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Development of cytotoxicity test for outbred animals using recombinant viruses.

Research Project

Project/Area Number 05556053
Research Category

Grant-in-Aid for Developmental Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field Applied veterinary science
Research InstitutionThe University of Tokyo

Principal Investigator

KAI Chieko  Univ.of Tokyo, Fac.of Agr.Associate Professor, 農学部, 助教授 (10167330)

Co-Investigator(Kenkyū-buntansha) AIDA Yohko  Insti.of Phys.and Chem.Res.Researcher, 先任研究員 (50182994)
IHARA Takeshi  Nippon Inst for Biological Sci.Researcher, 研究員 (70150109)
YAMANOUCHI Kazuya  Nippon Inst.for Biological Sci.Chief Researcher, 主任研究員 (30072888)
FUMIO Kobune  National Inst.of Health Chief Researcher, 主任研究官 (80142644)
SAITOH Izumi  Institute of Medical Science Associate Professor, 医科学研究所, 助教授 (70158913)
遠矢 幸伸  東京大学, 農学部, 助手 (20180119)
Project Period (FY) 1993 – 1995
Project Status Completed (Fiscal Year 1995)
Budget Amount *help
¥13,200,000 (Direct Cost: ¥13,200,000)
Fiscal Year 1995: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1994: ¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1993: ¥7,000,000 (Direct Cost: ¥7,000,000)
KeywordsCTL / cell mediated cytotoxic assay / adenovirus vector / rinderpest virus / measles virus / bovine leukemia virus / recombinant / domestic animals / アデノウイルス / T細胞 / CAG / EF1α / 遺伝子組換え / 膜蛋白遺伝子 / 細胞傷害性T細胞 / lacZ遺伝子 / SRα
Research Abstract

The cytotoxic T lymphocytes (CTL) have been considered to play an important role in immunity against the infections of many kinds of viruses. However, the function of CTL in animals including domestic, companion or experimental animals except mice were not well studied because of the lack of CTL assay methods for those animals. It was difficult to produce the useful target cells in those animal systems since target cells should be obtained from the same animal which is infected by viruses to match MHC with the effector cells, and since the target cells should express virus antigens on the cell surface without showing CPE until they meet the effector cells.
The final aim of this project is to develop and establish the new cytotoxic T cell assay system which is useful for any kinds of animals by using improved adenovirus vectors. For developing the new assay system, we tested and obtained the optimal conditions in each subjects as follows.
(1) Host animal and cell ranges of adenovirus infection.
(2) The best promoters to express virus antigens in the adenovirus vector.
(3) Cloning and characterizations of virus genes whose products are considered to be the target of cellular cytotoxicity.
(4) Construction of recombinant adenoviruses with virus genes.
(5) The conditions to produce the target cells from peripheral lymphocytes of each animals after the infection of recombinant adenoviruses.
Since we tested several kinds of viruses, not all those conditions have been obtained in each system. But we succeeded to construct recombinant adenoviruses with measles and renderpest virus genes, and obtained the optimal conditions for producing the target cells from monkey lymphocytes infected with the recombinant adenoviruses. In addition, we identified the best promoters in adenovirus vectors, and the unique MHC molecules on bovine lymphocytes. These results are considered to contribute to the basic and applied studies for cellular cytotoxicity of animals using adenovirus vectors.

Report

(4 results)
  • 1995 Annual Research Report   Final Research Report Summary
  • 1994 Annual Research Report
  • 1993 Annual Research Report
  • Research Products

    (36 results)

All Other

All Publications (36 results)

  • [Publications] Shin, Y.-S. et al: "Detection of canine distemper virus nucleocapsid protein gene in canine peripheral blood mononuclear cells by RT-PCR." J. Vet. Med. Sci.57. 439-445 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Gemma,T. et al: "Suppression of canine parvovirus growth in CRFK cells by canine distemper virus." J. Vet. Med. Sci.57. 535-537 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Iwatsuki, K. et al.: "Immunohistochemical analysis of the lymphoid organs of dogs naturally infected with canine distemper virus." J. Comp. Path.113. 185-190 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Gemma,T. et al.: "Serological survey of canine distemper virus infection using enzyme-linked immunosorbent assay." J. Vet. Med. Sci.57. 761-763 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Okita, M. et al.: "Immunohistochemical studies of lymphoid tissues of rabbits infected with rinderpest virus." J. Comp. Path.112. 41-51 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Mamaev, L. V. et al.: "Characterisation of morbilliviruses isolated from Lake Baikal seals (Phoca sibirica)." Veterinary Microbiology. 44. 251-259 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 甲斐知恵子: "獣医微生物学pp.139-140, 207-210, 247-249(分担執筆)" 文永堂出版, 426 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 甲斐知恵子: "獣医伝染病学(第四版)pp. 79-80, 239-241(分担執筆)" 近代出版, 332 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Shin, Y.-S., al: "Detection of canine distemper virus nucleocapsid protein gene in canine peripheral blood mononuclear cells by RT-PCR." J.Vet.Med.Sci. 57. 439-445 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Gemma, T.ey al: "Suppression of canine parvovirus growth in CRFK cells by canine distemper virus." J.Vet.Med.Sci. 57. 535-537 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Iwatsuki, K.et al: "Immunohistochemical analysis of the lymphoid organs of dogs naturally infected with canine distemper virus." J.Comp.Path. 113. 185-190 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Gemma, T.et al: "Serological survey of canine distemper virus infection using enzyme-linked immunosorbent assay." J.Vet.Med.Sci. 57. 761-763 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Okita, M.et al: "Immunohistochemical studies of lymphoid tissues of rabbits infected with rinderpest virus." J.Comp.Path. 112. 41-51 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Mamaev., L.V.et al: "Characterisation of morbilliviruses isolated from Lake Baikal seals (Phoca sibirica)." Veterinary Microbiology. 44. 251-259 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Okita, M., et. al.: "Immunohistochemical studies of lymphoid tissues of rabbits infected with rinderpest virus." J. Comp. Path.112. 41-51 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Aida, Y., et. al.: "Identification of a new bovine MHC class II DRB allele by nucleotid sequencing and an analysis of phylogenetic relationship." Biochem. Biophys. Res. Commun.209. 981-988 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Fujita, A., et. al.: "Gene targeting with a replication-defective adenovirus vector." J. Virol.69. 6180-6190 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Iwatsuki, K., et. al.: "Immunohistochemical analysis of the lymphoid organs of dogs naturally infected with canine distemper virus." J. Comp. Path.113. 185-190 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Miyake, S., et. al.: "Efficient generation of recombinant adenoviruses using adenovirus DNA-terminal protein complex and a cosmid containing full-length adenovirus genome." Proc. Natl. Acad. Sci. USA. (in press).

    • Related Report
      1995 Annual Research Report
  • [Publications] Kai, C., et. al.: "Nucleotide sequences of the M gene of prevailing wild measles viruses and a comparison with subacute sclerosing panencephalitis virus." Virus Genes. (in press).

    • Related Report
      1995 Annual Research Report
  • [Publications] 甲斐知恵子: "獣医伝染病学(第四版)pp.79-80, pp.239-241. (分担執筆)" 近代出版, 426 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 甲斐知恵子: "獣医微生物学 pp.139-140, pp.207-210, pp.247-249.(分担執筆)" 文永堂出版, 332 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Maeda,K.et al.: "A gD homologous gene of feline herpesvirus type l encodes a hemagglutinin(gp60)." Virology. 202. 1034-1038 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Oshikamo,R.et.al.: "The molecular cloning and sequence of an open reading frame encoding for non-structural proteins of feline calicivirus F4 strain isolated in Japan." J.Vet.Med.Sci.56. 1093-1099 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Aida,et.al.: "Cloning of cDNAs and the molecular evolution of a bovine MHC class II DRA gene." Biochem.Biophys.Res.Commun.204. 195-202 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Xuan,X.et.al.: "Characterization of pseudorabies virus glycoprotein gII expressed by recombinant baculovirus." Virus Research. (In press). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Harada,T.et.al.: "Human liver cell line constitutively expressing nonstruatural proteins of hepatitis C virus." J.Gen.Virol.(In press). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Harada,S.et.al.: "Establishment of a cell line constitutively expressing E2 glycoprotein of hepatitis C virus and humoral responce of hepatitis C patients to the expressed protein." J.Gen.Virol.(In press). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] 甲斐知恵子: "獣医畜産新報" 分永堂出版, 4 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 甲斐知恵子: "科学" 岩波書店, 8 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Kamata,H.et al.: "Expression in baculovirus vector system of the nucleocapsid protein gene of rinderpest virus." J.Virol.Meth.43. 159-166 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Maeda K.et al.: "Identification and uncleotide sequence of a gene in feline herpesvirus type 1 homologous to the herpes simplex virus gene encoding the glycoprotein H." Archives of Virology. 132. 183-191 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Okita,M.et al.: "Comparison of the pathogenicity of rinderpest virus in defferent strains of rabbits." J.Vet.Med.Sci.55. 951-954 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Shin,Y-S.et al.: "Antigenic analysis of feline calicivirus capsid precursor protein and its deleted polypeptides produced in a mammalian cDNA expression system." Virus Research. 30. 17-36 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Takahashi,H.et al.: "Mechanism of pruritus and peracute death in mice induced by pseudorabies viurs(PRV)infection." J.Vet.Med.Sci.55. 913-920 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Toyosaki,T.et al.: "Localization of the viral antigen of feline immunodeficiency virus in the lymph nodes of cats at the early stage of infection." Archives of Virology. 131. 335-347 (1993)

    • Related Report
      1993 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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