Department of Pathology, Clínica Universidad de Navarra, Pamplona, Spain.
Centro de Investigación Biomédica en Red de Cáncer (CIBERONC), Madrid, Spain.
Cytopathology. 2021 Sep;32(5):611-616. doi: 10.1111/cyt.12979. Epub 2021 Apr 18.
Understanding the immune environment of non-small cell lung cancer (NSCLC) is important for designing effective anticancer immunotherapies. We describe the use of multiplex immunofluorescence (mIF) assays to enable characterisation of the tumour-infiltrating immune cells and their interactions, both across and within immune subtypes.
Six cytological samples of NSCLC taken by transoesophageal ultrasound-guided fine needle aspiration were tested with an mIF assay designed to detect the expression of key immune cell markers such as CD3, CD8, CD20, CD11b, and CD68. Pan-cytokeratin was used to detect the NSCLC cells. Fluorescence images were acquired on a Vectra-Polaris Automated Quantitative Pathology Imaging System (Akoya Biosciences).
MIF assay was able to reliably detect and quantify the myeloid cell markers CD11b, CD68, CD3+ and CD8+ T cells, and CD20+ B lymphocytes on cytological samples of NSCLC. Whole-tissue analysis and its correlation with the corresponding H&E stains allowed a better understanding of the tissue morphology and the relationship between tumour and stroma compartments. Additionally, a uniform, specific, and correct staining pattern was seen for every immune marker.
The implementation of mIF assay on cytological samples taken with minimally invasive methods seems feasible and can be used to explore the immune environment of NSCLC.
了解非小细胞肺癌（NSCLC）的免疫环境对于设计有效的抗癌免疫疗法至关重要。我们描述了使用多重免疫荧光（mIF）检测来描述肿瘤浸润免疫细胞及其相互作用，包括在免疫亚型内和跨免疫亚型的特征。
对经经食管超声引导下细针抽吸术获取的 6 例 NSCLC 细胞学样本进行 mIF 检测，该检测旨在检测关键免疫细胞标志物的表达，如 CD3、CD8、CD20、CD11b 和 CD68。细胞角蛋白 pan 用于检测 NSCLC 细胞。荧光图像在 Vectra-Polaris 自动化定量病理学成像系统（Akoya Biosciences）上获取。
mIF 检测能够可靠地检测和定量 NSCLC 细胞学样本中的髓样细胞标志物 CD11b、CD68、CD3+和 CD8+T 细胞以及 CD20+B 淋巴细胞。对整个组织进行分析，并与相应的 H&E 染色进行相关性分析，有助于更好地了解组织形态和肿瘤与基质隔室之间的关系。此外，每种免疫标志物的染色模式均具有一致性、特异性和正确性。
采用微创方法获取的细胞学样本上实施 mIF 检测似乎是可行的，并可用于探索 NSCLC 的免疫环境。
