人类黑色素瘤中三级淋巴结构的标准化分析:与生发中心改变相关的疾病进展和肿瘤部位相关的变化。

A Standardized Analysis of Tertiary Lymphoid Structures in Human Melanoma: Disease Progression- and Tumor Site-Associated Changes With Germinal Center Alteration.

机构信息

Laboratory of Molecular Dermato-Oncology and Tumor Immunology, Department of Dermatology, Medical University of Vienna, Vienna, Austria.

Institute of Medical Genetics and Pathology, University Hospital Basel, University Basel, Basel, Switzerland.

出版信息

Front Immunol. 2021 Jun 24;12:675146. doi: 10.3389/fimmu.2021.675146. eCollection 2021.

Abstract

There is increasing evidence that tertiary lymphoid structures (TLS) control not only local adaptive B cell responses at melanoma tumor sites but also the cellular composition and function of other immune cells. In human melanoma, however, a comprehensive analysis of TLS phenotypes, density and spatial distribution at different disease stages is lacking. Here we used 7-color multiplex immunostaining of whole tissue sections from 103 human melanoma samples to characterize TLS phenotypes along the expression of established TLS-defining molecular and cellular components. TLS density and spatial distribution were determined by referring TLS counts to the tissue area within defined intra- and extratumoral perimeters around the invasive tumor front. We show that only a subgroup of primary human melanomas contains TLS. These TLS rarely formed germinal centers and mostly located intratumorally within 1 mm distance to the invasive tumor front. In contrast, melanoma metastases had a significantly increased density of secondary follicular TLS. They appeared preferentially in stromal areas within an extratumoral 1 mm distance to the invasive tumor front and their density varied over time and site of metastasis. Interestingly, secondary follicular TLS in melanoma often lacked BCL6 lymphatic cells and canonical germinal center polarity with the formation of dark and light zone areas. Our work provides an integrated qualitative, quantitative and spatial analysis of TLS in human melanoma and shows disease progression- and site-associated changes in TLS phenotypes, density and spatial distribution. The frequent absence of canonical germinal center polarity in melanoma TLS highlights the induction of TLS maturation as a potential additive to future immunotherapy studies. Given the variable evaluation strategies used in previous TLS studies of human tumors, an important asset of this study is the standardized quantitative evaluation approach that provides a high degree of reproducibility.

摘要

越来越多的证据表明,三级淋巴结构(TLS)不仅控制着黑色素瘤肿瘤部位局部适应性 B 细胞的反应,还控制着其他免疫细胞的细胞组成和功能。然而,在人类黑色素瘤中,缺乏对 TLS 表型、不同疾病阶段的密度和空间分布的全面分析。在这里,我们使用 7 色多重免疫组织化学染色法对 103 个人类黑色素瘤样本的全组织切片进行分析,以描述 TLS 表型,同时参考已建立的 TLS 定义分子和细胞成分的表达情况。TLS 密度和空间分布通过将 TLS 计数与浸润性肿瘤前缘周围定义的肿瘤内和肿瘤外边界内的组织面积进行比较来确定。我们发现只有一小部分原发性人类黑色素瘤含有 TLS。这些 TLS 很少形成生发中心,大多数位于肿瘤内,距离浸润性肿瘤前缘 1mm 以内。相比之下,黑色素瘤转移瘤的次级滤泡性 TLS 密度显著增加。它们优先出现在浸润性肿瘤前缘 1mm 以外的基质区域,其密度随时间和转移部位而变化。有趣的是,黑色素瘤中的次级滤泡性 TLS 常缺乏 BCL6 淋巴样细胞和经典的生发中心极性,形成暗区和亮区。我们的工作对人类黑色素瘤中的 TLS 进行了综合的定性、定量和空间分析,并显示了疾病进展和转移部位相关的 TLS 表型、密度和空间分布的变化。黑色素瘤 TLS 中经典生发中心极性的频繁缺失突出了 TLS 成熟的诱导作为未来免疫治疗研究的潜在附加因素。鉴于以前人类肿瘤 TLS 研究中使用的评估策略各不相同,本研究的一个重要优势是采用了标准化的定量评估方法,具有高度的可重复性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6006/8264652/98cd93495016/fimmu-12-675146-g001.jpg

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