Humphries Matthew P, Bingham Victoria, Abdullahi Sidi Fatima, Craig Stephanie G, McQuaid Stephen, James Jacqueline, Salto-Tellez Manuel
Precision Medicine Centre of Excellence, The Patrick G Johnston Centre for Cancer Research, Queen's University, Belfast BT9 7BL, UK.
Cellular Pathology, Belfast Health and Social Care Trust, Belfast City Hospital, Lisburn Road, Belfast BT9 7BL, UK.
Cancers (Basel). 2020 Apr 29;12(5):1114. doi: 10.3390/cancers12051114.
Targeting of the programmed cell death protein (PD-1)/programmed death-ligand 1 (PD-L1) axis with checkpoint inhibitors has changed clinical practice in non-small cell lung cancer (NSCLC). However, clinical assessment remains complex and ambiguous. We aim to assess whether digital image analysis (DIA) and multiplex immunofluorescence can improve the accuracy of PD-L1 diagnostic testing. A clinical cohort of routine NSCLC patients reflex tested for PD-L1 (SP263) immunohistochemistry (IHC), was assessed using DIA. Samples of varying assessment difficulty were assessed by multiplex immunofluorescence. Sensitivity, specificity, and concordance was evaluated between manual diagnostic evaluation and DIA for chromogenic and multiplex IHC. PD-L1 expression by DIA showed significant concordance (R² = 0.8248) to manual assessment. Sensitivity and specificity was 86.8% and 91.4%, respectively. Evaluation of DIA scores revealed 96.8% concordance to manual assessment. Multiplexing enabled PD-L1+/CD68+ macrophages to be readily identified within PD-L1+/cytokeratin+ or PD-L1-/cytokeratin+ tumor nests. Assessment of multiplex vs. chromogenic IHC had a sensitivity and specificity of 97.8% and 91.8%, respectively. Deployment of DIA for PD-L1 diagnostic assessment is an accurate process of case triage. Multiplex immunofluorescence provided higher confidence in PD-L1 assessment and could be offered for challenging cases by centers with appropriate expertise and specialist equipment.
使用检查点抑制剂靶向程序性细胞死亡蛋白（PD-1）/程序性死亡配体1（PD-L1）轴已改变了非小细胞肺癌（NSCLC）的临床实践。然而，临床评估仍然复杂且不明确。我们旨在评估数字图像分析（DIA）和多重免疫荧光是否能提高PD-L1诊断检测的准确性。对接受PD-L1（SP263）免疫组织化学（IHC）检测的常规NSCLC患者的临床队列进行DIA评估。通过多重免疫荧光评估不同评估难度的样本。评估了手动诊断评估与DIA在显色和多重IHC方面的敏感性、特异性和一致性。DIA检测的PD-L1表达与手动评估显示出显著一致性（R² = 0.8248）。敏感性和特异性分别为86.8%和91.4%。对DIA评分的评估显示与手动评估的一致性为96.8%。多重检测能够在PD-L1+/细胞角蛋白+或PD-L1-/细胞角蛋白+肿瘤巢内轻松识别PD-L1+/CD68+巨噬细胞。多重IHC与显色IHC的评估敏感性和特异性分别为97.8%和91.8%。采用DIA进行PD-L1诊断评估是一种准确的病例分类方法。多重免疫荧光为PD-L1评估提供了更高的可信度，具备适当专业知识和专业设备的中心可将其用于具有挑战性的病例。
