Bioactivity and secretion of interleukin-18 (IL-18) generated by equine and feline IL-18 expression constructs.
Interleukin 18 (IL-18) is a cytokine capable of induction of IFNgamma, granulocyte monocyte-colony stimulating factor (GM-CSF), TNFalpha and IL-1 in immunocompetent cells. Equine and feline plasmid vectors expressing pro-IL-18, mature IL-18 and IL-18 fused to a synthetic signal sequence from human IL-1beta receptor antagonist protein (ILRAP), ILRAP-IL-18, have been generated. In vitro protein expression of these constructs was compared by Western blot analysis. These data demonstrated that ILRAP-IL-18 protein was secreted readily from transfected chinese hamster ovary (CHO) cells. A simple bioassay for human IL-18 was recently described using human myelomonocytic KG-1 cells, which produce human IFNgamma in response to human IL-18 in a dose dependent manner (Konishi et al., 1997). We demonstrated bioactivity of equine and feline IL-18 protein in transfection products of CHO cells using this assay. Bioactivity of ILRAP-IL-18 protein was demonstrated in the culture medium of transfected CHO cells. These data imply that the ILRAP-IL-18 construct shows potential for use in vivo, where cell secretion of protein is crucial.
O'Donovan
L H
LH
Division of Infection and Immunity, Institute of Comparative Medicine, University of Glasgow, 464 Bearsden Road, Garscube Estate, Glasgow G61 1QH, UK. l.o'donovan@bio.gla.ac.uk
McMonagle
E L
EL
Taylor
S
S
Argyle
D J
DJ
Nicolson
L
L
D000818:Animals; D001681:Biological Assay; D016466:CHO Cells; D002415:Cats / Q000276:immunology*; D006224:Cricetinae; D003412:Cricetulus; D005786:Gene Expression Regulation / Q000276:immunology*; D006736:Horses / Q000276:immunology*; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000096:biosynthesis; D053590:Interleukin 1 Receptor Antagonist Protein; D020382:Interleukin-18 / Q000032:analysis / Q000096:biosynthesis*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012795:Sialoglycoproteins / Q000235:genetics
C507763:IL1RN protein, human; D053590:Interleukin 1 Receptor Antagonist Protein; D020382:Interleukin-18; D011993:Recombinant Fusion Proteins; D012795:Sialoglycoproteins; D007371:Interferon-gamma