Punsoni Michael, Mangray Shamlal, Lombardo Kara A, Heath Nancy, Stopa Edward G, Yakirevich Evgeny
*Department of Pathology and Laboratory Medicine, Rhode Island Hospital †Warren Alpert Medical School of Brown University, Providence, RI.
Appl Immunohistochem Mol Morphol. 2017 Oct;25(9):645-650. doi: 10.1097/PAI.0000000000000432.
Succinate dehydrogenase (SDH) is a key mitochondrial enzyme complex composed of 4 subunits. SDH histochemistry is routinely utilized in the assessment of muscle biopsies to reveal underlying pathology such as subsarcolemmal mitochondrial aggregates. In this study, we evaluated the utility of succinate dehydrogenase B (SDHB) immunohistochemistry (IHC) in 27 muscle biopsies, including 13 mitochondrial myopathies (MMs), 9 inflammatory myopathies, and 5 controls. SDHB IHC was performed on formalin-fixed, paraffin-embedded tissue sections with a mouse monoclonal antibody (Abcam 21A11AE7) in parallel with histochemical SDH stains on a fresh-frozen tissue. In all muscle biopsies, SDHB IHC exhibited granular immunoreactivity and highlighted the dark type 1 and lighter type 2 staining pattern observed by histochemistry. In all cases of MM, SDHB IHC showed subsarcolemmal granular aggregates involving the entire periphery of the fibers that were more distinct than those seen by SDH histochemistry. In 3 extraocular muscle biopsies, SDHB immunoreactive speckles of various sizes were distributed throughout the entire sarcoplasm that were more prominent than those seen on SDH histochemistry. Subsarcolemmal and cytoplasmic granular aggregates seen on SDHB IHC correlated with mitochondrial pathology on electron microscopy. In cases of inflammatory myopathy, there was diffuse sarcoplasmic SDHB immunoreactivity in degenerating fibers, but no evidence of subsarcolemmal aggregates. This study demonstrates that SDHB IHC is highly sensitive and specific in the identification of MM. The automation, reproducibility, and cost efficiency of SDHB IHC offer advantages over the labor-intensive histochemical method requiring frozen sections. As this technique is performed on formalin-fixed, paraffin-embedded tissues, it can be easily applied for retrospective studies.
琥珀酸脱氢酶(SDH)是一种由4个亚基组成的关键线粒体酶复合物。SDH组织化学常用于评估肌肉活检,以揭示潜在的病理状况,如肌膜下线粒体聚集。在本研究中,我们评估了琥珀酸脱氢酶B(SDHB)免疫组织化学(IHC)在27例肌肉活检中的应用,其中包括13例线粒体肌病(MM)、9例炎性肌病和5例对照。使用小鼠单克隆抗体(Abcam 21A11AE7)对福尔马林固定、石蜡包埋的组织切片进行SDHB IHC检测,并与新鲜冷冻组织上的组织化学SDH染色同时进行。在所有肌肉活检中,SDHB IHC均表现出颗粒状免疫反应,并突出了组织化学观察到的深色1型和浅色2型染色模式。在所有MM病例中,SDHB IHC显示肌膜下颗粒聚集累及纤维的整个周边,比SDH组织化学所见更为明显。在3例眼外肌活检中,各种大小的SDHB免疫反应斑点分布于整个肌浆中,比SDH组织化学所见更为突出。SDHB IHC上观察到的肌膜下和细胞质颗粒聚集与电子显微镜下的线粒体病理相关。在炎性肌病病例中,变性纤维中有弥漫性肌浆SDHB免疫反应,但无肌膜下聚集的证据。本研究表明,SDHB IHC在MM的识别中具有高度敏感性和特异性。SDHB IHC的自动化、可重复性和成本效益优于需要冷冻切片的劳动密集型组织化学方法。由于该技术是在福尔马林固定、石蜡包埋的组织上进行的,因此可轻松应用于回顾性研究。