title: Structure and characterization of hamster IL-12 p35 and p40.
abstract: Complementary DNAs coding for two subunits of hamster interleukin-12 (IL-12), p35 and p40, were cloned from a hamster dendritic cell (DC) cDNA library. The cloning demonstrated that hamster IL-12 consisted of a p35 subunit with 216 amino acid (aa) residues and a p40 subunit with 327 aa. Structural comparison of hamster p35 and p40 at the protein level showed the highest homologies with each counterpart of sigmodon (hispid cotton rat). The gene expressions of hamster IL-12 p35 and p40 in bone marrow (BM) cells cultured in the presence of mouse granulocyte macrophage-colony-stimulating factor (mGM-CSF) and IL-4 were up-regulated during culture. Immunoblot analysis of 293 cells transfected with hamster p35 and p40 expression vectors suggested the presence of a covalently linked p35/p40 heterodimer. Furthermore, supernatant from the 293 cells transfected with both expression vectors induced the up-regulation of interferon-gamma (IFN-gamma) mRNA in hamster splenocytes, indicating that the p35/p40 heterodimer IL-12 protein present in the supernatant was functional. These results suggest that the vectors containing hamster IL-12 cDNA might be suitable tools for developing an immunotherapeutic approach against experimental cancer in a hamster model.
authors.affiliation: Growth Factor Division, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, 104-0045, Tokyo, Japan
authors.forename: Kouji
authors.initials: K
authors.lastname: Maruyama
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Takigawa
authors.forename: Yasuto
authors.initials: Y
authors.lastname: Akiyama
authors.forename: Takashi
authors.initials: T
authors.lastname: Hojo
authors.forename: Noriko
authors.initials: N
authors.lastname: Nara-Ashizawa
authors.forename: Jin-yan
authors.initials: JY
authors.lastname: Cheng
authors.forename: Morihiro
authors.initials: M
authors.lastname: Watanabe
authors.forename: Ken
authors.initials: K
authors.lastname: Yamaguchi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D006224:Cricetinae; D005260:Female; D007371:Interferon-gamma / Q000378:metabolism; D018664:Interleukin-12 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D053765:Interleukin-12 Subunit p35; D053762:Interleukin-12 Subunit p40; D008969:Molecular Sequence Data; D021122:Protein Subunits / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013154:Spleen / Q000378:metabolism
chemical_list: D053765:Interleukin-12 Subunit p35; D053762:Interleukin-12 Subunit p40; D021122:Protein Subunits; D018664:Interleukin-12; D007371:Interferon-gamma