Acute Lymph Node Slices Are a Functional Model System to Study Immunity Ex Vivo
Acute Lymph Node Slices Are a Functional Model System to Study Immunity Ex Vivo
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DOI：
10.1021/acsptsci.0c00143
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发表时间：
2021-01-08
影响因子：
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通讯作者：
Pompano，Rebecca R.
中科院分区：
文献类型：
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作者：
Belanger，Maura C.;Ball，Alexander G.;Pompano，Rebecca R.
淋巴结是一个高度组织化和动态的结构，对于促进构成适应性免疫的细胞间相互作用至关重要。淋巴结的大多数离体研究开始通过将其减少到细胞悬液，从而失去空间组织，或固定它，从而失去进行重复测量的能力。活鼠淋巴结组织切片提供了保留空间复杂性和动态可及性的潜力，但其活力、免疫激活水平和抗原特异性功能的保留尚未得到验证。在这里，我们系统地表征了活鼠淋巴结切片作为研究免疫的平台。活淋巴结切片保持了预期的空间组织和细胞群，同时反映了器官的3D空间复杂性。在优化条件下收集的切片在24小时活力和炎症方面与细胞悬液相当。切片对T细胞受体交联的反应是增加表面标志物表达和细胞因子分泌，在某些情况下比匹配的淋巴细胞培养物更强烈。此外，切片处理的蛋白抗原，和接种疫苗的动物的切片响应抗原特异性细胞因子分泌的离体挑战。总之，淋巴结切片提供了一个多功能的平台来研究空间组织中的免疫功能，从而实现明确的刺激、时程分析和并行读出。
The lymph node is a highly organized and dynamic structure that is critical for facilitating the intercellular interactions that constitute adaptive immunity. Most ex vivo studies of the lymph node begin by reducing it to a cell suspension, thus losing the spatial organization, or fixing it, thus losing the ability to make repeated measurements. Live murine lymph node tissue slices offer the potential to retain spatial complexity and dynamic accessibility, but their viability, level of immune activation, and retention of antigen-specific functions have not been validated. Here we systematically characterized live murine lymph node slices as a platform to study immunity. Live lymph node slices maintained the expected spatial organization and cell populations while reflecting the 3D spatial complexity of the organ. Slices collected under optimized conditions were comparable to cell suspensions in terms of both 24-h viability and inflammation. Slices responded to T cell receptor cross-linking with increased surface marker expression and cytokine secretion, in some cases more strongly than matched lymphocyte cultures. Furthermore, slices processed protein antigens, and slices from vaccinated animals responded to ex vivo challenge with antigen-specific cytokine secretion. In summary, lymph node slices provide a versatile platform to investigate immune functions in spatially organized tissue, enabling well-defined stimulation, time-course analysis, and parallel read-outs.