BACKGROUND New and fully validated tests need to be brought into clinical practice to improve the estimation of recurrence risk in patients with colon cancer. The aim of this study was to assess the analytical performances of the Immunoscore (IS) and show its contribution to prognosis prediction. METHODS Immunohistochemical staining of CD3+ and CD8+ T cells on adjacent sections of colon cancer tissues were quantified in the core of the tumor and its invasive margin with dedicated IS modules integrated into digital pathology software. Staining intensity across samples collected between 1989 and 2016 (n=595) was measured. The accuracy of the IS workflow was established by comparing optical and automatic counts. Analytical precision of the IS was evaluated within individual tumor block on distant sections and between eligible blocks. The IS interlaboratory reproducibility (n=100) and overall assay precision were assessed (n=3). Contribution of the IS to prediction of recurrence based on clinical and molecular parameters was determined (n=538). RESULTS Optical and automatic counts for CD3+ or CD8+ were strongly correlated (r=0.94, p<0.001 and r=0.92, p<0.001, respectively). CD3 and CD8 staining intensities were not altered by the age of the tumor block over a period of 30 years. Neither the position of tested tissue sections within a tumor block nor the selection of the tissue blocks affected the IS. Reproducibility of the IS was not affected by multiple variables (eg, antibody lots, DAB revelation kits, immunohistochemistry automates and operators). Interassay repeatability of the IS was 100% and interlaboratory reproducibility between two testing centers was 93%. Finally, in a case series of patients with stage II-III colon cancer, the relative proportion of variance for time to recurrence was greatest for the IS (53% of prognostic variability) in a model that included IS, T-stage, microsatellite instability status and total number of lymph nodes. CONCLUSION IS is a robust and validated clinical assay leveraging immune scoring to predict recurrence risk of patient with localized colon cancer. The strong and independent prognostic value of IS should pave the way for it use in clinical practice.

中文翻译：

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结肠癌患者免疫评分及其相关预后价值的分析验证。

背景 需要将新的和经过充分验证的测试引入临床实践，以改进对结肠癌患者复发风险的估计。本研究的目的是评估免疫评分 (IS) 的分析性能并展示其对预后预测的贡献。方法使用集成到数字病理学软件中的专用 IS 模块，在肿瘤核心及其浸润边缘量化结肠癌组织相邻切片上 CD3+ 和 CD8+ T 细胞的免疫组织化学染色。测量了 1989 年至 2016 年（n = 595）之间收集的样本的染色强度。IS 工作流程的准确性是通过比较光学计数和自动计数来确定的。在远处切片上的单个肿瘤块内和合格块之间评估 IS 的分析精度。评估了 IS 实验室间重现性 (n=100) 和整体检测精度 (n=3)。确定了基于临床和分子参数的 IS 对复发预测的贡献（n = 538）。结果 CD3+ 或 CD8+ 的光学计数和自动计数具有很强的相关性（分别为 r=0.94，p<0.001 和 r=0.92，p<0.001）。在 30 年的时间里，CD3 和 CD8 染色强度不会因肿瘤块的年龄而改变。肿瘤块内测试组织切片的位置和组织块的选择都不影响 IS。IS 的重现性不受多个变量（例如，抗体批次、DAB 揭示试剂盒、免疫组织化学自动化和操作员）的影响。IS 的批间重复性为 100%，两个测试中心之间的实验室间重复性为 93%。最后，在 II-III 期结肠癌患者的病例系列中，在包括 IS、T 分期、微卫星不稳定性的模型中，IS（预后变异性的 53%）的复发时间相对方差比例最大淋巴结的状态和总数。结论 IS 是一种强大且经过验证的临床测定，利用免疫评分来预测局部结肠癌患者的复发风险。IS 强大且独立的预后价值应为其在临床实践中的应用铺平道路。结论 IS 是一种强大且经过验证的临床测定，利用免疫评分来预测局部结肠癌患者的复发风险。IS 强大且独立的预后价值应为其在临床实践中的应用铺平道路。结论 IS 是一种强大且经过验证的临床测定，利用免疫评分来预测局部结肠癌患者的复发风险。IS 强大且独立的预后价值应为其在临床实践中的应用铺平道路。