Department of Anatomical Pathology, Faculty of Medicine, Vajira Hospital, Navamindradhiraj University, 681 Samsen Road, Dusit, Bangkok, 10300, Thailand.
Virchows Arch. 2019 Jan;474(1):87-96. doi: 10.1007/s00428-018-2458-2. Epub 2018 Sep 29.
Mitotic figure (MF) counting is important in the evaluation of meningioma grading. Nevertheless, mitosis assessment on hematoxylin and eosin (H&E)-stained slides may be problematic because of technical factors and pathologist's experience. Phosphohistone H3 (PHH3) is a mitosis-specific antibody that has proven to facilitate mitotic count in various tumors. However, the antibody performance between PHH3 serine10 (S10) and serine28 (S28) has never been compared in these tumors before. In this study, 48 cases of meningioma (28 grade I, 14 grade II, 6 grade III) were evaluated using immunohistochemical stains for four commercially available PHH3 (S10) and S28 antibodies to identify MFs and validate PHH3 intra- and interobserver reproducibility and agreement. Two pathologists counted MFs on both H&E- and PHH3-stained slides. H&E and PHH3 MFs were highly correlated (Spearman's rho = 0.96 for PHH3 (S10)-Biocare, 0.96 for PHH3 (S10)-CST, 0.91 for PHH3 (S28)-Abcam, and 0.89 for PHH3 (S28)-Santa Cruz. The mean difference between an H&E and PHH3 mitotic count is 0.81 for PHH3 (S10)-Biocare, 0.95 for PHH3 (S10)-CST, - 0.97 for PHH3 (S28)-Abcam, and - 0.97 for PHH3 (S28)-Santa Cruz. For comparison among four PHH3 antibodies, PHH3 mitotic counts had both a good intra- and interobserver reproducibility (p > 0.05). Regarding to World Health Organization (WHO) grade, there was not a significant discrepancy in the stratification of tumor grades for all four PHH3 antibodies in terms of interobserver agreement. The Cohen's kappa coefficient (K) was 0.93 for PHH3 (S10)-Biocare, 0.82 for PHH3 (S10)-CST, 0.76 for PHH3 (S28)-Abcam, and 0.80 for PHH3 (S28)-Santa Cruz. Considering survival analyses, all five proliferation indices were univariately associated with recurrences. Increased PHH3 mitotic indices (MIs) were significantly associated with recurrence-free survival in univariate Cox proportional hazards regression analysis (p < 0.001) and remained an independent predictor in multivariate analysis (p < 0.05). The appropriate prognostic cutoff values for recurrence prediction were 5 or more per 10 high-power fields (HPFs) for PHH3 (S10) and 3 or more per 10 HPFs for PHH3 (S28).
有丝分裂计数在脑膜瘤分级评估中很重要。然而，由于技术因素和病理学家的经验，在苏木精和伊红（H&E）染色切片上评估有丝分裂可能存在问题。磷酸组蛋白 H3（PHH3）是一种有丝分裂特异性抗体，已被证明可促进各种肿瘤中的有丝分裂计数。然而，在这些肿瘤中，PHH3 丝氨酸 10（S10）和丝氨酸 28（S28）之间的抗体性能从未进行过比较。在这项研究中，使用免疫组织化学染色评估了 48 例脑膜瘤（28 级 I、14 级 II、6 级 III），使用四种市售的 PHH3（S10）和 S28 抗体来鉴定有丝分裂，并验证 PHH3 内和观察者之间的可重复性和一致性。两位病理学家在 H&E 和 PHH3 染色切片上计数有丝分裂。H&E 和 PHH3 有丝分裂高度相关（PHH3（S10）-Biocare 的 Spearman rho = 0.96，PHH3（S10）-CST 的 Spearman rho = 0.96，PHH3（S28）-Abcam 的 Spearman rho = 0.91，PHH3（S28）-Santa Cruz 的 Spearman rho = 0.89）。H&E 和 PHH3 有丝分裂计数之间的平均差异为 PHH3（S10）-Biocare 为 0.81，PHH3（S10）-CST 为 0.95，PHH3（S28）-Abcam 为-0.97，PHH3（S28）-Santa Cruz 为-0.97。为了比较四种 PHH3 抗体，PHH3 有丝分裂计数具有良好的观察者内和观察者间可重复性（p>0.05）。关于世界卫生组织（WHO）分级，在四种 PHH3 抗体的观察者间一致性方面，肿瘤分级的分层没有显著差异。PHH3（S10）-Biocare 的 Cohen's kappa 系数（K）为 0.93，PHH3（S10）-CST 的 K 为 0.82，PHH3（S28）-Abcam 的 K 为 0.76，PHH3（S28）-Santa Cruz 的 K 为 0.80。考虑到生存分析，所有五个增殖指数均与复发相关。在单变量 Cox 比例风险回归分析中，PHH3 有丝分裂指数（MI）增加与无复发生存显著相关（p<0.001），并且在多变量分析中仍然是独立的预测因子（p<0.05）。用于预测复发的适当预后截断值为 PHH3（S10）每 10 个高倍视野（HPF）超过 5 个，PHH3（S28）每 10 个 HPF 超过 3 个。
