Department of Biopathology, Institut Bergonié, Bordeaux, France; University of Bordeaux, Bordeaux, France.
Centre de Recherche des Cordeliers, Institut national de la santé et de la recherche médicale (INSERM), Université Sorbonne-Paris-Cité, Paris, France.
Lab Invest. 2023 May;103(5):100063. doi: 10.1016/j.labinv.2023.100063. Epub 2023 Jan 18.
Mature tertiary lymphoid structures (mTLSs) are organized lymphoid structures containing B lymphocytes admixed to CD23+ follicular dendritic cells. Their presence has been linked to improved survival and sensitivity to immune checkpoint inhibitors in several cancers, emerging as a promising pancancer biomarker. However, the requirements for any biomarker are clear methodology, proven feasibility, and reliability. In 357 patients' samples, we studied tertiary lymphoid structures (TLSs) parameters using multiplex immunofluorescence (mIF), hematoxylin-eosin-saffron (HES) staining, double CD20/CD23 staining, and single CD23 immunohistochemistry. The cohort included carcinomas (n = 211) and sarcomas (n = 146), gathering biopsies (n = 170), and surgical specimens (n = 187). mTLSs were defined as TLSs containing either a visible germinal center on HES staining or CD23+ follicular dendritic cells. Focusing on 40 TLSs assessed using mIF, double CD20/CD23 staining was less sensitive than mIF to assess maturity in 27.5% (n = 11/40) but was rescued by single CD23 staining in 90.9% (n = 10/11). In 97 patients, several samples (n = 240) were reviewed to characterize TLS distribution. The likelihood of finding TLSs in surgical material was 6.1 higher than in biopsy and 2.0 higher in primary samples than in metastasis after adjustment with a type of sample. Interrater agreement rates over 4 examiners were 0.65 (Fleiss kappa, 95% CI [0.46, 0.90]) for the presence of TLS and 0.90 for maturity (95% CI [0.83, 0.99]). In this study, we propose a standardized method to screen mTLSs in cancer samples using HES staining and immunohistochemistry that can be applied to all specimens.
成熟的三级淋巴样结构（mTLS）是一种含有 B 淋巴细胞的淋巴样组织，与 CD23+滤泡树突状细胞混合。在几种癌症中，其存在与生存率提高和对免疫检查点抑制剂的敏感性提高有关，成为一种有前途的泛癌生物标志物。然而，任何生物标志物的要求都很明确，需要有明确的方法、已被证实的可行性和可靠性。在 357 名患者的样本中，我们使用多重免疫荧光（mIF）、苏木精-伊红-藏红花（HES）染色、双 CD20/CD23 染色和单 CD23 免疫组化研究了三级淋巴样结构（TLS）的参数。该队列包括癌（n=211）和肉瘤（n=146），包括活检（n=170）和手术标本（n=187）。mTLS 被定义为含有 HES 染色可见生发中心或 CD23+滤泡树突状细胞的 TLS。在使用 mIF 评估的 40 个 TLS 中，双 CD20/CD23 染色的敏感性比 mIF 低 27.5%（n=11/40），但通过单 CD23 染色可挽救 90.9%（n=10/11）。在 97 名患者中，对多个样本（n=240）进行了回顾性分析，以描述 TLS 的分布。在调整了样本类型后，与活检相比，手术标本中发现 TLS 的可能性高 6.1 倍，与转移相比，原发标本中发现 TLS 的可能性高 2.0 倍。在 4 名检查者中，TLS 存在的组内一致性率为 0.65（Fleiss kappa，95%CI[0.46，0.90]），TLS 成熟度的组内一致性率为 0.90（95%CI[0.83，0.99]）。在这项研究中，我们提出了一种使用 HES 染色和免疫组化筛选癌症样本中 mTLS 的标准化方法，该方法可应用于所有标本。
