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TitlePhotoconversion of Alloreactive T Cells in Murine Peyer’s Patches During Acute Graft-Versus-Host Disease: Tracking the Homing Route of Highly Proliferative Cells In Vivo
Author Jarick, Mokhtari, Scheller, Hartweg, Thusek, Le, Ranecky, Shaikh, Qureischi, Heinze and Beilhack
Affiliation(s)University of Würzburg, Würzburg, Germany
PublishedFront Immunol. 2018; 9: 1468. doi: [10.3389/fimmu.2018.01468]
KeywordSilver LED-405 nm T cell migration, acute graft-versus-host disease, mouse models, photoconversion, Dendra2, Peyer’s patch, in vivo cell tracking, lymphocyte homing
Snippet High-power UV LED lamp, fiber-coupled, Silver LED-405 nm, Prizmatix.
AbstractThe regulation of immune cell migration throughout the body is essential to warrant immunosurveillance and to maintain immune homeostasis. Marking and tracking of these cells has proven important to study mechanisms of immune cell trafficking and cell interaction in vivo. Photoconversion is a well-suited technique for intravital application because it enables contactless time- and location-specific marking of cells in the tissue without surgically manipulating the microenvironment of the cells in question. However, in dividing cells the converted fluorescent protein may decline quickly. Here, we provide a detailed description of the photoconversion technique and its applicability to tracking highly proliferating T cells from the priming site of T cell activation to peripheral target organs of effector function in a preclinical model. Dendra2+ T cells were photoconverted in the Peyer’s patches during the initiation phase of acute graft-versus-host disease (GvHD) and tracked through the mesenteric lymph nodes and the peripheral blood to the small intestine with flow cytometry and intravital two-photon microscopy. Photoconverted alloreactive T cells preserved the full proliferative capacity, homing, and migration of alloreactive T cells in the intestinal lamina propria......

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