De Brito T, Menezes L F, Lima Dirce M C, Lourenço Silvia, Silva Ana Maria G, Alves V A F
Institute of Tropical Medicine, University of Sao Paulo Medical School, Sao Paulo, SP, Brazil.
Virchows Arch. 2006 May;448(5):576-83. doi: 10.1007/s00428-006-0163-z. Epub 2006 Mar 16.
An in situ hybridization (ISH) assay for the detection of leptospiral DNA in tissues was described and its diagnostic and pathogenetic usefulness in combination with immunohistochemistry (IHC) was evaluated in formalin-fixed, paraffin-embedded liver and kidney samples from human fatal cases of leptospirosis. IHC assays with anti-E-cadherin antibodies assessed the liver-plate disarray frequently observed in leptospirosis. Immunohistochemistry detected leptospiral antigen (LAg) in macrophages, both in human liver and kidney. In guinea pigs, in addition to these findings, staining on cell membranes of hepatocytes and, occasionally, in apical membrane of kidney tubular cells was demonstrated. Positive ISH signal was observed chiefly in the nuclei of human hepatocytes and in the cytoplasm and nuclei of liver cells of experimentally infected guinea pigs. Loss of E-cadherin membrane expression is associated with liver-plate disarray. These findings were discussed in the contention that, in leptospirosis, cell membrane damage might be important for the pathogenesis of the disease. Finally, it was suggested that both IHC and/or ISH might be used for both diagnostic and research purposes.
描述了一种用于检测组织中钩端螺旋体DNA的原位杂交(ISH)检测方法,并在来自人类钩端螺旋体病致死病例的福尔马林固定、石蜡包埋的肝脏和肾脏样本中,评估了其与免疫组织化学(IHC)联合使用时的诊断和致病作用。用抗E-钙黏蛋白抗体进行的免疫组织化学检测评估了钩端螺旋体病中经常观察到的肝板紊乱情况。免疫组织化学在人类肝脏和肾脏的巨噬细胞中检测到了钩端螺旋体抗原(LAg)。在豚鼠中,除了这些发现外,还在肝细胞的细胞膜以及偶尔在肾小管细胞的顶膜上发现了染色。ISH阳性信号主要在人类肝细胞的细胞核以及实验感染豚鼠肝细胞的细胞质和细胞核中观察到。E-钙黏蛋白膜表达的缺失与肝板紊乱有关。在钩端螺旋体病中细胞膜损伤可能对疾病发病机制很重要这一论点下讨论了这些发现。最后,有人提出免疫组织化学和/或原位杂交可用于诊断和研究目的。