Siliņa Karīna, Burkhardt Chiara, Casanova Ruben, Solterman Alex, van den Broek Maries
Institute of Experimental Immunology, University of Zurich, Zurich, Switzerland.
Institute of Pathology and Molecular Pathology, University Hospital Zurich, Zurich, Switzerland.
Methods Mol Biol. 2018;1845:71-86. doi: 10.1007/978-1-4939-8709-2_5.
Tertiary lymphoid structures (TLS) develop in the human tumor microenvironment and correlate with prolonged survival in most cancer types. We recently demonstrated that TLS development follows sequential maturation stages and culminates in the generation of a germinal center (GC) reaction. This maturation process is crucial for the prognostic relevance of TLS in lung and colorectal cancer patients.The mechanisms underlying TLS development in various inflammatory conditions or their functional relevance in tumor immunity are not fully understood. Investigating which cell types and soluble mediators orchestrate lymphoid neogenesis in human tissues requires a method that allows simultaneous detection of multiple markers.Here, we describe a quantitative pathology approach to identify and quantify different TLS maturation stages in combination with other parameters. This approach consists of seven-color immunofluorescence protocol using tyramide signal amplification combined with multispectral microscopy and quantitative data acquisition from histological images.
三级淋巴结构（TLS）在人类肿瘤微环境中形成，并且在大多数癌症类型中与生存期延长相关。我们最近证明，TLS的形成遵循连续的成熟阶段，并最终产生生发中心（GC）反应。这一成熟过程对于TLS在肺癌和结直肠癌患者中的预后相关性至关重要。各种炎症条件下TLS形成的潜在机制或它们在肿瘤免疫中的功能相关性尚未完全了解。研究哪些细胞类型和可溶性介质协调人类组织中的淋巴新生需要一种能够同时检测多种标志物的方法。在此，我们描述了一种定量病理学方法，用于结合其他参数识别和量化不同的TLS成熟阶段。该方法包括使用酪胺信号放大的七色免疫荧光方案，结合多光谱显微镜和从组织学图像中获取定量数据。
