Expression of biologically active human granulocyte-macrophage colony-stimulating factor in the silkworm (Bombyx...

Using virus derived from Bombyx mori (silkworm) nuclear polyhedrosis virus (BmNPV), we constructed an infectious recombinant virus carrying the human granulocyte-macrophage colony-stimulating factor (hGM-CSF) cDNA placed downstream from the polyhedrin promoter. The BmN cells infected with the recombinant baculovirus BmNPV-GM-CSF expressed recombinant (r)hGM-CSF up to 3.9 x 10(5) colony-formation units/ml in the medium. The maximum activity of rhGM-CSF was 4.3 x 10(6) colony-formation units/ml in the haemolymph of silkworm (Bombyx mori) larvae infected with BmNPV-GM-CSF. Three distinct species of GM-CSF of molecular masses 15, 18 and 20 kDa were detected by immunoblotting. The specific activity of rhGM-CSF purified from the haemolymph was up to 2.92 x 10(7) colony-formation units/mg. The isoelectric point of the purified rhGM-CSF was 5.5-5.7.