Bastan Idil, Rendahl Aaron K, Seelig Davis, Day Michael J, Hall Edward J, Rao Savita P, Washabau Robert J, Sriramarao P
Department of Veterinary Clinical Sciences, University of Minnesota, St. Paul, Minnesota.
Department of Veterinary and Biomedical Sciences, University of Minnesota, St. Paul, Minnesota.
J Vet Intern Med. 2018 Nov;32(6):1911-1917. doi: 10.1111/jvim.15310. Epub 2018 Oct 7.
Accurate identification of eosinophils in the gastrointestinal (GI) tract of dogs with eosinophilic GI disease (EGID) by histological evaluation is challenging. The currently used hematoxylin and eosin (H&E) staining method detects intact eosinophils but does not detect degranulated eosinophils, thus potentially underrepresenting the number of infiltrating eosinophils.
To develop a more sensitive method for identifying and quantifying both intact and degranulated eosinophils to diagnose EGID more accurately.
Endoscopically obtained paraffin-embedded intestinal biopsy specimens from dogs with GI signs were examined. The study groups were dogs with eosinophilic enteritis (EE), lymphoplasmacytic and mixed enteritis, and control dogs with GI signs but no histologic changes on tissue sections. Consecutive sections were immunolabeled with monoclonal antibodies (mAbs) against the eosinophil granule protein eosinophil peroxidase (Epx) and stained by H&E, respectively. The number of eosinophils was manually quantified and classified as intact or degranulated.
The number of intact eosinophils detected in Epx mAb-labeled duodenal sections was significantly higher compared with that in H&E-stained sections, with a similar relationship noted in the colon and stomach. The Epx mAb allowed the unique assessment of eosinophil degranulation. The number of intact and degranulated eosinophils was significantly higher in duodenal lamina propria of the EE and mixed group compared to the control group.
Immunohistochemical detection of Epx provides a more precise method to detect GI tract eosinophils compared to H&E staining and could be used as an alternative and reliable diagnostic tool for assessment of biopsy tissues from dogs with EGID.
通过组织学评估准确识别嗜酸性胃肠道疾病（EGID）犬胃肠道（GI）中的嗜酸性粒细胞具有挑战性。目前使用的苏木精和伊红（H&E）染色方法可检测完整的嗜酸性粒细胞，但无法检测脱颗粒的嗜酸性粒细胞，因此可能低估了浸润嗜酸性粒细胞的数量。
开发一种更敏感的方法来识别和定量完整及脱颗粒的嗜酸性粒细胞，以更准确地诊断EGID。
检查了经内镜获取的有胃肠道症状犬的石蜡包埋肠活检标本。研究组包括嗜酸性肠炎（EE）、淋巴浆细胞性和混合性肠炎的犬，以及有胃肠道症状但组织切片无组织学变化的对照犬。连续切片分别用抗嗜酸性粒细胞颗粒蛋白嗜酸性粒细胞过氧化物酶（Epx）的单克隆抗体（mAb）进行免疫标记，并用H&E染色。手动定量嗜酸性粒细胞的数量，并将其分类为完整或脱颗粒。
与H&E染色切片相比，Epx mAb标记的十二指肠切片中检测到的完整嗜酸性粒细胞数量显著更高，在结肠和胃中也观察到类似的关系。Epx mAb允许对嗜酸性粒细胞脱颗粒进行独特评估。与对照组相比，EE组和混合组十二指肠固有层中完整和脱颗粒嗜酸性粒细胞的数量显著更高。
与H&E染色相比，Epx的免疫组织化学检测提供了一种更精确的方法来检测胃肠道嗜酸性粒细胞，可作为评估EGID犬活检组织的一种替代且可靠的诊断工具。
