Grant-in-Aid for Scientific Research (B).
|Research Institution||Oita Medical University|
YAMAMOTO Shunsuke Oita Medical Univ.,Pathology,Professor, 医学部, 教授 (90040188)
YOSHIDA Seiji Oita Medical Univ.,Pathology,Assistant, 医学部, 助手 (70182764)
AKIZUKI Shinishiro Oita Medical Univ.,Pathology,Assistant, 医学部, 助手 (80159334)
SETOGUCHI Mihoko Oita Medical Univ.,Pathology,Assistant, 医学部, 助手 (20236110)
樋口 安典 大分医科大学, 医学部, 助教授 (60040284)
|Project Fiscal Year
1990 – 1992
Completed(Fiscal Year 1992)
|Budget Amount *help
¥5,600,000 (Direct Cost : ¥5,600,000)
Fiscal Year 1992 : ¥300,000 (Direct Cost : ¥300,000)
Fiscal Year 1991 : ¥700,000 (Direct Cost : ¥700,000)
Fiscal Year 1990 : ¥4,600,000 (Direct Cost : ¥4,600,000)
|Keywords||CD14 / MS2 / osteopontin / cDNA / genome / expression / recombinant / mouse and human / オステオポンチン / マクロファ-ジ遺伝子 / マクロファ-ジcDNA / マクロファ-ジ / 膜蛋白 / 分泌蛋白 / Sppー1|
1.MS2:(1). Complete nucleotide sequences of the MS2 gene and its 5 upstream region were determined. (2). cis-regulatory element for MS2 expression in macrophages (Mphi) was investigated. (3). Significant homology of MS2 with hemorrhagic snake venom proteins were found. (4). Recombinant proteins for MS2 proteins (rMS2) and poly clonal and monoclonal antibodies for rMS2 (anti- rMS2) were produced. (5). PAP and FACS analysis revealed that MS2 was expressed in Mphi and neutrophils but not in lymphocytes and mast cells. (6). cDNA for the human homologue of MS2 was separated.
2. CD14:(1). The 5'upstream nucleotide sequences of the mouse and human CD14 genes were determined. (2). Promoter and enhancer regions for the mouse CD14 gene were defined. (3). Mouse rCD14 and polyclonal and monoclonal antibodies against rCD14 (anti- rCD14) were produced. PAP analysis using anti-rCD14 demonstrated CD14 in myelomonocytic cells but not in lymphocytes and mast cells. (5). TNFalpha production and LPS-induced shock were significantly suppressed with anti-rCD14.
3. OP:(1). The 5'upstream nucleotide sequences of the mouse and human OP genes were determined. (2). Complete nucleotide sequence of the human OP gene was determined. (3). Cis regulatory sequences of the mouse and human OP genes were determined. (4). rOP was produced using E. coli and vaculovirus vector systems. (5). rOP significantly suppressed haptotaxis of Mphi and melanoma (B16-BL) cells, and lung metastasis of B16-BL cells. (5). Antibodyagainst rOP was produced.