Abstract
Upon antigen recognition, Tcells form either static (synapses) or migratory (kinapses) contacts with antigen-presenting cells. Addressing whether synapses and kinapses result in distinct Tcell receptor (TCR) signals has been hampered by the inability to simultaneously assess Tcell phenotype and behavior. Here, we introduced dynamic insitu cytometry (DISC), a combination of intravital multiphoton imaging and flow cytometry-like phenotypic analysis. Taking advantage of CD62L shedding as a marker of early TCR signaling, we examined how Tcells sense TCR ligands of varying affinities invivo. We uncovered three modes of antigen recognition: synapses with the strongest TCR signals, kinapses with robust signaling, and kinapses with weak signaling. As illustrated here, the DISC approach should provide unique opportunities to link immune cell behavior to phenotype and function invivo.
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