[Expression and purification of murine interleukin 18 in Escherichia coli and its antitumor effects].
Total RNA was extracted from murine hepatocytes, and the cDNA of interleukin 18(IL-18) was amplified by RT-PCR. The cDNA was introduced into the expression vector pJW2 and sequenced. Under heat induction, the recombinant murine IL-18(rmIL-18) was expressed in inclusion bodies in E. coli with the yield accounting for 18% of total bacteria proteins. The inclusion bodies were dissolved with 5 mol/L urea, and rmIL-18 was purified using Sephadex G-100 column chromatography. In the presence of 0.5 mg/L Con A, the purified rmIL-18 showed dose-dependent IFN-gamma-inducing activity in murine splenocytes. The purified rmIL-18 exhibited significant antitumor effects in Kunming mice challenged intraperitoneally (i.p.) with H22 hepatocarcinoma when administered 10 micrograms rmIL-18 i.p. on days 1, 4 after challenge, and the mice survived resisted the rechallenged with H22 cells.
Zhou
H
H
Research Center for Biochemistry and Molecular Biology, Xuzhou Medical College, Xuzhou 221002, China.
Zhao
H R
HR
Pei
D S
DS
Lu
L
L
Hu
S Q
SQ
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002850:Chromatography, Gel / Q000379:methods; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D007371:Interferon-gamma / Q000378:metabolism; D020382:Interleukin-18 / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*; D008114:Liver Neoplasms, Experimental / Q000276:immunology / Q000473:pathology / Q000517:prevention & control; D051379:Mice; D009368:Neoplasm Transplantation; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology; D013154:Spleen / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D013997:Time Factors; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D018076:DNA, Complementary; D020382:Interleukin-18; D011994:Recombinant Proteins; D007371:Interferon-gamma