Clinical-scale isolation of interleukin-2-stimulated liver natural killer cells for treatment of liver transplantation with hepatocellular carcinoma

Masahiro Ohira, Seigo Nishida, Panagiotis Tryphonopoulos, Akin Tekin, Gennaro Selvaggi, Jang Moon, David Levi, Camillo Ricordi, Kohei Ishiyama, Yuka Tanaka, Hideki Ohdan, Andreas G. Tzakis

Research output: Contribution to journalArticle

23 Scopus citations


Tumor recurrence is the main limitation of liver transplantation (LT) in patients with hepatocellular carcinoma (HCC) and can be promoted by immunosuppressants. However, there is no prevention or treatment for HCC recurrence after LT. Here we describe a clinical-scale method for an adoptive immunotherapy approach that uses natural killer (NK) cells derived from deceased donor liver graft perfusate to prevent tumor recurrence after LT. Liver mononuclear cells (LMNCs) that were extracted from deceased donor liver graft perfusate contained a high percentage of NK cells (45.0 ± 4.0%) compared with peripheral blood mononuclear cells (PBMCs) (21.8 ± 5.2%) from the same donor. The CD69 activation marker and the natural cytotoxicity receptors, NKp44 and NKp46, were expressed at high levels in freshly isolated liver NK cells. Furthermore, interleukin-2 (IL-2)-stimulated NK cells showed greater upregulation of activation markers and the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), which is critical for NK cellmediated antitumor cell death and increased production of interferon. Moreover, IL-2 stimulation induced LMNCs to exhibit a strong cytotoxicity against NK-susceptible K562 target cells compared with PBMCs (p < 0.01). Finally, we also showed that the final product contained a very low T-cell contamination (0.02 ± 106 cells/kg-1), which reduces the risk of graft-versus-host disease (GVHD). Collectively, our results suggest that the adoptive transfer of IL-2-stimulated NK cells from deceased donor liver graft perfusate could be a promising treatment for LT patients with HCC.

Original languageEnglish (US)
Pages (from-to)1397-1406
Number of pages10
JournalCell Transplantation
Issue number7
StatePublished - Nov 12 2012



  • Current good manufacturing practice (cGMP)
  • Hepatocellular carcinoma
  • Immunotherapy
  • Innate immunity
  • Natural killer cell

ASJC Scopus subject areas

  • Cell Biology
  • Transplantation
  • Biomedical Engineering

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