Ono E, Takase H, Naiki M, Yanagawa R
Department of Hygiene and Microbiology, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Japan.
J Gen Microbiol. 1987 May;133(5):1329-36. doi: 10.1099/00221287-133-5-1329.
A glycolipid antigen possessing a serovar-specific antigenic determinant of Leptospira interrogans serovar canicola was purified from a chloroform/methanol extract of the organism. The purification procedures included silicic acid column chromatography and preparative thin-layer chromatography (TLC). Antigenic activity was detected by a TLC-enzyme immunostaining technique using monoclonal antibody CT3, which specifically agglutinates serovar canicola and only weakly serovar sumneri but no other serovars of Leptospira. The purified glycolipid reacted with CT3 antibody, indicating that the glycolipid possessed a serovar-specific antigenic determinant. Infrared spectrum and proton nuclear magnetic resonance analyses showed that the glycolipid contained sugar and lipid moieties, which possessed amide linkages and an acetyl group. Gas-liquid chromatography-mass spectrometry analysis showed that the glycolipid contained two unknown sugars, one of which (unknown sugar II) appeared to be associated with the antigenic determinant specific for canicola. The serovar-specific antigenic determinant was destroyed by mild alkali treatment of the glycolipid. These findings suggested that the antigenic determinant was an alkali-labile moiety which may be related to the unknown sugar II.
从问号钩端螺旋体犬型血清型的氯仿/甲醇提取物中纯化出一种具有犬型血清型特异性抗原决定簇的糖脂抗原。纯化程序包括硅酸柱色谱和制备型薄层色谱（TLC）。使用单克隆抗体CT3通过TLC酶免疫染色技术检测抗原活性，该抗体特异性凝集犬型血清型，对苏门答腊血清型凝集作用较弱，对其他钩端螺旋体血清型无凝集作用。纯化的糖脂与CT3抗体反应，表明该糖脂具有血清型特异性抗原决定簇。红外光谱和质子核磁共振分析表明，该糖脂含有糖和脂质部分，具有酰胺键和一个乙酰基。气液色谱-质谱分析表明，该糖脂含有两种未知糖，其中一种（未知糖II）似乎与犬型特异性抗原决定簇有关。通过对糖脂进行温和碱处理，血清型特异性抗原决定簇被破坏。这些发现表明，抗原决定簇是一种对碱不稳定的部分，可能与未知糖II有关。
