CXCL12+ fibroblastic reticular cells in lymph nodes facilitate immune tolerance

Lymph nodes (LNs) are the critical sites at which alloimmunity is activated. We have shown that the microenvironment within the LNs is a major determinant of the equilibrium between alloimmunity and suppression of the alloimmune response.
Fibroblastic reticular cells (FRCs) constitute a major subset of LN stromal cells (LNSCs). Recent data from our research indicate that T cells enter LNs in distinct ways depending on whether tolerance or immunity is being established, and they localize to specific domains within LNs.
CMFDA dye–stained CD3+ T cells were prepared to evaluate T cell trafficking. To visualize HEVs in LNs, 40 μg Alexa Fluor 647–conjugated anti–ER-TR7 antibody (Santa Cruz Biotechnology, sc-73355 AF647) was injected intravenously into WT C57BL/6J mice and Ccl19CreCxcl12fl/fl mice 2 hours before imaging. Mice were anesthetized […]. The popliteal LNs were exposed surgically and confocal intravital imaging was performed (IVM-CMS3, IVIM Technology). Twenty-five Z-stack images were obtained with a 3 μm axial interval. Time-lapse images were obtained at a 1-minute time interval for 20 minutes with 5 sequential Z-stack images of 3 μm axial interval.

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