Division of Immunochemistry, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Parkallee 4a, 23845 Borstel, Germany.
Eur J Cell Biol. 2010 Jan;89(1):25-33. doi: 10.1016/j.ejcb.2009.10.010.
The lipopolysaccharide (LPS) of Pseudomonas aeruginosa has been identified to contain an inner-core structure expressing a Pseudomonas-specific epitope. This target structure is characterized by a highly phosphorylated and 7-O-carbamoyl-l-glycero-alpha-d-manno-heptopyranose (CmHep) and was found to be present in all human-pathogenic Pseudomonas species of the Palleroni (RNA)-classification I scheme. We raised and selected the monoclonal antibody S60-4-14 (mAb S60-4-14, subtype IgG1) from mice immunized with heat-killed Pseudomonas bacteria. The epitope of this mAb was found to reside in the inner-core structure of P. aeruginosa and, hence, successfully evaluated for the immunohistochemical detection of P. aeruginosa in formalin- or HOPE-fixed (Hepes-glutamic acid buffer-mediated organic solvent protection effect) and paraffin-embedded human lung tissue slices. Lung specimens, mainly from explanted lungs of cystic fibrosis (CF) patients, as well as P. aeruginosa isolates from patients suffering from CF and patients with extrapulmonar Pseudomonas infections were investigated by PCR, immunohistochemistry, and Western blot analysis with mAb S60-4-14. The results revealed an unequivocal coincidence of PCR and immunohistochemistry. Together with the Western blot results mAb S60-4-14 displays a potential diagnostic tool for the specific identification of P. aeruginosa in infected lungs of CF.
铜绿假单胞菌的脂多糖（LPS）已被确定含有表达假单胞菌特异性表位的内核心结构。该靶结构的特征是高度磷酸化和 7-O-碳氨酰-l-甘油-α-d-甘露庚糖（CmHep），并存在于所有人类致病性假单胞菌属的帕勒罗尼（RNA）-分类 I 方案中。我们从用热灭活铜绿假单胞菌免疫的小鼠中产生和选择了单克隆抗体 S60-4-14（mAb S60-4-14，亚型 IgG1）。该 mAb 的表位位于铜绿假单胞菌的内核心结构中，因此成功评估了该 mAb 在福尔马林或 HOPE 固定（Hepes-谷氨酸缓冲介导的有机溶剂保护效应）和石蜡包埋的人肺组织切片中对铜绿假单胞菌的免疫组织化学检测。通过 PCR、免疫组织化学和用 mAb S60-4-14 进行的 Western blot 分析，研究了主要来自囊性纤维化（CF）患者移植肺的肺标本，以及来自 CF 患者和肺部外假单胞菌感染患者的铜绿假单胞菌分离株。PCR 和免疫组织化学结果显示出明确的一致性。与 Western blot 结果一起，mAb S60-4-14 显示出一种潜在的诊断工具，可用于特异性鉴定 CF 感染肺中的铜绿假单胞菌。
