Abstract
NK cells hold promise for protecting hosts from cancer and pathogen infection through direct killing and expressing immune-regulatory cytokines. In our study, a genetically modified K562 cell line with surface expression of 4-1BBL and MICA was constructed to expand functional NK cells in vitro for further adoptive immunotherapy against cancer. After a long-term up to 21 day co-culture with newly isolated peripheral blood mononuclear cells (PBMCs) in the presence of soluble IL-21 (sIL-21), notable increase in proportion of expanded NK cells was observed, especially the CD56(bright)CD16(+) subset. Apparent up-regulation of activating receptors CD38, CD69 and NKG2D was detected on expanded NK cells, so did inhibitory receptor CD94; the cytotoxicity of expanded NK cells against target tumor cells exceeded that of NK cells within fresh PBMCs. The intracellular staining showed expanded NK cells produced immune-regulatory IFN-γ. Taken together, we expanded NK cells with significant up-regulation of activating NKG2D and moderate enhancement of cytotoxicity, with IFN-γ producing ability and a more heterogeneous population of NK cells. These findings provide a novel perspective on expanding NK cells in vitro for further biology study and adoptive immunotherapy of NK cells against cancer.
Keywords:
4-1BBL; IL-21; K562 cells; MICA; NK cells.
Copyright © 2014 Elsevier Inc. All rights reserved.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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4-1BB Ligand / biosynthesis*
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4-1BB Ligand / genetics
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ADP-ribosyl Cyclase 1 / biosynthesis
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Antigens, CD / biosynthesis
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Antigens, Differentiation, T-Lymphocyte / biosynthesis
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CD56 Antigen / biosynthesis
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Cell Line, Tumor
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Coculture Techniques
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GPI-Linked Proteins / biosynthesis
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HeLa Cells
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Hep G2 Cells
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Histocompatibility Antigens Class I / biosynthesis*
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Histocompatibility Antigens Class I / genetics
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Humans
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Immunotherapy, Adoptive
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Interferon-gamma / biosynthesis
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Interferon-gamma / immunology
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Interleukins / biosynthesis*
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Interleukins / genetics
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Interleukins / pharmacology
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Killer Cells, Natural / immunology*
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Lectins, C-Type / biosynthesis
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Lymphocyte Activation / immunology
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Membrane Glycoproteins / biosynthesis
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NK Cell Lectin-Like Receptor Subfamily D / biosynthesis
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NK Cell Lectin-Like Receptor Subfamily K / biosynthesis
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Neoplasms / immunology*
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Neoplasms / therapy
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Receptors, IgG / biosynthesis
Substances
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4-1BB Ligand
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Antigens, CD
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Antigens, Differentiation, T-Lymphocyte
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CD56 Antigen
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CD69 antigen
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FCGR3B protein, human
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GPI-Linked Proteins
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Histocompatibility Antigens Class I
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Interleukins
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KLRD1 protein, human
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KLRK1 protein, human
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Lectins, C-Type
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MHC class I-related chain A
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Membrane Glycoproteins
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NCAM1 protein, human
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NK Cell Lectin-Like Receptor Subfamily D
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NK Cell Lectin-Like Receptor Subfamily K
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Receptors, IgG
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Interferon-gamma
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CD38 protein, human
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ADP-ribosyl Cyclase 1
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interleukin-21