title: Mass Cytometry Analytical Approaches Reveal Cytokine-Induced Changes in Natural Killer Cells.
abstract: Natural killer (NK) cells have antiviral and antitumor activity that could be harnessed for the treatment of infections and malignancies. To maintain cell viability and enhance antiviral and antitumor effects, NK cells are frequently treated with cytokines. Here they performed an extensive assessment of the effects of cytokines on the phenotype and function of human NK cells.
  
  
  
  They used cytometry by time-of-flight (CyTOF) to evaluate NK cell repertoire changes after stimulation with interleukin (IL)-2, IL-15 or a combination of IL-12/IL-15/IL-18. To analyze the high dimensional CyTOF data, they used several statistical and visualization tools, including viSNE (Visualization of t-Distributed Stochastic Neighbor Embedding), Citrus (Cluster identification, characterization, and regression), correspondence analysis, and the Friedman-Rafsky test.
  
  
  
  All three treatments (IL-2, IL-15, and IL-12/IL-15/IL-18) increase expression of CD56 and CD69. The effects of treatment with IL-2 and IL-15 are nearly indistinguishable and characterized principally by increased expression of surface markers including CD56, NKp30, NKp44, and increased expression of functional markers, such as perforin, granzyme B, and MIP-1β. The combination of IL-12/IL-15/IL-18 induces a profound shift in the repertoire structure, decreasing expression of CD16, CD57, CD8, NKp30, NKp46, and NKG2D, and dramatically increasing expression of IFN-γ.
  
  
  
  CyTOF provides insights into the effects of cytokines on the phenotype and function of NK cells, which could inform future research efforts and approaches to NK cell immunotherapy. There are several analytical approaches to CyTOF data, and the appropriate method should be carefully selected based on which aspect of the dataset is being explored. © 2016 International Clinical Cytometry Society.
authors.affiliation: Department of Medicine, Stanford University, Stanford, California.
authors.forename: Elena
authors.initials: E
authors.lastname: Vendrame
authors.affiliation: Department of Statistics, Stanford University, Stanford, California.
authors.forename: Julia
authors.initials: J
authors.lastname: Fukuyama
authors.affiliation: Stanford Immunology, Stanford University, Stanford, California.
authors.forename: Dara M
authors.initials: DM
authors.lastname: Strauss-Albee
authors.affiliation: Department of Statistics, Stanford University, Stanford, California.
authors.forename: Susan
authors.initials: S
authors.lastname: Holmes
authors.affiliation: Department of Medicine, Stanford University, Stanford, California.|Stanford Immunology, Stanford University, Stanford, California.
authors.forename: Catherine A
authors.initials: CA
authors.lastname: Blish
mesh_terms: D016207:Cytokines / Q000378:metabolism*; D005434:Flow Cytometry / Q000379:methods; D053804:Granzymes / Q000378:metabolism; D006801:Humans; D016130:Immunophenotyping; D007167:Immunotherapy / Q000379:methods; D007694:Killer Cells, Natural / Q000166:cytology* / Q000276:immunology; D055629:Natural Cytotoxicity Triggering Receptor 2 / Q000276:immunology; D054353:Perforin / Q000378:metabolism
chemical_list: D016207:Cytokines; C112837:NCR2 protein, human; D055629:Natural Cytotoxicity Triggering Receptor 2; D054353:Perforin; D053804:Granzymes
keywords: IL-15; IL-2; cytokines; mass cytometry; memory-like natural killer cells; natural killer cells