Sammicheli Stefano, Kuka Mirela, Iannacone Matteo
Division of Immunology, Transplantation and Infectious Diseases, IRCCS San Raffaele Scientific Institute, Milan, Italy.
Vita-Salute San Raffaele University, Milan, Italy.
Methods Mol Biol. 2018;1763:63-74. doi: 10.1007/978-1-4939-7762-8_7.
Humoral immune responses depend on B cells encountering antigen (Ag) in lymph nodes (LNs) draining infection sites, getting activated, interacting with different cells, proliferating and differentiating into antibody (Ab)-secreting cells. Each of these events occurs in distinct LN sub-compartments, requiring the migration of B cells from niche to niche in a fast and tightly coordinated fashion. While some of the rules that characterize B cell behavior in secondary lymphoid organs have been elucidated at the population level, we have only limited knowledge of the precise dynamics of B cell interactions with different kinds of LN cells at the single-cell level. Here, we describe in detail an intravital microscopy technique that allows the analysis of B cell dynamic behavior in the popliteal lymph node of anesthetized mice at high spatial and temporal resolution. A detailed understanding of the spatiotemporal dynamics of B cells within secondary lymphoid organs may lead to novel, rational vaccine strategies aimed at inducing rapid and long-lived humoral immune responses.
体液免疫反应依赖于B细胞在引流感染部位的淋巴结（LN）中遇到抗原（Ag），被激活，与不同细胞相互作用，增殖并分化为分泌抗体（Ab）的细胞。这些事件中的每一个都发生在不同的LN亚区，需要B细胞以快速且紧密协调的方式从一个生态位迁移到另一个生态位。虽然在群体水平上已经阐明了一些表征B细胞在次级淋巴器官中行为的规则，但我们对B细胞在单细胞水平上与不同类型LN细胞相互作用的精确动态了解有限。在这里，我们详细描述了一种活体显微镜技术，该技术能够以高空间和时间分辨率分析麻醉小鼠腘淋巴结中B细胞的动态行为。对次级淋巴器官内B细胞时空动态的详细了解可能会带来旨在诱导快速和持久体液免疫反应的新型合理疫苗策略。
