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2000 Fiscal Year Final Research Report Summary

Study on the structure and function of viral proteins using artificial particles of human rotavirus

Research Project

Project/Area Number 10470080
Research Category

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

Allocation TypeSingle-year Grants
Section一般
Research Field Virology
Research InstitutionFUJITA HEALTH UNIVERSITY

Principal Investigator

TANIGUCHI Koki  Fujita Health University, Dept.Virol.& Parasitol, Professor, 医学部, 教授 (40094213)

Co-Investigator(Kenkyū-buntansha) TAKEDA Naokazu  National Institute of Infectious Diseases, Virology II, Head, ウイルスII部, 室長 (90132894)
KOBAYASHI Nobumichi  Sapporo Med.Univ., Dept.of Hygiene, Associate Professor, 医学部, 助教授 (80186759)
URASAWA Tomoko  Sapporo Med.Univ., Dept.Health Allied Professions, Professor, 保健医療学部, 教授 (90045378)
MATSUURA Yoshiharu  Osaka Univ., Res.Institute Microbial Dis., Professor, 微生物病研究所, 教授 (50157252)
Project Period (FY) 1998 – 2000
KeywordsROTAVIRUS / BACULOVIRUS / PROTECTION / VACCINE / ARTIFICIAL PARTICLE / MUCOSAL ADJUVANT / CTL / REVERSE GENETICS
Research Abstract

1. Self-assembled, artificial and empty single-shelled particles (VP2/6) were prepared by using baculovirus expression system, and they were purified in a large quantity. VP2/6 particles accompanied with mucosal adjuvant (mLT-135, mLT-H44A, recombinant B subunit) were intranasally immunized in mice. Significant IgG and IgA responses in serum and stool were observed when mLT-135 was used as a mucosal adjuvant. In the inoculation with viruent murine strain EW for the immunized mice, a marked decrease of virus antigen in stools was observed in the case of mLT-135. Furthermore, a strong CTL response was also observed. Expression of mRNA for Th1 or Th2 cytokines was detected in the spleen cells from the immunized mice. Thus, artificial empty single-shelled particles combined with mLT-135 induced efficiently both of humoral and cellular immune responses in mice, suggesting the utility of the artificial particles and mucosal adjuvant as a vaccine for humans against rotavirus infection.
2. We o … More bserved that human rotaviruses with G serotype other than G3 also induced diarrhea in suckling mice. This finding is useful for infection experiments using human rotavirus strains in mice.
3. We carried out the following experiments for trying to establish reverse genetics of rotavirus.(1) RNA transcripts from VP4 or VP7 gene of strain SAT11-L2 which is less dependent on trypsin were transfected to MA 104 cells with the aid of a helper virus, human strain KU which is highly dependent on trypsin.(2) We prepared several constructs for transfection : T7 promoter-rotavirus VP4 or VP7 gene-rybozyme from type D hepatitis virus-T7 terminater. Truncated NSP1 genes which are not necessarily required for replication were connected to picornavirus IRES and GFP gene, and they were inserted in the above plasmid. The RNA transcripts were transfected into MA 104 cells infected with human strain KU.The screening assay for detecting the cells expressing GFP or producing infectious reassortant virus was developed. However, infectious artificial rotavirus has not been successfuly isolated so far. Less

  • Research Products

    (24 results)

All Other

All Publications (24 results)

  • [Publications] M.I.Adah: "First detection of human group C rotaviruses in Nigeria and sequence analysis of their genes encoding VP4, VP6 and VP7 proteins."J.Med.Virol.. (in press). (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] N.Kobayashi: "Sequence analysis of structural and non structural proteins of a human group B rotavirus detected in Calcutta, India."J.Med.Virol.. (in press). (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Takahashi: "Analysis of anti-rotavirus activity of extract from Stevia rebaudiana."Antiviral Res.. 49. 15-24 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Kojima: "Rearrangements generated in double genes, NSP1 and NSP3, of viable progenies from a human rotavirus strain."Virus Res.. 67. 163-171 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] J.Okada: "New P serotype of group A human rotavirus closely related to that of a porcine rotavirus."J.Med.Virol.. 60. 63-69 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] MU Ahmed: "Analysis of human rotavirus G serotype in Bangladesh by enzyme-linked immunosorbent assay and polymerase chain reaction."J.Diarrheal Dis.Res. 17. 22-27 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] C.Zao: "Sequence analysis of VP1 and VP7 genes suggests occurrence of a reassortment of G2 rotavirus responsible for an epidemic of gastroenteritis."J.Gen.Virol.. 80. 1407-1415 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] J.Okada: "Functional analysis of the heterologous NSP1 genes in the genetic background of simian rotavirus SA11."Arch.Virol.. 144. 345-353 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] J.Okada: "Analysis on reassortment of rotavirus NSP1 genes lacking coding region for cysteine-rich zinc finger motif."Arch.Virol.. 144. 1439-1449 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M.Ramachandran: "Detection and characterization of novel rotavirus strains in the United States."J.Clin.Microbiol. 36. 3223-3229 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] H.Wu: "Serological and genomic characterizations of human rotaviruses detected in China."J.Med.Virol.. 55. 168-176 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] J.Okada: "Preferential selection of heterologous G3-VP7 gene in the genetic background of simian rotavirus SA11 detected by using a homotypic single-VP7 gene-substitution reassortant."Antiviral Res.. 42. 712-720 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M.Ramachandran et al.: "Detection and characterization of novel rotavirus strains in the United States."J.Clin.Microbiol.. 36. 3223-3229 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Wu et al.: "Serological and genomic characterizations of human rotaviruses detected in China."J.Med.Virol.. 55. 168-176 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] J.Okada et al: "Preferential selection of heterologous G3-VP7 gene in the genetic background of simian rotavirus SA11 detected by using a homotypic single-VP7 gene-substitution reassortant."Antiviral Res.. 38. 15-24 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] C.Zao, W.et al.: "Sequence analysis of VP1 and VP7 genes suggests occurrence of a reassortment of G2 rotavirus responsible for an epidemic of gastroenteritis."J.Gen.Virol.. 80. 1407-1415 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ahmed M.U.et al.: "Analysis of human rotavirus G serotype in Bangladesh by enzyme-linked immunosorbent assay and polymerase chain reaction."J.Diarrheal Dis.Res.. 17. 22-27 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] J.Okada et al.: "Analysis on reassortment of rotavirus NSP1 genes lacking coding region for cysteine-rich zinc finger motif."Arch.Virol.. 144. 345-353 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] J.Okada et al.: "Functional analysis of the heterologous NSP1 genes in the genetic background of simian rotavirus SA11."Arch.Virol.. 144. 1439-1449 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] J.Okada et al.: "New P serotype of group A human rotavirus closely related to that of a porcine rotavirus."J.Med.Virol.. 60. 63-69 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Kojima et al.: "Rearrangement generated in double genes, NSP1 and NSP3, of viable progenies from a human rotavirus strain."Virus Res.. 67. 163-171 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takahashi K.et al.: "Analysis of anti-rotavirus activity of extract from Stevia rebaudiana."Antiviral Res.. 49. 15-24 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] N.Kobayashi et al.: "Sequence analysis of structural and non structural proteins of a human group B rotavirus detected in Calcutta, India."J.Med.Virol.. (in press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.I.Adah et al.: "First detection of human group C rotaviruses in Nigeria and sequence analysis of their genes encoding VP4, VP6 and VP7 proteins."J.Med.Virol.. (in press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2002-03-26  

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