RRC ID 64115
Author Saito S, Harada Y, Morodomi Y, Onimaru M, Yoshida K, Kyuragi R, Matsubara H, Yonemitsu Y.
Title Ex vivo generation of highly purified and activated natural killer cells from human peripheral blood.
Journal Hum Gene Ther Methods
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.
Volume 24(4)
Pages 241-52
Published 2013-8-1
DOI 10.1089/hgtb.2012.183
PMID 23885718
PMC PMC3753730
MeSH Animals Antigens, CD / genetics Antigens, CD / metabolism Antigens, Differentiation, T-Lymphocyte / genetics Antigens, Differentiation, T-Lymphocyte / metabolism Female Flow Cytometry / methods* Humans Interferon-gamma / genetics Interferon-gamma / metabolism K562 Cells Killer Cells, Lymphokine-Activated / cytology* Killer Cells, Lymphokine-Activated / immunology Killer Cells, Lymphokine-Activated / metabolism Lectins, C-Type / genetics Lectins, C-Type / metabolism Mice Mice, Inbred NOD Xenograft Model Antitumor Assays
IF 2.446
Resource
Human and Animal Cells MRC-5