Abstract
Natural killer (NK) cells are important mediators of anti-tumor immunity and are active against several hematologic malignancies, including multiple myeloma (MM). Umbilical cord blood (CB) is a promising source of allogeneic NK cells but large scale ex vivo expansion is required for generation of clinically relevant CB-derived NK (CB-NK) cell doses. Here we describe a novel strategy for expanding NK cells from cryopreserved CB units using artificial antigen presenting feeder cells (aAPC) in a gas permeable culture system. After 14 days, mean fold expansion of CB-NK cells was 1848-fold from fresh and 2389-fold from cryopreserved CB with >95% purity for NK cells (CD56(+)/CD3(-)) and less than 1% CD3(+) cells. Though surface expression of some cytotoxicity receptors was decreased, aAPC-expanded CB-NK cells exhibited a phenotype similar to CB-NK cells expanded with IL-2 alone with respect to various inhibitory receptors, NKG2C and CD94 and maintained strong expression of transcription factors Eomesodermin and T-bet. Furthermore, CB-NK cells formed functional immune synapses with and demonstrated cytotoxicity against various MM targets. Finally, aAPC-expanded CB-NK cells showed significant in vivo activity against MM in a xenogenic mouse model. Our findings introduce a clinically applicable strategy for the generation of highly functional CB-NK cells which can be used to eradicate MM.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antigen-Presenting Cells / immunology*
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CD3 Complex / immunology
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CD3 Complex / metabolism
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CD56 Antigen / immunology
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CD56 Antigen / metabolism
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Cell Culture Techniques
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Cell Line, Tumor
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Cell Proliferation*
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Cells, Cultured
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Coculture Techniques
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Cytotoxicity, Immunologic / drug effects
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Cytotoxicity, Immunologic / immunology
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Fetal Blood / cytology
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Fetal Blood / immunology*
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Fetal Blood / metabolism
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Humans
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Interleukin Receptor Common gamma Subunit / deficiency
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Interleukin Receptor Common gamma Subunit / genetics
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Interleukin Receptor Common gamma Subunit / immunology
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Interleukin-2 / immunology
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Interleukin-2 / pharmacology
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K562 Cells
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Killer Cells, Natural / immunology*
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Killer Cells, Natural / metabolism
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Mice
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Mice, Inbred NOD
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Mice, Knockout
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Mice, SCID
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Microscopy, Confocal
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Multiple Myeloma / immunology*
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Multiple Myeloma / pathology
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Multiple Myeloma / therapy
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NK Cell Lectin-Like Receptor Subfamily C / immunology
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NK Cell Lectin-Like Receptor Subfamily C / metabolism
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NK Cell Lectin-Like Receptor Subfamily D / immunology
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NK Cell Lectin-Like Receptor Subfamily D / metabolism
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Xenograft Model Antitumor Assays / methods
Substances
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CD3 Complex
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CD56 Antigen
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Interleukin Receptor Common gamma Subunit
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Interleukin-2
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KLRC2 protein, human
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NK Cell Lectin-Like Receptor Subfamily C
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NK Cell Lectin-Like Receptor Subfamily D