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1987 Fiscal Year Final Research Report Summary

Gene cloning and characterization of new HLA class II antigens

Research Project

Project/Area Number 60490021
Research Category

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

Allocation TypeSingle-year Grants
Research Field 広領域
Research InstitutionTokai University Shcool of Medicine

Principal Investigator

NAKATSUJI Takako  Tokai University School of Medicine, Instructor, 医学部, 助手 (00172346)

Co-Investigator(Kenkyū-buntansha) ANDO Asako  Tokai University School of Medicine, Instructor, 医学部, 助手 (40101935)
TSUJI Kimiyoshi  Tokai University School of Medicine, Professor, 医学部, 教授 (30055834)
Project Period (FY) 1985 – 1987
KeywordsHLA / Class II antigen / Gene cloning / Transfection / HLA-DO Antigen / 塩基配列
Research Abstract

1. A cDNA clone encoding a new HLA class II antigen <alpha> chain, named DO<alpha>, was isolated from a human CDNA library using the DR<alpha>, DO<alpha> and DP<alpha> cDNA probes.
The nucleotide sequence of the DO<alpha> cDNA was distinct from those of the DR<alpha>, the DO<alpha>, and the DP<alpha> cDNA, but showed some characteristic features of the class II antigen <alpha>-chain. We also isolated and identified genomic clones specifying the DO<alpha> gene. Genomic Southern hybridization analyses of cell lines with different HLA-DR serotypes indicated the existence of a single DO<alpha> gene that exhibited little restriction enzyme polymorphism.
2. To describe the function of gene products from this new DO locus, DO gene clone was introduced. The resultant transfectant cells were analyzed their expression and function using biochemical and cellular-immunological procedure.
3. A genomic clone specifying a new HLA class II antigen <beta> chain pseudogene, named DV<beta>, was isolated from a human genomic phage library using a DO<beta> cDNA probe.
Southern hybridization and nucleotide sequence analyses identified the <beta>2 domain exon (exon 3) with several deleterious mutations and the CP-TM-CY exon (connecting peptide, transmembrane, and cytoplasmic exon, exon 4), but the first, second and fifth exons of normal <beta> chains were entirely missing. The nucleotide sequence of these two exons were distinct from those of other class II <beta>-chain genes, but slightly more related to the DQ<beta> and the DX<beta> genes than other class II genes. Cosmid clone analysis around the DV<beta> gene placed DV<beta> between DX<alpha> and DQ<beta>, 15 kb upstream from DX<alpha>, which was consistence with that around DX<alpha> which was hown by RFLP(restriction fragment length polymorphism studies).

  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] Inoko, H.;Ando, A.;Kimura, M.;Tsuju, K.: J. Immunol.135. 2156-2159 (1985)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nakatsuji, T.;Inoko, H.;Ando, A.;Sato, T.;Koide, Y.;Tadakuma, T.;Yoshida, T. O.;Tsuji, K.: Immunogenetics. 25. 1-6 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Inoko, H.;Ando, A.;Ito, M.;Tsuji, K.: Human Immunol.16. 304-313 (1986)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ando, A.;Inoko, H.;Kimura, M.;Ogata, S.;Tsuji, K.: Human Immunol.17. 355-367 (1986)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ando, A.;Kawai, J.;Maeda, M.;Tsuji, K.;Trowsdale, J.;Inoko, H.: J. Immunol.submitted.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ando, A.;Inoko, H.;Nakatsuji, T.;Sato, T.;Awataguchi, S.;Tsuji, K.: "HLA in Asia-Oceania 1986" Aizawa, M. (Hokkaido University Press), 7 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Inoko.H., Ando, A., Kimura, M. & Tsuji, K.: "Isolation and characterization of the cDNA clone and genomic clones of a new HLA class II antigen heavy chain, DO<alpha>." J. Immunol.135. 2156-2159 (1985)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakatsuji,T., Inoko,H., Ando,A., Sato,T., Koide,Y., Tadakuma,T., Yoshida,T.O. & Tsuji,K.: "The role of transfected HLA-DQgenes in the mixed lymphocyte reaction like condition" Immunogenetics. 25. 1-6 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Inoko,H., Ando,A., Ito,M, & Tsuji,K.: "Southern hybridization analysis of DNA polymorphism in the HLA-D region" Human Immunology. 16. 304-313 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ando.A., Inoko,H., Kimura,M., Ogata,S & Tsuji,K.: "Isolation and allelic polymorphism of cDNA clones and genomic clones of HLA-DP heavy and light chains" Human Immunology. 17. 355-367 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Andods,A., Kawai,J., Maeda,M., Tsuji,K., Torowsdale,J. & Inoko,H.: "J. Immunol." Mapping and nucleotide sequence of a new HLA class II light chain gene, DV<beta>. submitted.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ando,A., Inoko,H., Nakatusji,T., Sato,T., Awataguchi,S., & Tsuji,K.: HLA in Asia-Oceania 1986 , (ed) Aizawa. M., (Hokkaido Univ. Press). Isolation and characterization of cDNA and genomic clones for new HLA class II antigen heavy and light chains, 859-865 (1987)

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

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Published: 1989-03-30  

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