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Abstract Using sitedirected mutagenesis and DNA recombinant technology, GGC fragment was inserted into human IL-18 cDNA. The mutated IL-18 cDNA was constructed into plasmid pPIC9K, then transformed into Pichia pastoris GS115 and efficiently expressed. A Glycine residue was inserted into the recombinant IL-18 between Arg39 and Asp40 to form a RGD motif. The mutated IL18 was termed IL-18-RGD. The protein was purified with Sephadex G100. The cell culture of melanoma B16 showed that IL-18-RGD efficiently inhibited B16 tumor growth, IC50 = 8.10μmol/L, but the inhibitory effect of IL-18 was not detected. Both IL-18-RGD and IL-18 showed inhibitory activities on mice loaded B16 in vivo, the inhibitory efffct of IL-18-RGD was stronger than that of IL-18. The inhibitory activities of IL-18-RGD and IL-18 on bFGF induced angiogenesis on chorioallantoic membranes were detected. There was no differences between IL-18-RGD and wild IL-18 in the activity of inducing IFN-γ in human PBMC.
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Received: 08 February 2006
Published: 25 February 2006
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