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Human IL18-IL2 fusion protein as a potential antitumor reagent by enhancing NK cell cytotoxicity and IFN-γ production

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Abstract

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.

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Acknowledgments

We gratefully acknowledge the generous support of these studies by the Youth Foundation of Yantai University (No. YX08Z3). We also thank Dr. Dong-Xiao Feng for critical review of the manuscript.

Conflict of interest

We declare that we have no any conflict of interest.

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Authors and Affiliations

  1. School of Pharmacy, Yantai University, Yantai, 264005, China

    Guangying Du, Guoying Zhang, Kun Liu & Jingwei Tian

  2. Non-Clinical Research Department, Luye Pharma Group Ltd., Yantai, 264003, Shandong, China

    Guangying Du, Liang Ye, Qiuju Dong & Jingwei Tian

Authors
  1. Guangying Du
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  2. Liang Ye
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  3. Guoying Zhang
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  4. Qiuju Dong
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  5. Kun Liu
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Correspondence to Jingwei Tian.

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Du, G., Ye, L., Zhang, G. et al. Human IL18-IL2 fusion protein as a potential antitumor reagent by enhancing NK cell cytotoxicity and IFN-γ production. J Cancer Res Clin Oncol 138, 1727–1736 (2012). https://doi.org/10.1007/s00432-012-1248-5

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  • DOI: https://doi.org/10.1007/s00432-012-1248-5

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