K. Ishihara et al., Preparation of recombinant rat interleukin-5 by baculovirus expression system and analysis of its biological activities, BBA-MOL CEL, 1451(1), 1999, pp. 48-58
Citations number
53
Categorie Soggetti
Cell & Developmental Biology
Journal title
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH
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 de
letion in the cysteine proteinase gene of the silkworm Bombyx mori nuclear
polyhedrosis virus. On infection with the CPd rat IL-5 recombinant virus, t
he 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 chrom
atography and hydrophobic chromatography. The purified recombinant rat IL-5
promoted the proliferation of T88-M cells in a concentration-dependent man
ner, and its effect was inhibited by an anti-murine IL-5 neutralizing polyc
lonal antibody. When bone marrow cells from normal rats were incubated with
recombinant rat IL-5 in medium containing methylcellulose, the colony form
ation by eosinophilic cells was induced. Furthermore, when rat peritoneal e
osinophils were incubated with recombinant rat IL-5, the spontaneous decrea
se in the eosinophil viability was inhibited in time- and concentration-dep
endent manners. In addition, the recombinant rat IL-5-induced eosinophil su
rvival was inhibited by an anti-murine IL-5 neutralizing polyclonal antibod
y. These findings suggest that rat IL-5 acts as B-cell growth factor II (BC
GF-II), eosinophil differentiation factor (EDF), and eosinophil survival-en
hancing factor. (C) 1999 Elsevier Science B.V. All rights reserved.