An IL-12/Shh-C domain fusion protein-based IL-12 autocrine loop for sustained natural killer cell activation

Zhu,Lining; Zhao,Zhihui; Wei,Yanzhang; Marcotte ,William; Wagner,Thomas  E.; Yu,Xianzhong

doi:10.3892/ijo.2012.1466

An IL-12/Shh-C domain fusion protein-based IL-12 autocrine loop for sustained natural killer cell activation

Authors:
- Lining Zhu
- Zhihui Zhao
- Yanzhang Wei
- William Marcotte
- Thomas E. Wagner
- Xianzhong Yu
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Affiliations: Department of Biological Sciences, Clemson University, Clemson, SC 29634, USA, Department of Animal Genetics and Breeding, Jilin University, Changchun 130062, P.R. China, Department of Genetics and Biochemistry, Clemson University, Clemson, SC 29634, USA, Orbis Health Solutions, Greenville, SC 29607, USA
Published online on: May 8, 2012 https://doi.org/10.3892/ijo.2012.1466
Pages: 661-669

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Abstract

The dependency of activated natural killer (NK) cells on the continuous support of exogenous interleukin (IL)-2 for their in vivo survival, tumor localization and consequently, their antitumor effect, is a major obstacle for NK cell-mediated tumor therapy. In the present study, a fusion gene between IL-12 and mouse sonic hedgehog C-terminal domain (Shh-C) was constructed. The fusion protein was autocatalytically processed to form cholesterol-modified IL-12 molecules and an autocrine loop of IL-12 was established for the sustained activation of NK cells. The transduced NK cells matured more rapidly in vitro with the enhanced expression of granule-related proteins. NKIL-12/Shh-C cells reached the same proliferation rate as NK cells transduced with enhanced green fluorescent protein (EGFP)/Shh-C (NKEGFP/Shh-C) with <10-fold IL-2 supplementation, suggesting that the fusion protein reduced the dependency of NK cells on IL-2. The amount of interferon‑γ (IFN-γ) in the supernatants of NKIL-12/Shh-C cells 5 and 7 days after transduction was significantly higher than that in the supernatants of NKIL-12 cells. Immunofluorescent staining of lung tissues from B16-bearing mice which had received an intravenous injection of lentivirus-transduced NK cells without exogenous IL-2 confirmed that donor NK cells successfully infiltrated into the lung tissues. The survival time of the mice which had received NKIL-12/Shh-C cells was significantly prolonged compared to the mice which had received NKEGFP/Shh-C cells.

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