Ex vivo generation of highly purified and activated natural killer cells from human peripheral blood

Satoru Saito, Yui Harada, Yosuke Morodomi, Mitsuho Onimaru, Kumi Yoshida, Ryoichi Kyuragi, Hisahiro Matsubara, Yoshikazu Yonemitsu

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Adoptive immunotherapy using natural killer (NK) cells has been a promising treatment for intractable malignancies; however, there remain a number of difficulties with respect to the shortage and limited anticancer potency of the effector cells. We here established a simple feeder-free method to generate purified (>90%) and highly activated NK cells from human peripheral blood-derived mononuclear cells (PBMCs). Among the several parameters, we found that CD3 depletion, high-dose interleukin (IL)-2, and use of a specific culture medium were sufficient to obtain highly purified, expanded (∼200-fold) and activated CD3-/CD56+ NK cells from PBMCs, which we designated zenithal-NK (Z-NK) cells. Almost all Z-NK cells expressed the lymphocyte-activated marker CD69 and showed dramatically high expression of activation receptors (i.e., NKG2D), interferon-γ, perforin, and granzyme B. Importantly, only 2 hours of reaction at an effector/target ratio of 1:1 was sufficient to kill almost all K562 cells, and the antitumor activity was also replicated in tumor-bearing mice in vivo. Cytolysis was specific for various tumor cells, but not for normal cells, irrespective of MHC class I expression. These findings strongly indicate that Z-NK cells are purified, expanded, and near-fully activated human NK cells and warrant further investigation in a clinical setting.

Original languageEnglish
Pages (from-to)241-252
Number of pages12
JournalHuman Gene Therapy Methods
Volume24
Issue number4
DOIs
Publication statusPublished - Aug 1 2013

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Natural Killer Cells
NK Cell Lectin-Like Receptor Subfamily K
Adoptive Immunotherapy
Neoplasms
K562 Cells
Interferons
Interleukin-2
Culture Media
Lymphocytes

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Applied Microbiology and Biotechnology
  • Genetics
  • Pharmacology
  • Genetics(clinical)

Cite this

Ex vivo generation of highly purified and activated natural killer cells from human peripheral blood. / Saito, Satoru; Harada, Yui; Morodomi, Yosuke; Onimaru, Mitsuho; Yoshida, Kumi; Kyuragi, Ryoichi; Matsubara, Hisahiro; Yonemitsu, Yoshikazu.

In: Human Gene Therapy Methods, Vol. 24, No. 4, 01.08.2013, p. 241-252.

Research output: Contribution to journalArticle

Saito, Satoru ; Harada, Yui ; Morodomi, Yosuke ; Onimaru, Mitsuho ; Yoshida, Kumi ; Kyuragi, Ryoichi ; Matsubara, Hisahiro ; Yonemitsu, Yoshikazu. / Ex vivo generation of highly purified and activated natural killer cells from human peripheral blood. In: Human Gene Therapy Methods. 2013 ; Vol. 24, No. 4. pp. 241-252.
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