Alimentiv Inc.
STTARR Innovation Core Facility, Princess Margaret Cancer Centre, University Health Network.
Appl Immunohistochem Mol Morphol. 2022 Aug 1;30(7):486-492. doi: 10.1097/PAI.0000000000001035. Epub 2022 May 20.
The T-lymphocyte-mediated inflammation in Crohn's disease can be assessed by quantifying CD3-positive T-lymphocyte counts in colonic sections. We developed and validated a process to reliably quantify immunohistochemical marker-positive cells in a high-throughput setting using whole slide images (WSIs) of CD3-immunostained colonic and ileal tissue sections. In regions of interest (ROIs) and/or whole tissue sections of 40 WSIs from 36 patients with Crohn's disease, CD3-positive cells were quantified by an expert gastrointestinal pathologist (gold standard) and by image analysis algorithms developed with software from 3 independent vendors. Semiautomated quantification of CD3-positive cell counts estimated in 1 ROI per section were accurate when compared with manual analysis (Pearson correlation coefficient, 0.877 to 0.925). Biological variability was acceptable in digitally determined CD3-positive cell measures between 2 to 5 ROIs annotated on the same tissue section (coefficient of variation <25%). Results from computer-aided analysis of CD3-positive T lymphocytes in a whole tissue section and the average of results from 2 to 5 ROIs per tissue section lacked reliability (overestimation or underestimation and systematic bias), suggesting that absolute quantification of CD3-positive T lymphocytes in a whole tissue section may be more accurate. Semiautomated image analysis in WSIs demonstrated reproducible CD3-positive cell measures across 3 independent algorithms. A computer-aided digital image analysis method was developed and validated to quantify CD3-positive T lymphocytes in colonic and ileal biopsy sections from patients with Crohn's disease. Results support consideration of this digital analysis method for use in future Crohn's disease clinical studies.
T 淋巴细胞介导的克罗恩病炎症可以通过定量结肠切片中 CD3 阳性 T 淋巴细胞计数来评估。我们开发并验证了一种在高通量环境中使用 CD3 免疫染色的结肠和回肠组织切片的全幻灯片图像（WSI）可靠地定量免疫组织化学标志物阳性细胞的方法。在 36 名克罗恩病患者的 40 张 WSI 的感兴趣区域（ROI）和/或整个组织切片中，由一位专家胃肠病理学家（金标准）和 3 个独立供应商的软件开发的图像分析算法对 CD3 阳性细胞进行定量。与手动分析相比，对每个切片 1 个 ROI 进行的 CD3 阳性细胞计数的半自动定量是准确的（Pearson 相关系数为 0.877 至 0.925）。在同一组织切片上注释的 2 到 5 个 ROI 之间，数字化确定的 CD3 阳性细胞测量值的生物学变异性可接受（变异系数 <25%）。在整个组织切片中对 CD3 阳性 T 淋巴细胞进行计算机辅助分析的结果以及每个组织切片 2 到 5 个 ROI 的结果平均值缺乏可靠性（高估或低估和系统偏差），这表明整个组织切片中 CD3 阳性 T 淋巴细胞的绝对定量可能更准确。WSI 中的半自动图像分析显示了 3 种独立算法之间具有可重复性的 CD3 阳性细胞测量值。已经开发并验证了一种计算机辅助数字图像分析方法，用于定量克罗恩病患者的结肠和回肠活检切片中的 CD3 阳性 T 淋巴细胞。结果支持在未来的克罗恩病临床研究中考虑使用这种数字分析方法。
