Identification and Characterization of Tertiary Lymphoid Structures in Murine Melanoma.
Identification and Characterization of Tertiary Lymphoid Structures in Murine Melanoma.
复制标题
鼠黑色素瘤中三级淋巴结构的鉴定和表征。
DOI：
10.1007/978-1-4939-8709-2_14
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Engelhard VH
中科院分区：
文献类型：
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作者：
Rodriguez AB;Peske JD;Engelhard VH
三级淋巴样结构（TLS）是短暂的异位淋巴样聚集体，通常与传统的次级淋巴样器官具有结构相似性。在各种实体癌中，这些结构的存在通常与高密度的肿瘤浸润性T淋巴细胞和延长的患者生存期相关。这些观察结果表明，TLS作为有益的抗肿瘤免疫反应的发展的网站。然而，很少有小鼠肿瘤模型已经被描述，可以更全面地了解TLS在实体癌中的功能。我们以前报道过，小鼠B16-F1黑色素瘤或刘易斯肺癌细胞转染表达模型抗原卵清蛋白后形成肿瘤内TLS植入C57 BL/6小鼠腹腔。在这一章中，我们描述了免疫荧光显微镜和流式细胞术的方法来识别和表征肿瘤内TLS。此外，我们描述了一种过继转移方法，用于证明幼稚T细胞通过淋巴结样血管系统浸润到B16-OVA黑色素瘤肿瘤中，这是肿瘤相关TLS的基本功能特征。
Tertiary lymphoid structures (TLS) are transient ectopic lymphoid aggregates that often share structural similarities to conventional secondary lymphoid organs. In a variety of solid cancers, the presence of these structures commonly correlates with high densities of tumor-infiltrating T lymphocytes and prolonged patient survival. These observations suggest that TLS act as sites for the development of beneficial antitumor immune responses. However, few murine tumor models have been described that could enable a more comprehensive understanding of the functionality of TLS in solid cancers. We previously reported that murine B16-F1 melanoma or Lewis Lung carcinoma cells transfected to express the model antigen ovalbumin form intratumoral TLS after implantation into the peritoneal cavity of C57BL/6 mice. In this chapter, we describe immunofluorescent microscopy and flow cytometry approaches for identifying and characterizing intratumoral TLS. Additionally, we describe an adoptive transfer method for demonstrating the infiltration of naïve T cells into B16-OVA melanoma tumors via the lymph node-like vasculature that is an essential functional feature of tumor-associated TLS.