Menon Manisha, Benechet Alexandre P, Khanna Kamal M
Department of Immunology, University of Connecticut Health, 263 Farmington Ave, Farmington, CT, 06030, USA.
Division of Immunology, Transplantation and Infectious Diseases, IRCCS San Raffaele Scientific Institute, Via Olgettina 58, Milan, 20132, Italy.
Methods Mol Biol. 2017;1591:59-71. doi: 10.1007/978-1-4939-6931-9_5.
Local anatomy of lymphoid tissues during infection has emerged as a critical regulator of immunity; thus, studying the cellular choreography in the context of an intact tissue environment in situ is crucial. Following an infection, the local pathogen-specific T cell migration and the subsequent egress of effector T cells from the draining lymph nodes are important and complex biological processes. The mechanisms that regulate this complex process can now be investigated by directly visualizing T cell dynamics in vivo using intravital two-photon (2P) microscopy. In addition, static whole-mount imaging technique can provide us with a comprehensive assessment of global changes in the distribution of cellular populations within an intact tissue. Thus, in this chapter, we detail methods to visualize the migration and egress of endogenous antigen-specific CD8 T cells following viral infection using two methods-intravital 2P microscopy and multicolor whole-mount in situ tetramer staining.
感染期间淋巴组织的局部解剖结构已成为免疫的关键调节因子；因此，在完整组织环境中原位研究细胞编排至关重要。感染后，局部病原体特异性T细胞迁移以及随后效应T细胞从引流淋巴结的流出是重要且复杂的生物学过程。现在可以通过活体双光子（2P）显微镜在体内直接观察T细胞动态来研究调节这一复杂过程的机制。此外，静态整装成像技术可以为我们提供对完整组织内细胞群体分布的全局变化的全面评估。因此，在本章中，我们详细介绍了使用活体2P显微镜和多色整装原位四聚体染色这两种方法来观察病毒感染后内源性抗原特异性CD8 T细胞迁移和流出的方法。
