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Journal of Cancer Research and Clinical Oncology

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01.10.2012 | Original Paper

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

verfasst von: Guangying Du, Liang Ye, Guoying Zhang, Qiuju Dong, Kun Liu, Jingwei Tian

Erschienen in: Journal of Cancer Research and Clinical Oncology | Ausgabe 10/2012

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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.

Acres B, Gantzer M, Remy C, Futin N, Accart N, Chaloin O, Hoebeke J, Balloul JM, Paul S (2005) Fusokine interleukin-2/interleukin-18, a novel potent innate and adaptive immune stimulator with decreased toxicity. Cancer Res 65(20):9536–9546PubMedCrossRef

Blattman JN, Greenberg PD (2004) Cancer immunotherapy: a treatment for the masses. Science 305(5681):200–205PubMedCrossRef

Deng H, Nakajima K, Ma XL, Hasumi K, Akatsuka T, Wago H (2001) Anti-tumor activity of Traditional Chinese medicine, Ekki-Youketsu-Fusei-Zai, and its effects on immunocyte functions: 1. A life-prolonging effect and enhancement of NK cell function in tumor-bearing mice. J Saitama Med Sch 28(3):109–115

Dunn GP, Old LJ, Schreiber RD (2004) The immunobiology of cancer immunosurveillance and immunoediting. Immunity 21(2):137–148PubMedCrossRef

Gaffen SL, Liu KD (2004) Overview of interleukin-2 function, production and clinical applications. Cytokine 28(3):109–123PubMedCrossRef

Gu Y, Kuida K, Tsutsui H, Ku G, Hsiao K, Fleming MA, Hayashi N, Higashino K, Okamura H, Nakanishi K, Kurimoto M, Tanimoto T, Flavell RA, Sato V, Harding MW, Livingston DJ, Su MS (1997) Activation of interferon-gamma inducing factor mediated by interleukin-1beta converting enzyme. Science 275(5297):206–209PubMedCrossRef

Hoshino T, Wiltrout RH, Young HA (1999) IL-18 is a potent coinducer of IL-13 in NK and T cells: a new potential role for IL-18 in modulating the immune response. J Immunol 162(9):5070–5077PubMed

Khong HT, Restifo NP (2002) Natural selection of tumor variants in the generation of “tumor escape” phenotypes. Nat Immunol 3(11):999–1005PubMedCrossRef

Kohno K, Kataoka J, Ohtsuki T, Suemoto Y, Okamoto I, Usui M, Ikeda M, Kurimoto M (1997) IFN-gamma-inducing factor (IGIF) is a costimulatory factor on the activation of Th1 but not Th2 cells and exerts its effect independently of IL-12. J Immunol 158(4):1541–1550PubMed

Lin Y, Ya-Li Q, Wei-Cheng B, Xiao-Lan Y, Hong-Liang H, Huan-Chun C (2004) Expression of an interleukin-6—interleukin-2 fusion protein (pIL-6-IL-2) in P. pastoris. Eur Cytokine Netw 15(3):240–246PubMed

Lindsey KR, Rosenberg SA, Sherry RM (2000) Impact of the number of treatment courses on the clinical response of patients who receive high-dose bolus interleukin-2. J Clin Oncol 18(9):1954–1959PubMed

Lotze MT, Chang AE, Seipp CA, Simpson C, Vetto JT, Rosenberg SA (1986) High-dose recombinant interleukin 2 in the treatment of patients with disseminated cancer. Responses, treatment-related morbidity, and histologic findings. JAMA 256(22):3117–3124PubMedCrossRef

Lu J, Peng Y, Zheng ZJ, Pan JH, Zhang Y, Bai Y (2008) EGF-IL-18 fusion protein as a potential anti-tumor reagent by induction of immune response and apoptosis in cancer cells. Cancer Lett 260(1–2):187–197PubMedCrossRef

Margolin K, Forman SJ (2000) Immunotherapy with interleukin-2 after hematopoietic cell transplantation for hematologic malignancy. Cancer J Sci Am 6(Suppl 1):S33–S38PubMed

Micallef MJ, Tanimoto T, Kohno K, Ikeda M, Kurimoto M (1997) Interleukin 18 induces the sequential activation of natural killer cells and cytotoxic T lymphocytes to protect syngeneic mice from transplantation with Meth A sarcoma. Cancer Res 57(20):4557–4563PubMed

Morgan DA, Ruscetti FW, Gallo R (1976) Selective in vitro growth of T lymphocytes from normal human bone marrows. Science 193(4257):1007–1008PubMedCrossRef

Nakamura K, Okamura H, Wada M, Nagata K, Tamura T (1989) Endotoxin-induced serum factor that stimulates gamma interferon production. Infect Immun 57(2):590–595PubMed

Nakamura S, Otani T, Ijiri Y, Motoda R, Kurimoto M, Orita K (2000) IFN-gamma-dependent and -independent mechanisms in adverse effects caused by concomitant administration of IL-18 and IL-12. J Immunol 164(6):3330–3336PubMed

Nakamura Y, Yamada N, Ohyama H, Nakasho K, Nishizawa Y, Okamoto T, Futani H, Yoshiya S, Okamura H, Terada N (2006) Effect of interleukin-18 on metastasis of mouse osteosarcoma cells. Cancer Immunol Immunother 55(9):1151–1158PubMedCrossRef

Okamoto M, Kato S, Oizumi K, Kinoshita M, Inoue Y, Hoshino K, Akira S, McKenzie AN, Young HA, Hoshino T (2002) Interleukin 18 (IL-18) in synergy with IL-2 induces lethal lung injury in mice: a potential role for cytokines, chemokines, and natural killer cells in the pathogenesis of interstitial pneumonia. Blood 99(4):1289–1298PubMedCrossRef

Osaki T, Peron JM, Cai Q, Okamura H, Robbins PD, Kurimoto M, Lotze MT, Tahara H (1998) IFN-gamma-inducing factor/IL-18 administration mediates IFN-gamma- and IL-12-independent antitumor effects. J Immunol 160(4):1742–1749PubMed

Pan J, Zhang M, Wang J, Wang Q, Xia D, Sun W, Zhang L, Yu H, Cao X (2005) Intratumoral injection of interferon-gamma gene-modified dendritic cells elicits potent antitumor effects: effective induction of tumor-specific CD8+ CTL response. J Cancer Res Clin Oncol 131(7):468–478PubMedCrossRef

Papamichail M, Perez SA, Gritzapis AD, Baxevanis CN (2004) Natural killer lymphocytes: biology, development, and function. Cancer Immunol Immunother 53(3):176–186PubMedCrossRef

Pardoll D (2003) Does the immune system see tumors as foreign or self? Annu Rev Immunol 21:807–839PubMedCrossRef

Rosenberg SA (2004) Shedding light on immunotherapy for cancer. N Engl J Med 350(14):1461–1463PubMedCrossRef

Son YI, Dallal RM, Mailliard RB, Egawa S, Jonak ZL, Lotze MT (2001) Interleukin-18 (IL-18) synergizes with IL-2 to enhance cytotoxicity, interferon-gamma production, and expansion of natural killer cells. Cancer Res 61(3):884–888PubMed

Sone S, Ogura T (1994) Local interleukin-2 therapy for cancer, and its effector induction mechanisms. Oncology 51(2):170–176PubMedCrossRef

Ushio S, Namba M, Okura T, Hattori K, Nukada Y, Akita K, Tanabe F, Konishi K, Micallef M, Fujii M, Torigoe K, Tanimoto T, Fukuda S, Ikeda M, Okamura H, Kurimoto M (1996) Cloning of the cDNA for human IFN-gamma-inducing factor, expression in Escherichia coli, and studies on the biologic activities of the protein. J Immunol 156(11):4274–4279PubMed

Wang X, Hou M, Tan L, Sun X, Zhang Y, Li P, Zhu Y (2005) A hybrid protein of the amino-terminal fragment of urokinase and mutant plasminogen activator inhibitor-2 efficiently inhibits tumor cell invasion and metastasis. J Cancer Res Clin Oncol 131(2):129–136PubMedCrossRef

Titel: Human IL18-IL2 fusion protein as a potential antitumor reagent by enhancing NK cell cytotoxicity and IFN-γ production
verfasst von: Guangying Du
Liang Ye
Guoying Zhang
Qiuju Dong
Kun Liu
Jingwei Tian
Publikationsdatum: 01.10.2012
Verlag: Springer-Verlag
Erschienen in: Journal of Cancer Research and Clinical Oncology / Ausgabe 10/2012
Print ISSN: 0171-5216
Elektronische ISSN: 1432-1335
DOI: https://doi.org/10.1007/s00432-012-1248-5

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

Abstract

Purpose

Methods

Results

Conclusions

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Springer Medizin

Abstract

Purpose

Methods

Results

Conclusions

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