title: Identification and isolation of murine antigen-reactive T cells according to CD154 expression.
abstract: T helper (Th) cells are central regulators of adaptive immune responses. However, the detection of the small number of Th cells specific for a particular antigen or pathogen is still a major challenge. CD154 was recently introduced as a marker for antigen-specific Th cells. To date, this technology was not applicable for mice - arguably the most important immunological model system. CD154 is difficult to detect due to its rapid removal from the cell surface upon binding to CD40 during antigen-specific activation by APC. We present an efficient strategy to block the degradation of murine CD154 by combined use of antibodies against CD40 and CD154. This strategy makes CD154 easily accessible for surface staining, which allows isolation and expansion of rare antigen specific T cells. Importantly, CD154 identified all specific T cells in strongly Th1- or Th2-polarized immune responses against pathogens like Salmonella typhimurium and Heligmosomoides polygyrus, independent of their potential to produce cytokines. We demonstrate that CD154 can in fact be used as a reliable marker for antigen-specific CD4 T cells in mice, offering a unique option to analyze, isolate and rapidly expand the entire pool of Th-cells generated during a physiological T cell response in vivo.
authors: Immunomodulation Group, Deutsches Rheuma-Forschungszentrum Berlin, Berlin, Germany.
authors: Dennis
authors: D
authors: Kirchhoff
authors: Marco
authors: M
authors: Frentsch
authors: Patrick
authors: P
authors: Leclerk
authors: Dirk
authors: D
authors: Bumann
authors: Sebastian
authors: S
authors: Rausch
authors: Susanne
authors: S
authors: Hartmann
authors: Andreas
authors: A
authors: Thiel
authors: Alexander
authors: A
authors: Scheffold
mesh_terms: D000818:Animals; D019013:CD40 Antigens / Q000276:immunology / Q000378:metabolism; D023201:CD40 Ligand / Q000276:immunology* / Q000378:metabolism*; D002460:Cell Line; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D016207:Cytokines / Q000096:biosynthesis; D051379:Mice; D008807:Mice, Inbred BALB C; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology* / Q000378:metabolism*
chemical_list: D019013:CD40 Antigens; D016207:Cytokines; D023201:CD40 Ligand