[Recombinant cIL-18-binding protein as an antagonist to cIL-18 enhanced PBMCs secreting IFN-gamma].
OBJECTIVE
Interleukin-18 (IL-18) is a proinflammatory cytokines that plays a key role of immune regulation through activating the Th 1 cell and NK cell secreting interferon-gamma (IFN-gamma). IL-18-binding proteins(IL-18BP) secreted in human and mouse can antagonise the activity of IL-18. The homologue gene of IL-18BP from fowlpox virus genome was predicted and identified of its expressed protein, to use as an antagonist to IL-18 guiding disease.


METHODS
The cIL-18BP gene was isolated from the genome of fowlpox vaccine virus by PCR through a pair of special primers and cloned into vectors pPICZalphaA, then expressed in yeast GS115. The biologic activity of recombinant proteins binding with cIL-18 was measured, inhibiting the activity of cIL-18 enhancing relative cells secreting IFN-gamma was confirmed through ELISA.


RESULTS
The cIL-18BP gene was cloned from fowlpox virus and acquired high performance expression in yeast systems induced with methanol identified by SDS-PAGE. Recombinant cIL-18BP purified can bind specifically with recombinant cIL-18 determined by ELISA test. cIL-18BP can antagonise the activity of cIL-18 with determining the IFN-gamma concentration secreting in PBMCs and MSB1 cells stimulated by cIL-18 respectively.


CONCLUSIONS
The experiment indicates that recombinant cIL-18BP can inhibit the activity of cIL-18 stimulating related immune cells secreting IFN-gamma. Deletion of the cIL-18BP from virus may improve the safety and immunogenicity of fowlpox virus vaccine.
Liu
Wenqiang
W
Agricultural College of Liaocheng University, Prevention and Control Technology Institute of Animal Disease in Liaocheng University, Liaocheng 252059, China. liuwqsky@sina.com
Liu
Guiqin
G
Qin
Xuling
X
Wang
Guiying
G
D000818:Animals; D018957:CD59 Antigens / Q000276:immunology*; D002352:Carrier Proteins; D011345:Fenofibrate / Q000037:antagonists & inhibitors*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism / Q000494:pharmacology*; D007371:Interferon-gamma / Q000037:antagonists & inhibitors*; D020382:Interleukin-18 / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D018957:CD59 Antigens; D002352:Carrier Proteins; D036341:Intercellular Signaling Peptides and Proteins; D020382:Interleukin-18; D011994:Recombinant Proteins; C118370:interleukin-18 binding protein; D007371:Interferon-gamma; D011345:Fenofibrate