Redefining Histological Cell Counts Using a Standardized Method: The Leuven Intestinal Counting Protocol.

IF 3 3区医学 Q2 GASTROENTEROLOGY & HEPATOLOGY Clinical and Translational Gastroenterology Pub Date : 2024-07-01 DOI:10.14309/ctg.0000000000000725

Matthias Ceulemans, Pauline Huyghe, Gert De Hertogh, Raquel Cameron, Jolien Schol, Grace L Burns, Simon Keely, Lucas Wauters, Jan Tack, Nicholas J Talley, Tim Vanuytsel

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Abstract

Introduction: The diagnosis of eosinophilic gastrointestinal diseases is largely based on mucosal eosinophil counts, but thresholds and normal ranges beyond the esophagus are debated, calling for much-needed methodological standardization. We aimed to develop a standardized workflow for duodenal cell quantification and estimate duodenal eosinophil and mast cell numbers in healthy controls.

Methods: Software-based histological cell quantification using free-sized or fixed-sized regions was developed and applied to digitized hematoxylin and eosin (H&E)-stained slides from 58 individuals (healthy controls [HCs] and patients with functional dyspepsia). Intraclass correlation coefficients (ICCs) compared inter-rater reliability between software-based and microscopic quantification. Reproducibility of the software-based method was validated in an independent cohort of 37 control and functional dyspepsia subjects. Eosinophil identification on H&E staining was compared to immunohistochemistry (IHC). Normal eosinophil (H&E) and mast cell (cKit) ranges were determined in 70 adult HCs.

Results: Eosinophil quantification on digitized slides demonstrated excellent (ICC = 0.909) and significantly improved reproducibility over microscopic evaluation (ICC = 0.796, P = 0.0014), validated in an independent cohort (ICC = 0.910). Duodenal eosinophils were more abundant around crypts than in villi ( P < 0.0001), while counts were similar on matched H&E- and IHC-stained slides ( P = 0.55). Mean ± SD (95th percentile) duodenal eosinophils and mast cells in HC were 228.8/mm 2 ± 94.7 (402.8/mm 2 ) and 419.5/mm 2 ± 132.2 (707.6/mm 2 ), respectively.

Discussion: We developed and validated a standardized approach to duodenal histological cell quantification, generalizable to various mucosal cell types. Implementation of software-based quantification identified 400 eosinophils/mm 2 and 700 mast cells/mm 2 as thresholds for abnormal duodenal infiltration.

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使用标准化方法重新定义组织细胞计数：鲁汶肠道计数方案。

目的：嗜酸性粒细胞胃肠道疾病的诊断主要基于粘膜嗜酸性粒细胞计数，但对食管以外的阈值和正常范围存在争议，因此亟需方法标准化。我们旨在开发十二指肠细胞定量的标准化工作流程，并估算健康对照组的十二指肠嗜酸性粒细胞和肥大细胞数量：方法：我们开发了基于软件的组织学细胞定量方法，使用自由大小或固定大小的区域，并将其应用于来自 58 人（健康对照组（HC）和功能性消化不良（FD）患者）的数字化苏木精和伊红（H&E）染色玻片。类内相关系数（ICC）比较了基于软件的量化和显微镜量化之间的相互可靠性。基于软件的方法的再现性在 37 名对照组和 FD 受试者的独立队列中得到了验证。嗜酸性粒细胞的H&E染色鉴定与免疫组化（IHC）进行了比较。在 70 名成年 HC 中确定了嗜酸性粒细胞（H&E）和肥大细胞（cKit）的正常范围：结果：数字化切片上的嗜酸性粒细胞定量结果显示极佳（ICC：0.909），与显微镜评估（ICC：0.796，P = .0014）相比，可重复性显著提高，这在一个独立的队列中得到验证（ICC：0.910）。十二指肠嗜酸性粒细胞在隐窝周围比在绒毛中更丰富（P < .0001），而在匹配的 H&E 和 IHC 染色切片上计数相似（P = .55）。HC中十二指肠嗜酸性粒细胞和肥大细胞的平均值±标准偏差（第95百分位数）分别为228.8/mm2±94.7（402.8/mm2）和419.5/mm2±132.2（707.6/mm2）：我们开发并验证了十二指肠组织学细胞定量的标准化方法，该方法适用于各种粘膜细胞类型。基于软件的定量方法确定了 400 个嗜酸性粒细胞/mm2 和 700 个肥大细胞/mm2 作为十二指肠异常浸润的阈值。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Clinical and Translational Gastroenterology GASTROENTEROLOGY & HEPATOLOGY-

CiteScore

7.00

自引率

0.00%

发文量

114

审稿时长

16 weeks

期刊介绍： Clinical and Translational Gastroenterology (CTG), published on behalf of the American College of Gastroenterology (ACG), is a peer-reviewed open access online journal dedicated to innovative clinical work in the field of gastroenterology and hepatology. CTG hopes to fulfill an unmet need for clinicians and scientists by welcoming novel cohort studies, early-phase clinical trials, qualitative and quantitative epidemiologic research, hypothesis-generating research, studies of novel mechanisms and methodologies including public health interventions, and integration of approaches across organs and disciplines. CTG also welcomes hypothesis-generating small studies, methods papers, and translational research with clear applications to human physiology or disease. Colon and small bowel Endoscopy and novel diagnostics Esophagus Functional GI disorders Immunology of the GI tract Microbiology of the GI tract Inflammatory bowel disease Pancreas and biliary tract Liver Pathology Pediatrics Preventative medicine Nutrition/obesity Stomach.