Monaco Sara E, Dacic Sanja, Seigh Lindsey, Hartman Douglas J, Xing Juan, Pantanowitz Liron
Department of Pathology, University of Pittsburgh Medical Center (UPMC), Pittsburgh, PA, USA.
Cytopathology. 2020 Sep;31(5):393-401. doi: 10.1111/cyt.12812. Epub 2020 Apr 6.
Immunotherapy has shown promising results in non-small cell lung cancer (NSCLC), for which tumour-infiltrating cytotoxic (CD8+) T cells play a critical role. We investigated the utility of image analysis (IA) to quantify CD8+ T cells in a series of matched small biopsies and resections of NSCLC.
CD8 immunohistochemistry was performed on cell-blocks (CB), core needle biopsies (CNB) and corresponding resections from primary NSCLCs. Slides were digitised using an Aperio AT2 scanner (Leica) and annotated by whole slide image (WSI) or fields of view occupied by tissue spots (TS). Quantitative IA was performed with a customised Aperio algorithm (Leica). CD8 scores (number of T cells with 1-3+ staining/total area) were then compared.
Forty-four cases with CB or CNB material and a corresponding resection were analysed. Average CD8 score was determined in CB (7.67 WSI, 77.67 TS) and/or CNB (47.35 WSI, 325.67 TS), and corresponding resections (190.35 WSI, 336.58 TS). CD8 score concordance was highest (78.6%) for CNBs using WSI annotation. Overall, small biopsies (CB or CNB) correlated with the resection in 71.4% cases using WSI and 63.3% cases using TS annotation. IA performed better for low CD8 scores.
These findings show that CD8 density in NSCLC can be quantified by IA in small biopsies and cell blocks, achieving the best concordance using WSI scores. Discrepancies were attributed to values near the cut-off and background detection of staining. These data warrant future studies with more cases and follow-up data to further investigate the clinical utility of IA for CD8 analysis in NSCLC.
免疫疗法在非小细胞肺癌（NSCLC）中已显示出有前景的结果，其中肿瘤浸润性细胞毒性（CD8 +）T细胞起着关键作用。我们研究了图像分析（IA）在一系列匹配的NSCLC小活检组织和手术切除组织中定量CD8 + T细胞的实用性。
对原发性NSCLC的细胞块（CB）、粗针活检（CNB）及相应手术切除组织进行CD8免疫组织化学检测。使用Aperio AT2扫描仪（徕卡）对玻片进行数字化处理，并通过全玻片图像（WSI）或组织斑点（TS）占据的视野进行标注。使用定制的Aperio算法（徕卡）进行定量IA。然后比较CD8评分（1 - 3 +染色的T细胞数量/总面积）。
分析了44例有CB或CNB材料及相应手术切除组织的病例。在CB（7.67 WSI，77.67 TS）和/或CNB（47.35 WSI，325.67 TS）以及相应手术切除组织（190.35 WSI，336.58 TS）中确定了平均CD8评分。使用WSI标注时，CNB的CD8评分一致性最高（78.6%）。总体而言，使用WSI标注时，71.4%的病例中小活检组织（CB或CNB）与手术切除组织相关，使用TS标注时为63.3%。IA对低CD8评分的表现更好。
这些发现表明，NSCLC中的CD8密度可通过IA在小活检组织和细胞块中进行定量，使用WSI评分时一致性最佳。差异归因于接近临界值的值和染色的背景检测。这些数据值得未来进行更多病例和随访数据的研究，以进一步探讨IA在NSCLC中进行CD8分析的临床实用性。
