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Preparation of recombinant rat interleukin-5 by baculovirus expression system and analysis of its biological activities1A part of this paper was presented at the Workshop for Eosinophils in Allergy and Related Diseases, held in Tokyo, Japan, June 20, 1998.1
- Source :
- Biochimica et Biophysica Acta (BBA) - Molecular Cell Research. (1):48-58
- Publisher :
- Elsevier Science B.V.
-
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.
- Subjects :
- Eosinophil survival-enhancing factor
Growth factor
medicine.medical_treatment
B-Cell growth factor II
Cell Biology
Biology
Eosinophil
biology.organism_classification
Recombinant virus
Molecular biology
law.invention
medicine.anatomical_structure
Antigen
law
Polyclonal antibodies
medicine
Recombinant DNA
biology.protein
Interleukin-5
Eosinophil differentiation factor
Molecular Biology
Interleukin 5
Ascaris suum
Subjects
Details
- Language :
- English
- ISSN :
- 01674889
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Biochimica et Biophysica Acta (BBA) - Molecular Cell Research
- Accession number :
- edsair.doi.dedup.....66592f4248ed913efac840ffa58d1cda
- Full Text :
- https://doi.org/10.1016/S0167-4889(99)00090-7