title: A new flow cytometric assay for the simultaneous analysis of antigen-specific elimination of T cells in heterogeneous T cell populations.
abstract: Novel immunosuppressive strategies are targeting for an antigen-specific deletion of T cells responsible for organ damage in autoimmunity and allograft rejection. Here, we present a new flow cytometry-based assay that allows the reliable and efficient detection of T cells that were eliminated in an antigen-specific fashion. A stable cell-labelling technique utilizing the two membrane dyes PKH26 and PKH67 has been combined with annexin V and 7-aminoactinomycin (7-AAD) staining to detect apoptotic cells. A differential gating strategy enabled us to determine the viability/apoptosis for each PKH-stained T cell subpopulation independently. The capability to simultaneously analyze apoptosis within T cell mixtures of different antigen specificities establishes this assay as a superior tool for the further development of novel antigen-specific immunosuppressive approaches.
authors.affiliation: University of Regensburg, Department of Internal Medicine I, Franz-Josef-Strauss-Allee 11, 93042 Regensburg, Germany. christian.schuetz@klinik.uni-r.de
authors.forename: Christian
authors.initials: C
authors.lastname: Schütz
authors.forename: Karin
authors.initials: K
authors.lastname: Fischer
authors.forename: Simon
authors.initials: S
authors.lastname: Völkl
authors.forename: Sabine
authors.initials: S
authors.lastname: Hoves
authors.forename: Dagmar
authors.initials: D
authors.lastname: Halbritter
authors.forename: Andreas
authors.initials: A
authors.lastname: Mackensen
authors.forename: Martin
authors.initials: M
authors.lastname: Fleck