title: 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.affiliation: Department of Biological Engineering, College of Pharmacy, Jilin University, 1266 Fu Jin Road, Changchun 130021, Jilin Province, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Jiang
authors.affiliation: First Hospital of Jilin University, Changchun 130021, Jilin Province, China.
authors.forename: Xuan
authors.initials: X
authors.lastname: Wu
authors.affiliation: Department of Endemic Diseases, Jilin University, Changchun 130021, Jilin Province, China.
authors.forename: Xi-Ning
authors.initials: XN
authors.lastname: Li
authors.affiliation: Department of Biological Engineering, College of Pharmacy, Jilin University, 1266 Fu Jin Road, Changchun 130021, Jilin Province, China.
authors.forename: Xi
authors.initials: X
authors.lastname: Yang
authors.affiliation: Department of Biological Engineering, College of Pharmacy, Jilin University, 1266 Fu Jin Road, Changchun 130021, Jilin Province, China.
authors.forename: Yulai
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Department of Biological Engineering, College of Pharmacy, Jilin University, 1266 Fu Jin Road, Changchun 130021, Jilin Province, China.
authors.forename: Haowei
authors.initials: H
authors.lastname: Yan
authors.affiliation: Department of Biological Engineering, College of Pharmacy, Jilin University, 1266 Fu Jin Road, Changchun 130021, Jilin Province, China.
authors.forename: An-Hui
authors.initials: AH
authors.lastname: Wei
authors.affiliation: Department of Biological Engineering, College of Pharmacy, Jilin University, 1266 Fu Jin Road, Changchun 130021, Jilin Province, China. Electronic address: WeiqunYan58@163.com.
authors.forename: Weiqun
authors.initials: W
authors.lastname: Yan
mesh_terms: D053330:4-1BB Ligand / Q000096:biosynthesis* / Q000235:genetics; D051997:ADP-ribosyl Cyclase 1 / Q000096:biosynthesis; D015703:Antigens, CD / Q000096:biosynthesis; D000945:Antigens, Differentiation, T-Lymphocyte / Q000096:biosynthesis; D019002:CD56 Antigen / Q000096:biosynthesis; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D058851:GPI-Linked Proteins / Q000096:biosynthesis; D006367:HeLa Cells; D056945:Hep G2 Cells; D015395:Histocompatibility Antigens Class I / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D016219:Immunotherapy, Adoptive; D007371:Interferon-gamma / Q000096:biosynthesis / Q000276:immunology; D007378:Interleukins / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D007694:Killer Cells, Natural / Q000276:immunology*; D037181:Lectins, C-Type / Q000096:biosynthesis; D008213:Lymphocyte Activation / Q000276:immunology; D008562:Membrane Glycoproteins / Q000096:biosynthesis; D051920:NK Cell Lectin-Like Receptor Subfamily D / Q000096:biosynthesis; D055655:NK Cell Lectin-Like Receptor Subfamily K / Q000096:biosynthesis; D009369:Neoplasms / Q000276:immunology* / Q000628:therapy; D017452:Receptors, IgG / Q000096:biosynthesis; D000098546:Interleukin-21; D000099192:CD69 Antigens
chemical_list: D053330:4-1BB Ligand; D051997:ADP-ribosyl Cyclase 1; D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D019002:CD56 Antigen; D058851:GPI-Linked Proteins; D015395:Histocompatibility Antigens Class I; D007371:Interferon-gamma; D007378:Interleukins; D037181:Lectins, C-Type; D008562:Membrane Glycoproteins; D051920:NK Cell Lectin-Like Receptor Subfamily D; D055655:NK Cell Lectin-Like Receptor Subfamily K; D017452:Receptors, IgG; D000098546:Interleukin-21; D000099192:CD69 Antigens; C428200:FCGR3B protein, human; C497869:KLRD1 protein, human; C527085:KLRK1 protein, human; C099187:MHC class I-related chain A; C522918:NCAM1 protein, human; C498154:CD38 protein, human
keywords: 4-1BBL; IL-21; K562 cells; MICA; NK cells