Department of Pathology, University of Oulu, FIN-90014 Oulu, Finland.
J Histochem Cytochem. 2010 Feb;58(2):131-40. doi: 10.1369/jhc.2009.954669.
We have optimized an immunohistochemical double-staining method combining immunohistochemical lymphocyte lineage marker detection and apoptosis detection with terminal deoxyribonucleotidyl transferase-mediated dUTP nick end labeling. The method was used to trace Fas-mediated apoptosis in human reactive lymph nodes according to cell lineage and anatomical location. In addition to Fas, we also studied the expression of Fas ligand (FasL), CD3, CD20, CD19, CD23, and CD68 of apoptotic cells. The presence of simultaneous Fas and FasL positivity indicated involvement of activation-induced death in the induction of paracortical apoptosis. FasL expression in the high endothelial venules might be an inductor of apoptosis of Fas-positive lymphoid cells. In addition to B-lymphocyte apoptosis in the germinal centers, there was often a high apoptosis rate of CD23-expressing follicular dendritic cells. In summary, our double-staining method provides valuable new information about the occurrence and mechanisms of apoptosis of different immune cell types in the lymph node compartments. Among other things, we present support for the importance of Fas/FasL-mediated apoptosis in lymph node homeostasis.
我们优化了一种免疫组织化学双重染色方法,该方法结合了免疫组织化学淋巴细胞谱系标志物检测和末端脱氧核糖核苷酸转移酶介导的 dUTP 缺口末端标记法检测凋亡。根据细胞谱系和解剖位置,该方法用于追踪 Fas 介导的人类反应性淋巴结中的细胞凋亡。除 Fas 外,我们还研究了凋亡细胞中 Fas 配体(FasL)、CD3、CD20、CD19、CD23 和 CD68 的表达。同时存在 Fas 和 FasL 阳性表明激活诱导的死亡参与了旁皮质凋亡的诱导。高内皮小静脉中 FasL 的表达可能是 Fas 阳性淋巴细胞凋亡的诱导剂。除生发中心的 B 淋巴细胞凋亡外,CD23 表达的滤泡树突状细胞的凋亡率通常也很高。总之,我们的双重染色方法为淋巴结隔室中不同免疫细胞类型的凋亡发生和机制提供了有价值的新信息。此外,我们还支持 Fas/FasL 介导的凋亡在淋巴结稳态中的重要性。