A new flow cytometric assay for the simultaneous analysis of antigen-specific elimination of T cells in heterogeneous T cell populations.
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.
Schütz
Christian
C
University of Regensburg, Department of Internal Medicine I, Franz-Josef-Strauss-Allee 11, 93042 Regensburg, Germany. christian.schuetz@klinik.uni-r.de
Fischer
Karin
K
Völkl
Simon
S
Hoves
Sabine
S
Halbritter
Dagmar
D
Mackensen
Andreas
A
Fleck
Martin
M