Expansion of NK cells by engineered K562 cells co-expressing 4-1BBL and mMICA, combined with soluble IL-21.

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.

• Authors: Bo Jiang, Xuan Wu, Xi-Ning Li, Xi Yang, Yulai Zhou, Haowei Yan, An-Hui Wei, Weiqun Yan
• Journal: Cellular immunology (Cell Immunol) Vol. 290 Issue 1 Pg. 10-20 (Jul 2014) ISSN: 1090-2163 [Electronic] Netherlands
• PMID: 24859012 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2014 Elsevier Inc. All rights reserved.

Chemical References

• 4-1BB Ligand
• Antigens, CD
• Antigens, Differentiation, T-Lymphocyte
• CD56 Antigen
• CD69 antigen
• FCGR3B protein, human
• GPI-Linked Proteins
• Histocompatibility Antigens Class I
• Interleukins
• KLRD1 protein, human
• KLRK1 protein, human
• Lectins, C-Type
• MHC class I-related chain A
• Membrane Glycoproteins
• NCAM1 protein, human
• NK Cell Lectin-Like Receptor Subfamily D
• NK Cell Lectin-Like Receptor Subfamily K
• Receptors, IgG
• Interferon-gamma
• CD38 protein, human
• ADP-ribosyl Cyclase 1
• interleukin-21

Topics

• 4-1BB Ligand (biosynthesis, genetics)
• ADP-ribosyl Cyclase 1 (biosynthesis)
• Antigens, CD (biosynthesis)
• Antigens, Differentiation, T-Lymphocyte (biosynthesis)
• CD56 Antigen (biosynthesis)
• Cell Line, Tumor
• Coculture Techniques
• GPI-Linked Proteins (biosynthesis)
• HeLa Cells
• Hep G2 Cells
• Histocompatibility Antigens Class I (biosynthesis, genetics)
• Humans
• Immunotherapy, Adoptive
• Interferon-gamma (biosynthesis, immunology)
• Interleukins (biosynthesis, genetics, pharmacology)
• Killer Cells, Natural (immunology)
• Lectins, C-Type (biosynthesis)
• Lymphocyte Activation (immunology)
• Membrane Glycoproteins (biosynthesis)
• NK Cell Lectin-Like Receptor Subfamily D (biosynthesis)
• NK Cell Lectin-Like Receptor Subfamily K (biosynthesis)
• Neoplasms (immunology, therapy)
• Receptors, IgG (biosynthesis)