title: Real-time in vivo analysis of T cell activation in the central nervous system using a genetically encoded calcium indicator.
abstract: To study T cell activation in vivo in real time, we introduced a newly developed fluorescence resonance energy transfer-based, genetically encoded calcium indicator into autoantigen-specific and non-autoantigen-specific CD4(+) T cells. Using two-photon microscopy, we explored the responses of retrovirally transduced calcium indicator-expressing T cells to antigen in the lymph nodes and the central nervous system. In lymph nodes, the administration of exogenous antigen caused an almost immediate arrest of T cells around antigen-presenting cells and an instant rise of cytosolic calcium. In contrast, encephalitogenic T cells entering the leptomeningeal space, one main portal into the central nervous system parenchyma during experimental autoimmune encephalomyelitis, showed elevated intracellular calcium concentrations while still meandering through the space. This approach enabled us to follow the migration and activation patterns of T cells in vivo during the course of the disease.
authors.affiliation: Department of Neuroimmunology, Max Planck Institute of Neurobiology, Martinsried, Germany.
authors.forename: Marsilius
authors.initials: M
authors.lastname: Mues
authors.forename: Ingo
authors.initials: I
authors.lastname: Bartholomäus
authors.forename: Thomas
authors.initials: T
authors.lastname: Thestrup
authors.forename: Oliver
authors.initials: O
authors.lastname: Griesbeck
authors.forename: Hartmut
authors.initials: H
authors.lastname: Wekerle
authors.forename: Naoto
authors.initials: N
authors.lastname: Kawakami
authors.forename: Gurumoorthy
authors.initials: G
authors.lastname: Krishnamoorthy
mesh_terms: D000818:Animals; D017951:Antigen Presentation; D001921:Brain / Q000276:immunology* / Q000378:metabolism; D002118:Calcium / Q000032:analysis / Q000378:metabolism*; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D008198:Lymph Nodes / Q000378:metabolism; D008213:Lymphocyte Activation; D051379:Mice; D008810:Mice, Inbred C57BL; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism
chemical_list: D002118:Calcium