Pulmonary Research Institute at Hospital Grosshansdorf, Center for Pneumology and Thoracic Surgery, Grosshansdorf, Germany.
Respir Med. 2013 Apr;107(4):587-95. doi: 10.1016/j.rmed.2012.12.006. Epub 2013 Jan 10.
Induced sputum is a non-invasive sampling technique for the analysis of airway inflammation in various lung diseases and comprises valuable potential for the identification of biomarkers and therapeutic targets by molecular methods. In the context of biobanking with preservation of induced sputum samples for subsequent analyses we applied the HEPES-glutamic acid buffer-mediated organic solvent protection effect (HOPE)-technique for preparation of induced sputum samples. Induced sputum samples of 20 patients with moderate to severe chronic obstructive pulmonary disease (COPD) and 12 healthy controls were collected. Cell pellets of induced sputum samples were preserved with HOPE and subsequently embedded in paraffin. Immunostaining of paraffin-block sections for interleukin-8, interleukin-17, myeloperoxidase, matrixmetalloproteinase-9, CD68, and CD8 revealed distinct signals without antigen retrieval. Moreover, RNA was extracted and successfully used for transcription microarray analysis. Sputum samples preserved by the HOPE-technique display a tool to address scientific approaches in pulmonary research, which can enable the identification of new biomarkers and therapeutic targets in respiratory diseases.
诱导痰是一种非侵入性采样技术，可用于分析各种肺部疾病中的气道炎症，通过分子方法具有鉴定生物标志物和治疗靶点的巨大潜力。在为后续分析保存诱导痰样本的生物库背景下，我们应用 HEPES-谷氨酸缓冲介导的有机溶剂保护效应 (HOPE) 技术来制备诱导痰样本。收集了 20 名中重度慢性阻塞性肺疾病 (COPD) 患者和 12 名健康对照者的诱导痰样本细胞沉淀，并用 HOPE 保存，随后嵌入石蜡。对石蜡块切片进行白细胞介素-8、白细胞介素-17、髓过氧化物酶、基质金属蛋白酶-9、CD68 和 CD8 的免疫染色显示出无需抗原修复的明显信号。此外，还提取了 RNA，并成功用于转录微阵列分析。通过 HOPE 技术保存的痰液样本提供了一种工具，可以解决肺部研究中的科学方法，从而能够确定呼吸疾病中的新生物标志物和治疗靶点。
