Heaney L G, Leggett P, Maxwell P, Bharucha H, Ennis M
Department of Respiratory Medicine, Royal Victoria Hospital, Belfast, Northern Ireland, UK.
Allergy. 1997 Aug;52(8):836-43. doi: 10.1111/j.1398-9995.1997.tb02155.x.
Reported mast-cell counts in endobronchial biopsies from asthmatic subjects are conflicting, with different methodologies often being used. This study compared three standard methods of counting mast cells in endobronchial biopsies from asthmatic and normal subjects. Endobronchial biopsies were obtained from atopic asthmatic subjects (n = 17), atopic nonasthmatic subjects (n = 6), and nonatopic nonasthmatic control subjects (n = 5). After overnight fixation in Carnoy's fixative, mast cells were stained by the short and long toluidine blue methods and antitryptase immunohistochemistry and were counted by light microscopy. Method comparison was made according to Bland & Altman. The limits of agreement were unacceptable for each of the comparisons, suggesting that the methods are not interchangeable. Coefficients of repeatability were excellent, and not different for the individual techniques. These results suggest that some of the reported differences in mast-cell numbers in endobronchial biopsies in asthma may be due to the staining method used, making direct comparisons between studies invalid. Agreement on a standard method is required for counting mast cells in bronchial biopsies, and we recommend the immunohistochemical method, since fixation is less critical and the resultant tissue sections facilitate clear, accurate, and rapid counts.
关于哮喘患者支气管活检中所报告的肥大细胞计数存在矛盾，这通常是由于使用了不同的方法。本研究比较了三种在哮喘患者和正常受试者支气管活检中计数肥大细胞的标准方法。支气管活检取自特应性哮喘患者（n = 17）、特应性非哮喘患者（n = 6）和非特应性非哮喘对照受试者（n = 5）。在卡诺伊固定液中过夜固定后，肥大细胞用短和长甲苯胺蓝法以及抗类胰蛋白酶免疫组织化学法染色，并通过光学显微镜计数。根据布兰德和奥特曼方法进行方法比较。每次比较的一致性界限都不可接受，这表明这些方法不可互换。重复性系数极佳，且各技术之间无差异。这些结果表明，哮喘患者支气管活检中所报告的肥大细胞数量差异部分可能归因于所使用的染色方法，这使得不同研究之间的直接比较无效。在支气管活检中计数肥大细胞需要就标准方法达成一致，我们推荐免疫组织化学方法，因为固定要求不那么严格，且所得组织切片便于清晰、准确和快速计数。
