Leclere Mathilde, Desnoyers Michel, Beauchamp Guy, Lavoie Jean-Pierre
Department of Clinical Sciences, Faculty of Veterinary Medicine, Université de Montréal, Saint-Hyacinthe, Québec, Canada.
J Vet Intern Med. 2006 Mar-Apr;20(2):377-81. doi: 10.1892/0891-6640(2006)20[377:cofsmf]2.0.co;2.
Mast cells normally are present in equine bronchoalveolar lavage fluid (BALF), but usually represent <2% of all cells in healthy horses. An increased percentage of mast cells has been associated with airway hyperactivity and inflammatory airway diseases, but marked differences are reported between studies in normal and diseased horses. Because an abnormal mast cell count may be of clinical relevance, we compared the ability of a fast Romanowsky method to stain mast cell granules with that of 3 metachromatic stains: automated Romanowsky, May-Grünwald Giemsa, and toluidine blue stains. The BALF cells from 24 horses were studied. A differential cell count was performed blindly on 400 cells. The percentages of mast cells obtained were analyzed by means of repeated-measures analysis of variance and Fischer's PLSD test. The Bland and Altman method was used to assess agreement among stains. The mean percentage of mast cells in BALF was significantly lower with the fast Romanowsky than with the automated Romanowsky, May-Grünwald Giemsa, and toluidine blue stains. With the fast Romanowsky stain, the metachromatic granules of mast cells were not stained, and their identification was based on morphologic criteria. Toluidine blue staining allowed detection of the highest mean percentage of mast cells, but was inadequate for performing a differential cell count on other cell types. In conclusion, fast Romanosky stain may be inadequate for detection of mast cells in equine BALF, whereas automated Romanowsky, May-Grünwald Giemsa, and toluidine blue stains provide metachromatic staining of mast cell granules.
肥大细胞通常存在于马的支气管肺泡灌洗液（BALF）中，但在健康马匹中通常占所有细胞的比例不到2%。肥大细胞百分比增加与气道高反应性和炎症性气道疾病有关，但正常和患病马匹的研究报告之间存在显著差异。由于异常的肥大细胞计数可能具有临床相关性，我们比较了快速罗曼诺夫斯基方法与三种异染性染色方法（自动罗曼诺夫斯基染色、美-姬氏染色和甲苯胺蓝染色）对肥大细胞颗粒染色的能力。对24匹马的BALF细胞进行了研究。对400个细胞进行了盲法细胞分类计数。通过重复测量方差分析和费舍尔最小显著差异检验分析获得的肥大细胞百分比。采用布兰德和奥特曼方法评估染色之间的一致性。快速罗曼诺夫斯基染色法检测到的BALF中肥大细胞的平均百分比显著低于自动罗曼诺夫斯基染色、美-姬氏染色和甲苯胺蓝染色法。使用快速罗曼诺夫斯基染色时，肥大细胞的异染性颗粒未被染色，其识别基于形态学标准。甲苯胺蓝染色能够检测到最高平均百分比的肥大细胞，但不足以对其他细胞类型进行细胞分类计数。总之，快速罗曼诺夫斯基染色可能不足以检测马BALF中的肥大细胞，而自动罗曼诺夫斯基染色、美-姬氏染色和甲苯胺蓝染色可对肥大细胞颗粒进行异染性染色。
