Biomarkers may hold the key towards development and improvement of personalized cancer treatment. For instance, tumour expression of immune system-related proteins may reveal the tumour immune status and, accordingly, determine choice for type of immunotherapy. Therefore, objective evaluation of tumour biomarker expression is needed but often challenging. For instance, human leukocyte antigen (HLA) class I tumour epithelium expression is cumbersome to quantify by eye due to its presence on both tumour epithelial cells and tumour stromal cells, as well as tumour-infiltrating immune cells. In this study, we solved this problem by setting up an immunohistochemical (IHC) double staining using a tissue microarray (TMA) of rectal tumours wherein HLA class I expression was coloured with a blue chromogen, whereas non-epithelial tissue was visualized with a brown chromogen. We subsequently developed a semi-automated image analysis method that identified tumour epithelium as well as the percentage of HLA class I-positive tumour epithelium. Using this technique, we compared HCA2/HC10 and EMR8-5 antibodies for the assessment of HLA class I tumour expression and concluded that EMR8-5 is the superior antibody for this purpose. This IHC double staining can in principle be used for scoring of any biomarker expressed by tumour epithelium.

中文翻译：

---

一种直肠癌中HLA I类肿瘤上皮表达的半自动图像分析方法。

生物标志物可能是发展和改善个性化癌症治疗的关键。例如，免疫系统相关蛋白的肿瘤表达可揭示肿瘤的免疫状态，并因此确定免疫疗法类型的选择。因此，需要对肿瘤生物标志物表达进行客观评估，但通常具有挑战性。例如，人白细胞抗原（HLA）I类肿瘤上皮表达由于在肿瘤上皮细胞和肿瘤基质细胞以及浸润肿瘤的免疫细胞中均存在而难以通过眼睛进行定量。在这项研究中，我们通过使用直肠肿瘤的组织微阵列（TMA）进行免疫组织化学（IHC）双重染色来解决此问题，其中HLA I类表达被蓝色色原染色，非上皮组织则显示为棕色色原。随后，我们开发了一种半自动图像分析方法，该方法可识别肿瘤上皮细胞以及HLA I类阳性肿瘤上皮细胞的百分比。使用该技术，我们比较了HCA2 / HC10和EMR8-5抗体对HLA I类肿瘤表达的评估，并得出结论，EMR8-5是用于此目的的优越抗体。这种IHC双重染色原则上可用于对肿瘤上皮表达的任何生物标志物进行评分。我们比较了HCA2 / HC10和EMR8-5抗体对HLA I类肿瘤表达的评估，并得出结论，EMR8-5是用于此目的的优越抗体。这种IHC双重染色原则上可用于对肿瘤上皮表达的任何生物标志物进行评分。我们比较了HCA2 / HC10和EMR8-5抗体对HLA I类肿瘤表达的评估，并得出结论，EMR8-5是用于此目的的优越抗体。这种IHC双重染色原则上可用于对肿瘤上皮表达的任何生物标志物进行评分。