title: Clinical-scale isolation of interleukin-2-stimulated liver natural killer cells for treatment of liver transplantation with hepatocellular carcinoma.
abstract: 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 cell-mediated 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 ± 10(6) 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.
authors.affiliation: Department of Surgery, Division of Liver and Gastrointestinal Transplantation, University of Miami Miller School of Medicine, 1801 NW 9th Avenue, Miami, FL 33136, USA.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Ohira
authors.forename: Seigo
authors.initials: S
authors.lastname: Nishida
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Tryphonopoulos
authors.forename: Akin
authors.initials: A
authors.lastname: Tekin
authors.forename: Gennaro
authors.initials: G
authors.lastname: Selvaggi
authors.forename: Jang
authors.initials: J
authors.lastname: Moon
authors.forename: David
authors.initials: D
authors.lastname: Levi
authors.forename: Camillo
authors.initials: C
authors.lastname: Ricordi
authors.forename: Kohei
authors.initials: K
authors.lastname: Ishiyama
authors.forename: Yuka
authors.initials: Y
authors.lastname: Tanaka
authors.forename: Hideki
authors.initials: H
authors.lastname: Ohdan
authors.forename: Andreas G
authors.initials: AG
authors.lastname: Tzakis
mesh_terms: D000328:Adult; D000368:Aged; D015703:Antigens, CD / Q000378:metabolism; D000945:Antigens, Differentiation, T-Lymphocyte / Q000378:metabolism; D017209:Apoptosis; D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D016207:Cytokines / Q000378:metabolism; D005260:Female; D006084:Graft Rejection; D006801:Humans; D007167:Immunotherapy; D007376:Interleukin-2 / Q000494:pharmacology*; D020014:K562 Cells; D007694:Killer Cells, Natural / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D037181:Lectins, C-Type / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000166:cytology; D008099:Liver / Q000166:cytology / Q000378:metabolism; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D016031:Liver Transplantation; D008297:Male; D008875:Middle Aged; D055627:Natural Cytotoxicity Triggering Receptor 1 / Q000378:metabolism; D055629:Natural Cytotoxicity Triggering Receptor 2 / Q000378:metabolism; D055502:Secondary Prevention; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000378:metabolism; D014019:Tissue Donors; D015854:Up-Regulation / Q000187:drug effects; D055815:Young Adult; D000099192:CD69 Antigens
chemical_list: D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D016207:Cytokines; D007376:Interleukin-2; D037181:Lectins, C-Type; D055627:Natural Cytotoxicity Triggering Receptor 1; D055629:Natural Cytotoxicity Triggering Receptor 2; D053221:TNF-Related Apoptosis-Inducing Ligand; D000099192:CD69 Antigens