Wiwien H W, Hiyama K, Maeda H, Ishioka S, Yamakido M
Second Department of Internal Medicine, Hiroshima University School of Medicine, Japan.
Hiroshima J Med Sci. 1996 Mar;45(1):1-10.
Sarcoidosis is a systemic granulomatous disease of unknown origin. To clarify its pathogenesis, we searched for known or unknown genes which are specifically expressed in sarcoidosis. Bronchoalveolar lavage (BAL) cells from 18 patients with sarcoidosis and 8 patients with various lung diseases were analyzed by differential display method. mRNA was extracted from BAL cells and reverse transcribed with 12 kinds of anchored primer, which theoretically cover all mRNAs, followed by polymerase chain reaction (PCR) with the anchored primer and a 10-mer arbitrary primer. PCR products were displayed on a polyacrylamide gel and fragments showing characteristic alterations in intensity between sarcoidosis and other patients were extracted, sequenced, and compared against Genbank and EMBL DNA data bases. One fragment was detected with specifically increased intensity and another disappeared in patients with sarcoidosis. These fragments were likely derived from unknown genes. CD44 and tumor necrosis factor (TNF-alpha) cDNA sequences were also detected as fragments commonly expressed in sarcoidosis. The cloned fragments with specifically increased or decreased intensity in sarcoidosis may provide important information on the pathogenesis of sarcoidosis, and the display pattern implies the potential usefulness of this method as a tool for diagnosis of the disease.
结节病是一种病因不明的系统性肉芽肿性疾病。为阐明其发病机制，我们寻找了在结节病中特异性表达的已知或未知基因。采用差异显示法对18例结节病患者和8例患有各种肺部疾病的患者的支气管肺泡灌洗（BAL）细胞进行了分析。从BAL细胞中提取mRNA，并用12种锚定引物进行逆转录，理论上这些引物可覆盖所有mRNA，随后用锚定引物和一个10聚体随机引物进行聚合酶链反应（PCR）。PCR产物在聚丙烯酰胺凝胶上显示，提取结节病患者与其他患者之间强度有特征性改变的片段，进行测序，并与Genbank和EMBL DNA数据库进行比对。在结节病患者中检测到一个强度特异性增加的片段，另一个片段消失。这些片段可能来自未知基因。CD44和肿瘤坏死因子（TNF-α）cDNA序列也作为结节病中共同表达的片段被检测到。在结节病中强度特异性增加或降低的克隆片段可能为结节病的发病机制提供重要信息，并且这种显示模式表明该方法作为一种疾病诊断工具具有潜在的实用性。
