Bastan Idil, Robinson Nicholas A, Ge Xiao Na, Rendahl Aaron K, Rao Savita P, Washabau Robert J, Sriramarao P
Am J Vet Res. 2017 Jan;78(1):36-41. doi: 10.2460/ajvr.78.1.36.
OBJECTIVE To evaluate a method for identifying intact and degranulated eosinophils in the small intestine of dogs with inflammatory bowel disease (IBD) by use of a monoclonal antibody (mAb) against eosinophil peroxidase (EPX). ANIMALS 11 untreated dogs with IBD, 5 dogs with IBD treated with prednisolone, and 8 control dogs with no clinical evidence of gastrointestinal tract disease and no immunosuppressive treatment. PROCEDURES 4-μm-thick sections of paraffin-embedded tissues from necropsy specimens were immunostained with EPX mAb. Stained intact and degranulated eosinophils in consecutive microscopic fields (400X magnification) of the upper (villus tips) and lower (between the muscularis mucosae and crypts) regions of the lamina propria of the jejunum were manually counted. RESULTS Compared with control and treated IBD dogs, untreated IBD dogs had a significantly higher number of degranulated eosinophils in the lower region of the lamina propria. However, no significant differences were detected in the number of intact eosinophils in this region among groups. In the upper region of the lamina propria, untreated IBD dogs had a significantly higher number of degranulated and intact eosinophils, compared with control and treated IBD dogs. Number of degranulated and intact eosinophils did not differ significantly between control and treated IBD dogs. CONCLUSIONS AND CLINICAL RELEVANCE Immunohistologic analysis with EPX mAb yielded prominent granule staining that allowed reliable morphological identification of degranulated and intact eosinophils, which may provide a strategy for quantitative and selective evaluation of eosinophils in gastrointestinal biopsy specimens and a potential method to diagnose IBD and evaluate treatment outcome.
目的 评估一种通过使用抗嗜酸性粒细胞过氧化物酶(EPX)的单克隆抗体(mAb)来识别患有炎症性肠病(IBD)的犬小肠中完整和脱颗粒嗜酸性粒细胞的方法。动物 11 只未经治疗的患有 IBD 的犬、5 只接受泼尼松龙治疗的患有 IBD 的犬以及 8 只无胃肠道疾病临床证据且未接受免疫抑制治疗的对照犬。方法 对尸检标本石蜡包埋组织的 4μm 厚切片用 EPX mAb 进行免疫染色。在空肠固有层上部(绒毛尖端)和下部(黏膜肌层与隐窝之间)区域的连续显微镜视野(400 倍放大)中手动计数染色的完整和脱颗粒嗜酸性粒细胞。结果 与对照犬和接受治疗的 IBD 犬相比,未经治疗的 IBD 犬固有层下部区域的脱颗粒嗜酸性粒细胞数量显著更高。然而,各组在该区域完整嗜酸性粒细胞数量上未检测到显著差异。在固有层上部区域,与对照犬和接受治疗的 IBD 犬相比,未经治疗的 IBD 犬脱颗粒和完整嗜酸性粒细胞数量显著更高。对照犬和接受治疗的 IBD 犬之间脱颗粒和完整嗜酸性粒细胞数量无显著差异。结论及临床意义 用 EPX mAb 进行免疫组织学分析产生了显著的颗粒染色,可对脱颗粒和完整嗜酸性粒细胞进行可靠的形态学识别,这可能为胃肠道活检标本中嗜酸性粒细胞的定量和选择性评估提供一种策略,以及一种诊断 IBD 和评估治疗结果的潜在方法。