Skip to main content


Log in

Pilot study to determine the safety and feasibility of deceased donor liver natural killer cell infusion to liver transplant recipients with hepatocellular carcinoma

  • Original Article
  • Published:
Cancer Immunology, Immunotherapy Aims and scope Submit manuscript


Liver transplantation (LT) is a viable treatment option for cirrhosis patients with hepatocellular carcinoma (HCC). However, recurrence is the rate-limiting factor of long-term survival. To prevent this, we conducted the phase I study of the adoptive transfer of deceased donor liver-derived natural killer (NK) cells. Liver NK cells were extracted from donor liver graft perfusate and were stimulated in vitro with IL-2. The patient received an intravenous infusion of NK cells 3–5 days after LT. Eighteen LT recipients were treated. There were no severe cell infusion-related adverse events or acute rejection episodes. One patient withdrew from the study because the pathological observation revealed sarcoma instead of HCC. All patients who received this immunotherapy completed the follow-up for at least 2 years without evidence of HCC recurrence (median follow-up, 96 months [range, 17–121 months]). Considering that 9 (52.9%) of the 17 patients pathologically exceeded the Milan criteria, liver NK cell infusion is likely to be useful for preventing HCC recurrence after LT. This is the first-in-human immunotherapy study using deceased donor liver-derived NK cells to prevent HCC recurrence after LT. This treatment was well tolerated and resulted in no HCC recurrence after LT.

Clinical trial registration; NCT01147380; registration date: June 17, 2010.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others



Cytokine-induced killer


Common Terminology Criteria for Adverse Events


Deceased donor liver transplantation


Flow cytometry


Graft-versus-host disease


Hepatocellular carcinoma


Human leukocyte antigen


Killer cell immunoglobulin-like receptor


Living donor liver transplantation


Liver mononuclear cells


Liver transplantation


The Model for End-Stage Liver Disease


Natural killer


Peripheral blood mononuclear cells


Tumor necrosis factor-related apoptosis-inducing ligand


The United Network for Organ Sharing


  1. Mazzaferro V, Regalia E, Doci R, Andreola S, Pulvirenti A, Bozzetti F et al (1996) Liver transplantation for the treatment of small hepatocellular carcinomas in patients with cirrhosis. N Engl J Med 334(11):693–699.

    Article  CAS  PubMed  Google Scholar 

  2. Ioannou GN, Perkins JD, Carithers RL Jr (2008) Liver transplantation for hepatocellular carcinoma: impact of the MELD allocation system and predictors of survival. Gastroenterology 134(5):1342–1351.

    Article  PubMed  Google Scholar 

  3. Saab S, Yeganeh M, Nguyen K, Durazo F, Han S, Yersiz H et al (2009) Recurrence of hepatocellular carcinoma and hepatitis B reinfection in hepatitis B surface antigen-positive patients after liver transplantation. Liver Transpl 15(11):1525–1534.

    Article  PubMed  Google Scholar 

  4. Levi DM, Tzakis AG, Martin P, Nishida S, Island E, Moon J et al (2010) Liver transplantation for hepatocellular carcinoma in the model for end-stage liver disease era. J Am Coll Surg 210(5):727–734.

    Article  PubMed  Google Scholar 

  5. Horn M, Phebus C, Blatt J (1990) Cancer chemotherapy after solid organ transplantation. Cancer 66(7):1468–1471.;2-b

    Article  CAS  PubMed  Google Scholar 

  6. Vivarelli M, Cucchetti A, La Barba G, Ravaioli M, Del Gaudio M, Lauro A et al (2008) Liver transplantation for hepatocellular carcinoma under calcineurin inhibitors: reassessment of risk factors for tumor recurrence. Ann Surg 248(5):857–862.

    Article  PubMed  Google Scholar 

  7. Yokoyama I, Carr B, Saitsu H, Iwatsuki S, Starzl TE (1991) Accelerated growth rates of recurrent hepatocellular carcinoma after liver transplantation. Cancer 68(10):2095–2100.;2-y

    Article  CAS  PubMed  Google Scholar 

  8. Soderdahl G, Backman L, Isoniemi H, Cahlin C, Hockerstedt K, Broome U et al (2006) A prospective, randomized, multi-centre trial of systemic adjuvant chemotherapy versus no additional treatment in liver transplantation for hepatocellular carcinoma. Transpl Int 19(4):288–294.

    Article  CAS  PubMed  Google Scholar 

  9. Zimmerman, M. A., Ghobrial, R. M., Tong, M. J., Hiatt, J. R., Cameron, A. M., Hong, J., et al. (2008). Recurrence of hepatocellular carcinoma following liver transplantation: a review of preoperative and postoperative prognostic indicators. Arch Surg 143(2):182–188; discussion 188.

  10. Ljunggren HG, Karre K (1990) In search of the “missing self”: MHC molecules and NK cell recognition. Immunol Today 11(7):237–244.

    Article  CAS  PubMed  Google Scholar 

  11. Karre K (2002) NK cells, MHC class I molecules and the missing self. Scand J Immunol 55(3):221–228.

    Article  CAS  PubMed  Google Scholar 

  12. Balch CM, Tilden AB, Dougherty PA, Cloud GA, Abo T (1984) Heterogeneity of natural killer lymphocyte abnormalities in colon cancer patients. Surgery 95(1):63–70

    CAS  PubMed  Google Scholar 

  13. Cai L, Zhang Z, Zhou L, Wang H, Fu J, Zhang S et al (2008) Functional impairment in circulating and intrahepatic NK cells and relative mechanism in hepatocellular carcinoma patients. Clin Immunol 129(3):428–437.

    Article  CAS  PubMed  Google Scholar 

  14. Ishiyama K, Ohdan H, Ohira M, Mitsuta H, Arihiro K, Asahara T (2006) Difference in cytotoxicity against hepatocellular carcinoma between liver and periphery natural killer cells in humans. Hepatology 43(2):362–372.

    Article  CAS  PubMed  Google Scholar 

  15. Harada N, Shimada M, Okano S, Suehiro T, Soejima Y, Tomita Y et al (2004) IL-12 gene therapy is an effective therapeutic strategy for hepatocellular carcinoma in immunosuppressed mice. J Immunol 173(11):6635–6644.

    Article  CAS  PubMed  Google Scholar 

  16. Hirata M, Kita Y, Saito S, Nishimura M, Ito M, Mizuta K et al (1998) Increase in natural killer cell activity following living-related liver transplantation. Transpl Int 11(Suppl 1):S185-188.

    Article  PubMed  Google Scholar 

  17. Miller JS, Soignier Y, Panoskaltsis-Mortari A, McNearney SA, Yun GH, Fautsch SK et al (2005) Successful adoptive transfer and in vivo expansion of human haploidentical NK cells in patients with cancer. Blood 105(8):3051–3057.

    Article  CAS  PubMed  Google Scholar 

  18. Ohira M, Ishiyama K, Tanaka Y, Doskali M, Igarashi Y, Tashiro H et al (2009) Adoptive immunotherapy with liver allograft-derived lymphocytes induces anti-HCV activity after liver transplantation in humans and humanized mice. J Clin Invest 119(11):3226–3235.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Ohira M, Ohdan H, Mitsuta H, Ishiyama K, Tanaka Y, Igarashi Y et al (2006) Adoptive transfer of TRAIL-expressing natural killer cells prevents recurrence of hepatocellular carcinoma after partial hepatectomy. Transplantation 82(12):1712–1719.

    Article  CAS  PubMed  Google Scholar 

  20. Xu L, Wang J, Kim Y, Shuang ZY, Zhang YJ, Lao XM et al (2016) A randomized controlled trial on patients with or without adjuvant autologous cytokine-induced killer cells after curative resection for hepatocellular carcinoma. Oncoimmunology 5(3):e1083671.

    Article  PubMed  Google Scholar 

  21. Li X, Dai D, Song X, Liu J, Zhu L, Xu W (2014) A meta-analysis of cytokine-induced killer cells therapy in combination with minimally invasive treatment for hepatocellular carcinoma. Clin Res Hepatol Gastroenterol 38(5):583–591.

    Article  CAS  PubMed  Google Scholar 

  22. Lee JH, Lee JH, Lim YS, Yeon JE, Song TJ, Yu SJ et al (2015) Adjuvant immunotherapy with autologous cytokine-induced killer cells for hepatocellular carcinoma. Gastroenterology 148(7):1383-1391 e1386.

    Article  CAS  PubMed  Google Scholar 

  23. Rubnitz JE, Inaba H, Ribeiro RC, Pounds S, Rooney B, Bell T et al (2010) NKAML: a pilot study to determine the safety and feasibility of haploidentical natural killer cell transplantation in childhood acute myeloid leukemia. J Clin Oncol 28(6):955–959.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Brand JM, Meller B, Von Hof K, Luhm J, Bahre M, Kirchner H et al (2004) Kinetics and organ distribution of allogeneic natural killer lymphocytes transfused into patients suffering from renal cell carcinoma. Stem Cells Dev 13(3):307–314.

    Article  CAS  PubMed  Google Scholar 

  25. Tanimine N, Tanaka Y, Kobayashi T, Tashiro H, Miki D, Imamura M et al (2014) Quantitative effect of natural killer-cell licensing on hepatocellular carcinoma recurrence after curative hepatectomy. Cancer Immunol Res 2(12):1142–1147.

    Article  CAS  PubMed  Google Scholar 

  26. Ohira M, Nishida S, Matsuura T, Muraoka I, Tryphonopoulos P, Fan J et al (2013) Comparative analysis of T-cell depletion method for clinical immunotherapy-anti-hepatitis c effects of natural killer cells via interferon-gamma production. Transplant Proc 45(5):2045–2050.

    Article  CAS  PubMed  Google Scholar 

  27. Ohira M, Nishida S, Tryphonopoulos P, Tekin A, Selvaggi G, Moon J et al (2012) Clinical-scale isolation of interleukin-2-stimulated liver natural killer cells for treatment of liver transplantation with hepatocellular carcinoma. Cell Transplant 21(7):1397–1406.

    Article  PubMed  Google Scholar 

  28. Kumar A, Le DT (2016) Hepatocellular Carcinoma Regression After Cessation of Immunosuppressive Therapy. J Clin Oncol 34(10):e90-92.

    Article  CAS  PubMed  Google Scholar 

  29. Shimizu S, Tanaka Y, Tazawa H, Verma S, Onoe T, Ishiyama K et al (2016) Fc-gamma receptor polymorphisms predispose patients to infectious complications after liver transplantation. Am J Transplant 16(2):625–633.

    Article  CAS  PubMed  Google Scholar 

  30. Frohn C, Doehn C, Durek C, Bohle A, Schlenke P, Jocham D et al (2000) Feasibility of the adoptive transfusion of allogenic human leukocyte antigen-matched natural killer cells in patients with renal cell carcinoma. J Immunother 23(4):499–504.

    Article  CAS  PubMed  Google Scholar 

  31. Schulze A, Schirutschke H, Oelschlagel U, Schmitz M, Fussel M, Wassmuth R et al (2008) Altered phenotype of natural killer cell subsets after haploidentical stem cell transplantation. Exp Hematol 36(4):378–389.

    Article  CAS  PubMed  Google Scholar 

  32. Passweg JR, Tichelli A, Meyer-Monard S, Heim D, Stern M, Kuhne T et al (2004) Purified donor NK-lymphocyte infusion to consolidate engraftment after haploidentical stem cell transplantation. Leukemia 18(11):1835–1838.

    Article  CAS  PubMed  Google Scholar 

  33. Sakamoto N, Ishikawa T, Kokura S, Okayama T, Oka K, Ideno M et al (2015) Phase I clinical trial of autologous NK cell therapy using novel expansion method in patients with advanced digestive cancer. J Transl Med 13:277.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Doskali M, Tanaka Y, Ohira M, Ishiyama K, Tashiro H, Chayama K et al (2011) Possibility of adoptive immunotherapy with peripheral blood-derived CD3(-)CD56+ and CD3+CD56+ cells for inducing antihepatocellular carcinoma and antihepatitis C virus activity. J Immunother 34(2):129–138.

    Article  CAS  PubMed  Google Scholar 

  35. Pan G, O’Rourke K, Chinnaiyan AM, Gentz R, Ebner R, Ni J et al (1997) The receptor for the cytotoxic ligand TRAIL. Science 276(5309):111–113.

    Article  CAS  PubMed  Google Scholar 

  36. Sheridan JP, Marsters SA, Pitti RM, Gurney A, Skubatch M, Baldwin D et al (1997) Control of TRAIL-induced apoptosis by a family of signaling and decoy receptors. Science 277(5327):818–821.

    Article  CAS  PubMed  Google Scholar 

  37. Mikulak J, Bruni E, Oriolo F, Di Vito C, Mavilio D (2019) Hepatic Natural Killer Cells: Organ-Specific Sentinels of Liver Immune Homeostasis and Physiopathology. Front Immunol 10:946.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references


We would like to thank Oliver Umland for supporting flow cytometry analysis, Drs. Ryosuke Misawa, Taizo Hibi, Koichiro Uchida, Takehiko Dohi, Izumi Carpenter, Ji Fan, David Grant, Panagiotis Tryphonopoulos, Bonnie Blomberg, and Gary Kleiner for study support, and Editage ( for English language editing.


This study was supported by the research funding the Grant No. 1BG-08 from the Florida Department of Health and the Bankhead-Coley Cancer Research Program (Seigo Nishida), AMED under Grant Number JP21fk0210051 (Hideki Ohdan), and JSPS KAKENHI Grant Number JP20K09104 (Masahiro Ohira).

Author information

Authors and Affiliations


Corresponding authors

Correspondence to Andreas G. Tzakis or Hideki Ohdan.

Ethics declarations

Conflict of interest

The authors of this manuscript have no conflict of interest to disclose as described by the Cancer Immunology, Immunotherapy.

Ethical approval

This phase I clinical trial was approved by the Institutional Review Board of the University of Miami Miller School of Medicine (IRB#20100344) and the Food and Drug Administration, and was registered with (NCT01147380). The trial was designed and conducted according to the Declaration of Helsinki.

Informed consent

All patients provided written informed consent before enrolling in the study.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (PDF 221 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ohira, M., Hotta, R., Tanaka, Y. et al. Pilot study to determine the safety and feasibility of deceased donor liver natural killer cell infusion to liver transplant recipients with hepatocellular carcinoma. Cancer Immunol Immunother 71, 589–599 (2022).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: