Snider Douglas B, Meyerholz David K, Dellon Evan S, Cortes Lizette M, Karri Akash, Blikslager Anthony T, Laster Scott, Käser Tobias, Cruse Glenn
Department of Molecular Biomedical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, United States.
Comparative Medicine Institute, North Carolina State University, Raleigh, NC, United States.
Front Vet Sci. 2025 Mar 19;12:1540995. doi: 10.3389/fvets.2025.1540995. eCollection 2025.
Accurate identification of eosinophils in tissue sections is required for diagnosis of eosinophilic esophagitis in humans and the assessment of severity of disease in allergy models. The pig may be a good model for sensitization and allergy models due to anatomical, physiological, and immunological similarities to humans. However, comparative studies on histochemical detection of eosinophils in fixed porcine tissue are lacking.
Qualitative and quantitative comparisons were performed for six histochemical methods previously reported for eosinophil and mast cell detection in other species. Astra Blue/Vital New Red, Congo Red, Luna, Sirius Red, Toluidine Blue, and modified regressive Hematoxylin & Eosin were applied to formalin-fixed paraffin embedded full-thickness sections of porcine esophagus. Specimens were collected from young, crossbred pigs sensitized to ovalbumin with or without subsequent oral exposure to ovalbumin to produce eosinophilic esophagitis lesions for comparison to non-allergic controls.
Ease of eosinophil quantitation was analyzed, and varied by histochemical stain, to determine whether stain selection increased accuracy and efficiency of evaluation. Noticeable differences in color contrast between intracytoplasmic granules, surrounding tissue, and cellular components aided detection and identification of eosinophils and mast cells with Astra Blue/New Vital Red and Toluidine Blue, respectively. For eosinophils, Congo Red and H&E were adequate, while Luna and Sirius Red presented challenges for quantitation.
In this case, rapid and reliable characterization of porcine esophageal allergy models was made possible by using Astra Blue/New Vital Red for eosinophils and Toluidine Blue for mast cells.
准确识别组织切片中的嗜酸性粒细胞对于人类嗜酸性粒细胞性食管炎的诊断以及过敏模型中疾病严重程度的评估至关重要。由于猪在解剖学、生理学和免疫学方面与人类相似，因此可能是致敏和过敏模型的良好选择。然而，目前缺乏关于固定猪组织中嗜酸性粒细胞组织化学检测的比较研究。
对先前报道的用于其他物种嗜酸性粒细胞和肥大细胞检测的六种组织化学方法进行了定性和定量比较。将阿斯特拉蓝/活性新红、刚果红、卢娜染色法、天狼星红、甲苯胺蓝和改良回归苏木精与伊红染色法应用于福尔马林固定石蜡包埋的猪食管全层切片。从对卵清蛋白致敏的年轻杂交猪中采集样本，无论是否随后口服卵清蛋白以产生嗜酸性粒细胞性食管炎病变，用于与非过敏对照进行比较。
分析了嗜酸性粒细胞定量的难易程度，其因组织化学染色而异，以确定染色选择是否提高了评估的准确性和效率。阿斯特拉蓝/新活性红和甲苯胺蓝分别使细胞质颗粒、周围组织和细胞成分之间的颜色对比度有明显差异，有助于嗜酸性粒细胞和肥大细胞的检测和识别。对于嗜酸性粒细胞，刚果红和苏木精与伊红染色法足够，但卢娜染色法和天狼星红染色法在定量方面存在挑战。
在这种情况下，通过使用阿斯特拉蓝/新活性红检测嗜酸性粒细胞和甲苯胺蓝检测肥大细胞，能够快速可靠地表征猪食管过敏模型。
