PURPOSE: To obtain a human IL18-IL2 fusion protein by genetic engineering methods and investigate its antitumor activity and mechanism in tumor-bearing mouse models. METHODS: Human IL-18, IL-2 and the fusion gene IL18-IL2 were obtained by PCR, inserted into pBV220 vector and expressed in BL21 (DE3) by 42 °C heat induction. Purified proteins were analyzed by SDS-PAGE and Western blot. The biological activity of IL18-IL2 was first determined by its ability to augment IFN-γ production in PBMCs. Then tumor-bearing mouse models of mouse Lewis lung carcinoma (LLC), human large cell lung carcinoma (NCI-H460) and human colorectal carcinoma (HCT-116) were established to investigate the antitumor activity and mechanism of IL18-IL2. RESULTS: IL18-IL2 was confirmed by Western blot, and its molecular weight was about 34.5 kDa. IL18-IL2 could significantly enhance production of IFN-γ in PBMCs in vitro and induce significant tumor regression in tumor-bearing mouse models of LLC, NCI-H460 and HCT-116 than that of IL-18 and IL-2 separately or combination using. In the mice bearing HCT-116 and LLC, IFN-γ concentrations and natural killer cell cytotoxicity were highly enhanced by IL18-IL2. Anti asialo GM1 could reduce natural killer cell cytotoxicity, production of IFN-γ, and regression of LLC tumor aroused by IL18-IL2. CONCLUSIONS: These results suggested the IL18-IL2 fusion protein showed a synergetic effect on tumor regression, which was related to the great ability of IL18-IL2 in enhancing IFN-γ production and natural killer cell cytotoxicity. The fusion protein was a potential antitumor reagent in cancer immunotherapy.
PURPOSE: To obtain a humanIL18-IL2 fusion protein by genetic engineering methods and investigate its antitumor activity and mechanism in tumor-bearing mouse models. METHODS:HumanIL-18, IL-2 and the fusion gene IL18-IL2 were obtained by PCR, inserted into pBV220 vector and expressed in BL21 (DE3) by 42 °C heat induction. Purified proteins were analyzed by SDS-PAGE and Western blot. The biological activity of IL18-IL2 was first determined by its ability to augment IFN-γ production in PBMCs. Then tumor-bearing mouse models of mouseLewis lung carcinoma (LLC), human large cell lung carcinoma (NCI-H460) and humancolorectal carcinoma (HCT-116) were established to investigate the antitumor activity and mechanism of IL18-IL2. RESULTS:IL18-IL2 was confirmed by Western blot, and its molecular weight was about 34.5 kDa. IL18-IL2 could significantly enhance production of IFN-γ in PBMCs in vitro and induce significant tumor regression in tumor-bearing mouse models of LLC, NCI-H460 and HCT-116 than that of IL-18 and IL-2 separately or combination using. In the mice bearing HCT-116 and LLC, IFN-γ concentrations and natural killer cell cytotoxicity were highly enhanced by IL18-IL2. Anti asialo GM1 could reduce natural killer cell cytotoxicity, production of IFN-γ, and regression of LLC tumor aroused by IL18-IL2. CONCLUSIONS: These results suggested the IL18-IL2 fusion protein showed a synergetic effect on tumor regression, which was related to the great ability of IL18-IL2 in enhancing IFN-γ production and natural killer cell cytotoxicity. The fusion protein was a potential antitumor reagent in cancer immunotherapy.
Authors: Y Gu; K Kuida; H Tsutsui; G Ku; K Hsiao; M A Fleming; N Hayashi; K Higashino; H Okamura; K Nakanishi; M Kurimoto; T Tanimoto; R A Flavell; V Sato; M W Harding; D J Livingston; M S Su Journal: Science Date: 1997-01-10 Impact factor: 47.728
Authors: Pushpa Saranya Kollipara; Do Hee Won; Chul Ju Hwang; Yu Yeon Jung; Heui Seoung Yoon; Mi Hee Park; Min Jong Song; Ho Sueb Song; Jin Tae Hong Journal: Biomol Ther (Seoul) Date: 2014-02 Impact factor: 4.634