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Developmental biotechnological and molecular immunological analysis of host defense mechanisms against protozoan infection

Research Project

Project/Area Number 11460144
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied veterinary science
Research InstitutionObihiro University of Agriculture and Veterinary Medicine

Principal Investigator

NAGASAWA Hideyuki  Obihiro University of Agriculture and Veterinary Medicine, National Research Center for Protozoan Diseases, Professor, 原虫病研究センター, 教授 (60172524)

Co-Investigator(Kenkyū-buntansha) TOYODA Yutaka  Obihiro University of Agriculture and Veterinary Medicine, National Research Center for Protozoan Diseases, Emiritas Professor, 畜産学部, 名誉教授 (90050418)
IGARASHI Ikuo  Obihiro University of Agriculture and Veterinary Medicine, National Research Center for Protozoan Diseases, Professor, 畜産学部, 教授 (80159582)
FUJISAKI Kozo  Obihiro University of Agriculture and Veterinary Medicine, National Research Center for Protozoan Diseases, Professor, 畜産学部, 教授 (00292095)
YOKOYAMA Minesuke  Research Center of Mitsubishi Life Science, Chief Researcher, 室長
SUZUKI Hiroshi  Obihiro University of Agriculture and Veterinary Medicine, National Research Center for Protozoan Diseases, Professor, 畜産学部, 教授 (60333473)
Project Period (FY) 1999 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥8,200,000 (Direct Cost: ¥8,200,000)
Fiscal Year 2001: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2000: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1999: ¥3,600,000 (Direct Cost: ¥3,600,000)
Keywordsprotozoan diseases / Toxoplasma gondii / Neospora caninum / Cryptosporidium parvum / Transgenic / apoptosis / protective immunity / P0 protein
Research Abstract

Protozoan parasites are most important causative of infectious diseases in the world. The subjectives of this study have focused on the prevention of protozoan diseases through the studies on biological substances and products derived from protozoa themselves, as well as on host defense mechanisms. Genetically modified animals such as transgenic mice and knockout mice are known to be powerful tools for analysis of gene function in vivo.
(1) Identification of specific molecules from Toxoplasma gondii, Babesia sp., Trypanosoma sp., Neospora caninum, Cryptosoridium parvum using molecular biological techniques.
(2) Localization of immunogenic molecules from protozoan parasites using IFA, confocal leser microscope, immuno-EM.
(3) Analysis of host immune responses using knockout mice as well as transgenic mice such as SAG-1 transgenic mouse, IFN-γ knockout mouse, scavenger receptor (SR) knockout mouse and TNF-α knockout mouse.
SAG-1 (p30), the major surface protein of Toxoplasma gondii, is consi … More dered as an important ligand in the process of host cell invasion. To analyze the mechanism of host cell irivasion and immune response in toxoplasmosis, we have established transgenic mice carrying p30 for the first time in the world. Moreover, many kind of knockout mice deficient in cytokines or receptors have been used for analysis of the host defense mechanisms against protozoan infection in our research center.
It was found that among surface molecules of antigen presenting cells (APC), class II, CD80 and CD86 have a crucial role for protective immune responses against protozoan infection. Although expression level of CD28 on T cells was not changed after infection, effective molecules to activate T cells, that is class II, CD80 and CD86 on the APC, were significantly detectable with high expression. SAG-1 (p30) is considered as an important ligand in the process of host cell invasion. If this protein is produced by host cells and interfere with that of parasites, then the host will become resistant to parasite invasion. Or, it may become more susceptible to infection due to the lack of immune response against p30 antigen. We have obtained several P30-founder mice expressing the gene product and transmitting the gene to the next generation, by using F1 (C57BL/6J x C3H/He) embryos. As far as we are aware, this is the first transgenic mice bearing a protozoan gene derived from T. gondii. We found that P30 molecule is one of important trigger to introduce the host immune responses against Toxoplasma infection. Furthermore, apoptosis of host cells is one of escape mechanisms of protozoan parasite against host protective immunity. Less

Report

(4 results)
  • 2001 Annual Research Report   Final Research Report Summary
  • 2000 Annual Research Report
  • 1999 Annual Research Report
  • Research Products

    (30 results)

All Other

All Publications (30 results)

  • [Publications] Tanaka, T. et al.: "Expression of recombinant Toxoplasma gondii P24"J. Vet. Med. Sci.. 61(11). 1235-1239 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Xuan, X. et al.: "Characterization of P15 antigen of Cryptosporidium parvum expressed by a recombinant vaccinia virus"J. Protozool. Res.. 9(1). 32-40 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Seng, S. et al.: "Expression of SAG-1 of Toxoplasma gondii in transgenic mice"Parasitol. Res.. 86. 263-269 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Inoue, N. et al.: "Novel species specific antigens of Trypanosoma congolense and their different localization among life-cycle stages"J. Vet. Med. Sci.. 62(10). 1041-1045 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Makala, L.H.C.et al.: "Antigen presentation by murine peritoneal cavity macrophage-derived dendritic cells"Pathobiology. 69. 104-112 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Nishikawa, Y. et al.: "Interferon-gamma-induced apoptosis in host cells infected with Neospora caninum"Parasitology. 123. 25-31 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Tanaka, T., Xuan, X., Kato, M., Nagasawa, H., Fujisaki, K., Makimi, T. and Suzuki N.: "Expression of recombinant Toxoplasma gondii P24"J. Vet. Med. Sci.. 61(11). 1235-1239 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Xuan, X., Zhang, S., Kamio, T., Tsushima, Y., Kamada, T., Nishikawa, Y., Otsuka, H., Karanis, P., Igarashi, I., Nagasawa, H., Fujisaki, K. and Mikami, T.: "Characterization of P15 antigen of Cryptosporidium parvum expressed by a recombinant vaccinia virus."J. Protozool. Res.. 9(1). 32-40 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Seng, S., Yokoyama, M., Suzuki, R., Maki, Y. Kato, M., Lim, C., Zayatiin, B., Inoue, N., Xuan, X., Igarashi, I., Nagasawa, H., Fujisaki, K., Mikami, T., Suzuki, N. and Toyoda, Y.: "Expression of SAG-1 of Toxoplasma gondii in transgenic mice"Parasitol. Res.. 86. 263-269 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Inoue, N., Lluz, A.F., Mori, T., Nagasawa, H., Fujisaki, K., Mikami, T.: "Novel species specific antigens of Trypanosoma congolense and their different localization among life-cycle stages"J. Vet. Med. Sci.. 62(10). 1041-1045 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Makala, L.H.C., Nishikawa, Y., Kamada, T., Xuan, X., Nagasawa, H.: "Antigen presentation by murine peritonal cavity macrophage-derived dendritic cells"Pathobiology. 69. 104-112 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Nishikawa, Y., Mishima, M., Nagasawa, H., Igarashi, I., Fujisaki, K., Otsuka, H. and Mikami, T.: "Interferon-gamma-induced apoptosis in host cells infected with Neospora caninum"Parasitology. 123. 25-31 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Makala, L. H. C. et al.: "Comparison of the accessory activity of murine peritoneal cavity macrophage derived dendritic cells and peritoneal cavity macrophages in a mixed lymphocyte reaction"J. Vet. Med. Sci.. 63(12). 1271-1277 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Nishikawa, Y. et al.: "In the absence of endogenous gamma interferon, mice acutely infected with Neospora caninum succumb to a lethal immune response characterized by inactivation of peritoneal macrophages"Clin. Diagn. Lab. Immunol.. 8(4). 811-817 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Mishima, M. et al.: "Construction of recombinant feline herpesvirus type 1 expressing Toxoplasma gondii surface antigen 1"Mol. Biochem. Parasitol.. 117(2). 103-106 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Nishikawa, Y. et al.: "Expression of canine interferon-g by a recombinant vaccinia virus and its antiviral effect"Virus Res.. 75(2). 113-121 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Mishima, M. et al.: "Modified protection against Toxoplasma gondii lethal infection and brain cyst formation by vaccination with SAG2 and SRS1"J. Vet. Med. Sci.. 63(4). 433-438 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Makala, L. H. C. et al.: "Antigen presentation by murine peritoneal cavity macrophage-derived dendritic cells"Pathobiology. 69(2). 104-112 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Nishikawa,Y. et al.: "Delivery of Neospora caninum surface protein, NcSRS2 (Nc-p43), to mouse using recombinant vaccinia virus."Parasitol.Res.. 86. 934-939 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Nishikawa,Y. et al.: "Immunisation of dogs with a canine herpesvirus vector expressing Neospora caninum surface protein, NcSRS2."Int.J.Parasitol.. 30. 1167-1171 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Inoue,N. et al.: "Novel species specific antigens of Trypanosoma congolense and their different localization among life-cycle stages."J.Vet.Med.Sci.. 62. 1041-1045 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Igarashi,I. et al.: "Immunization with recombinant surface antigens p26 with Freund's adjuvants against Babesia rodhaini infection."J.Vet.Med.Sci.. 62. 713-723 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Ikadai,H. et al.: "Protein analysis of Babesia caballi merozoites by two-dimensional polyacrylamide gel electrophoresis and western blotting."J.Vet.Med.Sci.. 62. 323-327 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Seng,S. et al.: "Expression of SAG-1 of Toxoplasma gondii in transgenic mice."Parasitol.Res.. 86. 263-269 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Ikadai H.et al.: "Cloning and expression of a 48-kilodalton Babesia caballi merozoite rhoptry protein and potential use of the recombinant antigen in an enzyme-linked immunosorbent assay"J. Clin. Microbiol.. 37. 3475-3480 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Xuan X.et al.: "Characterization of P15 antigen of Cryptosporidium parvum expressed by a recombinant vaccinia virus"J. Protozool. Res.. 9. 32-40 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Tanaka T.et al.: "Expression of recombinant Toxoplasma gondii P24"J. Vet. Med. Sci.. 61. 1235-1239 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Tanaka T.et al.: "Expression of Babesia equi merozoite antigen-2 by recombinant baculovirus and its use in the ELISA"Int. J. Parasitol.. 29. 1803-1808 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Ikadai H.et al.: "Inhibitory effect of monoclonal antibodies on the growth of Babesia caballi"Int. J. Parasitol.. 29. 1785-1791 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Seng S.et al.: "Increased susceptibility to Toxoplasma gondii infection in SAG-1 transgenic mice"Int. J. Parasitol.. 29. 1433-1436 (1999)

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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