| RRC ID |
39316
|
| 著者 |
Ishihara K, Satoh I, Nittoh T, Kanaya T, Okazaki H, Suzuki T, Koyama T, Sakamoto T, Ide T, Ohuchi K.
|
| タイトル |
Preparation of recombinant rat interleukin-5 by baculovirus expression system and analysis of its biological activities.
|
| ジャーナル |
Biochim Biophys Acta
|
| Abstract |
Rat interleukin-5 (IL-5) cDNA was subcloned from peritoneal cells collected 4 h after intraperitoneal injection of Ascaris suum antigen solution into the immunized rats. Cysteine proteinase-deleted (CPd) rat IL-5 recombinant virus was constructed by inserting rat IL-5 cDNA into CPd virus having a deletion in the cysteine proteinase gene of the silkworm Bombyx mori nuclear polyhedrosis virus. On infection with the CPd rat IL-5 recombinant virus, the silkworm B. mori larvae produced rat IL-5 as a dimeric form in hemolymph. Recombinant rat IL-5 was purified more than 95.5% by anion-exchange chromatography and hydrophobic chromatography. The purified recombinant rat IL-5 promoted the proliferation of T88-M cells in a concentration-dependent manner, and its effect was inhibited by an anti-murine IL-5 neutralizing polyclonal antibody. When bone marrow cells from normal rats were incubated with recombinant rat IL-5 in medium containing methylcellulose, the colony formation by eosinophilic cells was induced. Furthermore, when rat peritoneal eosinophils were incubated with recombinant rat IL-5, the spontaneous decrease in the eosinophil viability was inhibited in time- and concentration-dependent manners. In addition, the recombinant rat IL-5-induced eosinophil survival was inhibited by an anti-murine IL-5 neutralizing polyclonal antibody. These findings suggest that rat IL-5 acts as B-cell growth factor II (BCGF-II), eosinophil differentiation factor (EDF), and eosinophil survival-enhancing factor.
|
| 巻・号 |
1451(1)
|
| ページ |
48-58
|
| 公開日 |
1999-8-12
|
| DOI |
10.1016/s0167-4889(99)00090-7
|
| PII |
S0167-4889(99)00090-7
|
| PMID |
10446387
|
| MeSH |
Animals
B-Lymphocytes / drug effects
Baculoviridae / genetics
Blotting, Western
Bombyx / metabolism
Cell Differentiation / drug effects
Cell Division / drug effects
Cell Line
Cell Survival / drug effects
Cloning, Molecular
Colony-Forming Units Assay
Electrophoresis, Polyacrylamide Gel
Eosinophils / drug effects
Hemolymph / metabolism
Interleukin-5 / genetics
Interleukin-5 / isolation & purification
Interleukin-5 / metabolism*
Rats
Recombinant Proteins / metabolism
|
| IF |
3.411
|
| 引用数 |
13
|
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
CELL BIOLOGY
|
| リソース情報 |
| ヒト・動物細胞 |
|
