Bollampalli Vishnu Priya, Nylén Susanne, Rothfuchs Antonio Gigliotti
Department of Microbiology, Tumor and Cell Biology (MTC), Karolinska Institutet.
Department of Microbiology, Tumor and Cell Biology (MTC), Karolinska Institutet;
J Vis Exp. 2016 Oct 9(116):54620. doi: 10.3791/54620.
Dendritic cells (DCs) are important for initiating immune responses, in part through their ability to acquire and shuttle antigen to the draining lymph node (DLN). The mobilization of DCs to the DLN is complex and remains to be fully elucidated during infection. Herein described is the use of an innovative, simple assay that relies on the fluorochrome 5- and 6-carboxyfluorescein diacetate succinimidyl ester (CFSE) to track the migration of DCs during footpad infection with Mycobacterium bovis Bacille Calmette-Guérin (BCG) in C57BL/6 mice. This assay enables the characterization of skin DC sub-populations that actively relocate to the draining, popliteal LN in response to BCG. This protocol originates from a BCG model where migratory skin DCs were identified by flow cytometry. The assay is amiable to the study and identification of DCs or other cells that home to the popliteal LN after inoculation of microbes, their metabolites or other inflammatory stimuli in the footpad, and consequently to study factors that regulate the migration of these cells.
树突状细胞（DCs）对于启动免疫反应很重要，部分原因在于它们获取抗原并将其转运至引流淋巴结（DLN）的能力。DCs向DLN的动员过程很复杂，在感染期间仍有待充分阐明。本文描述了一种创新的简单检测方法，该方法依赖于荧光染料5-和6-羧基荧光素二乙酸琥珀酰亚胺酯（CFSE）来追踪C57BL/6小鼠在足垫感染卡介苗（BCG）期间DCs的迁移情况。该检测方法能够对响应BCG而主动迁移至引流的腘窝淋巴结的皮肤DC亚群进行表征。该方案源自一个BCG模型，在该模型中通过流式细胞术鉴定了迁移性皮肤DCs。该检测方法适用于研究和鉴定在足垫接种微生物、其代谢产物或其他炎症刺激后归巢至腘窝淋巴结的DCs或其他细胞，从而研究调节这些细胞迁移的因素。
