N.: "Specific cell-mediated cytotoxicity against a virus-infected syngeneic cell line in isogeneic ginbuna crucian carp"Dev. Comp. Immunol.. 24. 633-640 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Dijkstra, J.M., U. Fischer, I. Denda, M. Ototake and T. Nakanishi: "Exogenous antigens and the stimulation of MHC class I restricted cell-mediated cytotoxicity: possible strategies for fish vaccines"Fish & Shellfish Immunol.. 11. 437-458 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nakanishi, T., U. Fischer, J.M. Dijkstra, S. Hasegawa, T. Somamoto, N. Okamoto and M. Ototake: "Cytotoxic T cell function in fish"Dev. Comp. Immunol.. 26. 131-139 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Aoyagi K., J.M. Dijkstra, C. Xia, I. Denda, M. Ototake, K Hashimoto, and T. Nakanishi: "Classical MHC class I genes composed of highly divergent sequence lineages share a single locus in rainbow trout (Oncorhynchus mykiss)"J. Immunology. 168. 260-273 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Xia, C., Kiryu, I., J.M. Dijkstra, T. Azuma, T. Nakanishi and M. Ototake: "Differences in MHC class I between strains of rainbow trout (Oncorhynchus mykiss)"Fish & Shellfish Immunol.. 12. 287-301 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Okubo, K., Qin Qiwei, K. Aoyagi, K. Yamano, M. Ototake, T. Nakanishi, H. Fukuda and J. M. Dijkstra: "Ubiquitin genes in rainbow trout (Oncorhynchus mykiss)"Fish & Shellfish Immunol.. 12. 335-351 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Q. W. Qin, M. Ototake, H. Nagoya and T. Nakanishi: "Graft-versus-host reaction (GVHR) in clonal amago salmon, Oncorhunchus rhodurus"Vet. Immunol. Immunopathol.. 89. 83-89 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Somamoto, T., T. Nakanishi, and N Okamoto: "Roles of specific cell-mediated cytotoxicity in protecting fish from viral infections"Virology. 297. 120-127 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Dijkstra, J.M., B. Kollner, K. Aoyagi, I. Denda, A. Kuroda, M. Ototake, K. Hashimoto: "The rainbow trout classical MHC class I molecule Onmy-UBA*501 is expressed in similar cell types as mammalian classical MHC class I molecules"Fish & Shellfish Immunol.. 14. 1-23 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Ototake, M., J.M. Dijkstra, I. Kiryu, Y. Yoshiura, A. Fujiwara, U. Fischer and T. Nakanishi: "Structure and functions of MHC class I genes in rainbow trout Oncorhynchus mykiss"Fish Genetics and Breeding Science. 32. 67-74 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Dijkstra, J. M., Y. Yoshiura, I. Kiryu, K. Aoyagi, B. Kollner, U. Fischer, T. Nakanishi and M. Ototake: "The promoter of the classical MHC class I locus in rainbow trout (Oncorhynchus mykiss)"Fish & Shellfish Immunol.. 14. 177-185 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nakanishi, T., U. Fischer, J.M. Dijkstra and M. Ototake: "MCH-restricted cell-mediated cytotoxicity in fish"Fisheries Science. Vol. 68, Supple. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Somamoto, T., T. Nakanishi and N. Okamoto: "Specific cytotoxic cells of ginbuna crucian carp are responsible for in vivo clearance of carp haematopoietic necrosis virus (CHNV)"Fisheries Science. Vol. 68, Supple. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Xia, C., Kiryu, I., J.M.Dijkstra, T.Azuma, T.Nakanishi, M.Ototake: "Differences in MHC class I between strains of rainbow trout (Oncorhynchus mykiss)"Fish & Shellfish Immunol.. 12. 287-301 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Okubo, K., Qin Qiwei, K.Aoyagi, K.Yamano, M.Ototake, T.Nakanishi, H.Fukuda, J.M.Dijkstra: "Ubiquitin genes in rainbow trout (Oncorhynchus mykiss)."Fish & Shellfish Immunol.. 12. 335-351 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Somamoto, T., T.Nakanishi, N Okamoto: "An in vitro and in vivo roles of specific cell-mediated cytotoxicity in protecting fish from viral infections"Virology. 297. 120-127 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Dijkstra, J.M., B.Kollner, K.Aoyagi, I.Denda, A.Kuroda, M.Ototake, K.Hashimoto, T.Nakanishi, Fischer, U.: "The rainbow trout classical MHC class I molecule Onmy-UBA^*501 is expressed in similar cell types as mammalian classical MHC class I molecules"Fish & Shellfish Immunol.. 14. 1-23 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] 乙竹充, J.M.Dijkstra, 桐生郁也, 吉浦康寿, 藤原篤志, U.Fischer, 中西照幸: "ニジマスMHC遺伝子の多様性及び機能の解析"水産育種. 32. 67-74 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Nakanishi, T., U.Fischer, J.M Dijkstra, M.Ototake: "MHC-restricted cell-mediated cytotoxicity in fish."Fisheries Science. 168(印刷中). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] J.M.Dijkstra, Y.Yoshiura, I.Kiryu, K.Aoyagi, B.Kollner, U.Fischer, T.Nakanishi, M.Ototake: "The promoter of the classical MHC class I locus in rainbow trout (Oncorhynchus mykiss)"Fish & Shellfish Immunol.. (印刷中). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] 中西照幸, J.M.Dijkstra, 桐生郁也, 乙竹充: "魚類の免疫系(IV.免疫系の遺伝子、8.主要組織適合遺伝子複合体(MHC))"恒星社厚生閣. 163 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Dijkstra, J.M., U.Fischer, I.Denda, M.Ototake, T.Nakanishi: "Exogenous antigens and the stimulation of MHC class I restricted cell-mediated cytotoxicity:possible strategies for fish vaccines"Fish & Shellfish Immunol.. 11. 437-458 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Nakanishi, T., U.Fischer, J.M.Dijkstra, S.Hasegawa, T.Somamoto, N.Okamoto, M.Ototake: "Cytotoxic T cell function in fish"Dev.Comp.Immunol.. 26. 131-139 (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] J.Immunology. 168. 260-273 (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Fish & Shellfish Immunol.. 12(印刷中). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Fish & Shellfish Immunol.. 12(印刷中). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Dijkstra, J.M., B.Kollner, K.Aoyagi, I.Denda, A.Kuroda, M.Ototake, K.Hashimoto, T.Nakanishi, Fischer, U.: "The rainbow trout classical MHC class I molecule Onmy-UBA^*501 is expressed in similar cell types as mammalian classical MHC class I molecules"Fish & Shellfish Immunol.. 12(印刷中). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Somamoto,T.,Nakanishi,T.and Okamoto,N.: "Specific cell-mediated cytotoxicity against a virus-infected syngeneic cell line in isogeneic ginbuna crucian carp."Dev.Comp.Immunol.. 24. 633-640 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Dijkstra,J.M.,H.Okamoto,M.Ototake,and T.Nakanishi: "Luciferase expression 2 years after DNA injection in glass catfish (Kryptopterus bicirrhus)."Fish & Shellfish Immunol.. 11. 199-202 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Somamoto T,Nakanishi T,Okamoto N.: "Cell-mediated cytotoxicity of ginbuna crucian carp after infection with crucian carp haematopoietic necrosis virus (CHNV)."Dev.Comp.Immunol.. 24. 7 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Dijkstra JM,Kollner B,Fischer U,Aoyagi K,Kuroda A,Ototake M,Nakanishi T.: "The rainbow trout classical MHC class I molecule Onmy-UBA^*501 is N-glycosylated and expressed in similar cell types as mammalian classical MHC class I molecules."Dev.Comp.Immunol.. 24. 75 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kollner B,Dijkstra JM,Fischer U,Schuetze H,Nakanishi T.: "Influence of cultivation temperature and viral infection of RTG-2 cells on the expression of rainbow trout classical MHC class I molecule"Dev.Comp.Immunol.. 24. 77 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Ototake M, Okamura K,Hashimoto K,Nakanishi T.: "The function of the most primitive MHC class I"Dev.Comp.Immunol.. 24. 79 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 中西照幸: "家畜ゲノム解析と新たな家畜育種戦略(畜産技術協会)"魚類における疾病抵抗性の遺伝と抵抗性系統の作出. 359-362 (2000)

    • Related Report
      2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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