title: Phase I clinical trial of autologous NK cell therapy using novel expansion method in patients with advanced digestive cancer.
abstract: BACKGROUND
  NK cells can destroy tumor cells without prior sensitization or immunization. Tumors often lose expression of MHC molecules and/or antigens. However, NK cells can lyse tumor cells in a non-MHC-restricted manner and independent of the expression of tumor-associated antigens. NK cells are therefore considered ideal for adoptive cancer immunotherapy; however the difficulty of obtaining large numbers of fully functional NK cells that are safe to administer deters its clinical use. This phase I clinical trial seeks to address this obstacle by first developing a novel system that expands large numbers of highly activated clinical grade NK cells, and second, determining if these cells are safe in a mono-treatment so they can be combined with other reagents in the next round of clinical trials.
  
  
  METHODS
  Patients with unresectable, locally advanced and/or metastatic digestive cancer who did not succeed with standard therapy were enrolled. NK cells were expanded ex vivo by stimulating PBMCs with OK432, IL-2, and modified FN-CH296 induced T cells. Patients were administered autologous natural killer cell three times weekly via intravenous infusions in a dose-escalating manner (dose 0.5 × 10(9), 1.0 × 10(9), 2.0 × 10(9) cells/injection, three patients/one cohort).
  
  
  RESULTS
  Total cell population had a median expansion of 586-fold (range 95-1102), with a significantly pure (90.96 %) NK cell population. Consequently, NK cells were expanded to approximately 4720-fold (range 1372-14,116) with cells being highly lytic in vitro and strongly expressing functional markers such as NKG2D and CD16. This NK cell therapy was very well tolerated with no severe adverse events. Although no clinical responses were observed, cytotoxicity of peripheral blood was elevated approximately twofolds up to 4 weeks post the last transfer.
  
  
  CONCLUSIONS
  We successfully generated large numbers of activated NK cells from small quantities of blood without prior purification of the cells. We also determined that the expanded cells were safe to administer in a monotherapy and are suitable for the next round of clinical trials where their efficacy will be tested combined with other reagents.
  
  
  BACKGROUND
  UMIN UMIN000007527.
authors: Department of Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Japan. jrsaka@koto.kpu-m.ac.jp.|Iseikai Hyakumanben Clinic, Kyoto, Japan. jrsaka@koto.kpu-m.ac.jp.
authors: Naoyuki
authors: N
authors: Sakamoto
authors: Department of Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Japan. iskw-t@koto.kpu-m.ac.jp.|Department of Cancer ImmunoCell Regulation, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, 602-8566, Japan. iskw-t@koto.kpu-m.ac.jp.
authors: Takeshi
authors: T
authors: Ishikawa
authors: Department of Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Japan. s-kokura@koto.kpu-m.ac.jp.|Center for Education Research and Development, Kyoto Gakuen University, Kyoto, Japan. s-kokura@koto.kpu-m.ac.jp.
authors: Satoshi
authors: S
authors: Kokura
authors: Department of Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Japan. t-oka@koto.kpu-m.ac.jp.|Department of Cancer ImmunoCell Regulation, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, 602-8566, Japan. t-oka@koto.kpu-m.ac.jp.
authors: Tetsuya
authors: T
authors: Okayama
authors: Department of Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Japan. k-oka@koto.kpu-m.ac.jp.
authors: Kaname
authors: K
authors: Oka
authors: CDM Center, Takara Bio Inc, Otsu, Japan. idenom@takara-bio.co.jp.
authors: Mitsuko
authors: M
authors: Ideno
authors: CDM Center, Takara Bio Inc, Otsu, Japan. sakaif@takara-bio.co.jp.
authors: Fumiyo
authors: F
authors: Sakai
authors: CDM Center, Takara Bio Inc, Otsu, Japan. katoaz@takara-bio.co.jp.
authors: Akiko
authors: A
authors: Kato
authors: CDM Center, Takara Bio Inc, Otsu, Japan. tanabemz@takara-bio.co.jp.
authors: Masashige
authors: M
authors: Tanabe
authors: CDM Center, Takara Bio Inc, Otsu, Japan. enokit@takara-bio.co.jp.
authors: Tatsuji
authors: T
authors: Enoki
authors: CDM Center, Takara Bio Inc, Otsu, Japan. minenoj@takara-bio.co.jp.
authors: Junichi
authors: J
authors: Mineno
authors: Department of Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Japan. ynaito@koto.kpu-m.ac.jp.
authors: Yuji
authors: Y
authors: Naito
authors: Department of Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Japan. yitoh@koto.kpu-m.ac.jp.
authors: Yoshito
authors: Y
authors: Itoh
authors: Department of Cancer ImmunoCell Regulation, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, 602-8566, Japan. toshi@koto.kpu-m.ac.jp.
authors: Toshikazu
authors: T
authors: Yoshikawa
mesh_terms: D000368:Aged; D005260:Female; D005770:Gastrointestinal Neoplasms / Q000628:therapy*; D006801:Humans; D016130:Immunophenotyping; D007167:Immunotherapy; D007694:Killer Cells, Natural / Q000166:cytology* / Q000276:immunology; D008297:Male; D008875:Middle Aged