title: A novel, dual cytokine-secretion assay for the purification of human Th22 cells that do not co-produce IL-17A.
abstract: BACKGROUND
  Interleukin-22 is produced by certain T helper cells subsets (Th17, Th22) and at lower levels by γ-δ T cells, NKT and innate lymphoid cells. Th22 cells are unique immune cells that regulate tissue responses by IL-22 production. The exact discrimination between Th17 cells that co-produce IL-22 and single IL-22-producing Th22 cells has not been possible until the present study. Isolation of pure Th22 cells without co-expression of cytokines of other T-cell subsets is essential to better understand their function in humans. The aim of this study is the isolation and characterization of viable, human IL-22-producing CD4+ T cells that do not produce IL-17A.
  
  
  METHODS
  Isolation of viable Th22 cells was performed with the combination of two cytokine secretion assays detecting IL-17A- and IL-22-producing cells in a single purification step.
  
  
  RESULTS
  The newly developed cytokine secretion assay consists of anti-IL-22 and anti-IL-17A catch antibodies, which via biotin-streptavidin interaction are bound to the biotinylated surface of the target cell, and anti-IL-22 and IL-17A detection antibody labelled with a fluorescent dye, which detects cytokines bound to these catch antibodies. A unique population of human Th22 cells, which do not produce IL-17A, was sorted, and cytokine expression pattern was confirmed by quantitative PCR analysis and ELISA. The presented technique allows the detection and isolation of pure human Th22 cells.
  
  
  CONCLUSIONS
  This technique may allow the purification of any single cytokine-producing cell subset, and the combination of several different cytokine secretion assays can be used to purify and characterize novel and unique cell subsets.
authors.affiliation: Swiss Institute of Allergy and Asthma Research (SIAF), University of Zurich, Davos, Switzerland.
authors.forename: M
authors.initials: M
authors.lastname: Wawrzyniak
authors.affiliation: Swiss Institute of Allergy and Asthma Research (SIAF), University of Zurich, Davos, Switzerland.
authors.forename: U
authors.initials: U
authors.lastname: Ochsner
authors.affiliation: Swiss Institute of Allergy and Asthma Research (SIAF), University of Zurich, Davos, Switzerland.
authors.forename: O
authors.initials: O
authors.lastname: Wirz
authors.affiliation: Swiss Institute of Allergy and Asthma Research (SIAF), University of Zurich, Davos, Switzerland.|CK-CARE AG - Christine Kühne - Center for Allergy Research and Education, Davos, Switzerland.
authors.forename: P
authors.initials: P
authors.lastname: Wawrzyniak
authors.affiliation: Swiss Institute of Allergy and Asthma Research (SIAF), University of Zurich, Davos, Switzerland.
authors.forename: W
authors.identifier: http://orcid.org/0000-0001-9951-6688
authors.initials: W
authors.lastname: van de Veen
authors.affiliation: Swiss Institute of Allergy and Asthma Research (SIAF), University of Zurich, Davos, Switzerland.|CK-CARE AG - Christine Kühne - Center for Allergy Research and Education, Davos, Switzerland.
authors.forename: C A
authors.initials: CA
authors.lastname: Akdis
authors.affiliation: Swiss Institute of Allergy and Asthma Research (SIAF), University of Zurich, Davos, Switzerland.
authors.forename: M
authors.initials: M
authors.lastname: Akdis
mesh_terms: D000954:Antigens, Surface / Q000378:metabolism; D002469:Cell Separation / Q000379:methods; D016207:Cytokines / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D020381:Interleukin-17 / Q000096:biosynthesis; D007378:Interleukins / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016176:T-Lymphocyte Subsets / Q000378:metabolism*; D000097043:Interleukin-22
chemical_list: D000954:Antigens, Surface; D016207:Cytokines; D020381:Interleukin-17; D007378:Interleukins; D012333:RNA, Messenger; D000097043:Interleukin-22
keywords: Interleukin-17A; Interleukin-22; cytokine-secretion assay; human Th17 cells; human Th22 cells