Purification and characterization of protective antigen (PAG) against Leptospira interrogans

问号钩端螺旋体保护性抗原 (PAG) 的纯化和表征

• 批准号: 62570195
• 负责人: YANAGIHARA Yasutake
• 金额: $ 1.34万
• 依托单位: University of Shizuoka
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62570195/
• 关键词: Leptospira; Vaccine; Monoclonal antibody; Protective antigen; 糖脂質抗原; 活性酸素; マクロファージ; 感染防御; 感作凝集試験; Leptospira interrogons; serorar lai 017株; マクローファージ

Protective antigen (PAG), glycolipid nature substance, was extracted from Leptospira interrogans serovar lai strain 017 with chloroform-methanol-water (1:2:0.8, V/V) and was partially purified by silica gel column chromatography. In SDS-PAGE, the PAG was not detected by Coomassie brilliant blue staining, but was observed as single smear-like band by silver staining. The band corresponded to 24-30 Kd of standard protein. Since outer envelope (OE) fraction showed the same band, suggesting that the PAG is one of components of OE. Immunogenicity and protective activity of the PAG were compared with those of OE. the PAG was able to induce agglutinating antibody specifically, which is important for protection in leptospirosis, and did not induce any antibodies against cellular antigens of Leptospira except for agglutinogen in comparison with OE and whole cells. Furthermore, sera of mice immunized with the PAG exhibited opsonic activity against Leptospira by measurement of chemical luminescence derived from reactive oxygen. The PAG possessed protective activity against lethal infection of Leptospira in hamsters and seemed to be useful as component vaccine against leptospiral infection.

用氯仿-甲醇-水（1：2：0.8，V/V）从问号赖型钩端螺旋体017株中提取保护性抗原（PAG），并经硅胶柱层析部分纯化。在SDS-PAGE中，考马斯亮蓝染色未检测到PAG，而银染则观察到单一的涂片样条带。该条带对应于24-30 Kd的标准蛋白。结果表明，PAG能特异性地诱导钩体凝集抗体，与钩体凝集原和全细胞相比，PAG不能诱导抗钩体细胞抗原的抗体，但能诱导抗钩体凝集原的抗体。此外，用PAG免疫的小鼠血清通过测量来自活性氧的化学发光显示出针对钩端螺旋体的调理活性。PAG对钩端螺旋体致死性感染具有保护活性，可作为钩端螺旋体感染的组分疫苗。

项目成果

増沢俊幸: 日本細菌学雑誌. 43. 201 (1988)

Toshiyuki Masuzawa：日本细菌学杂志。43. 201 (1988)

Toshiyuki Masuzawa: "Classification of Leptospira interrogans serovar lai strain 017 by using monoclonal antibodies" Journal of Clinical Microbiology. 26. 2332-2337 (1988)

Toshiyuki Masuzawa：“使用单克隆抗体对问号钩端螺旋体血清型 lai 菌株 017 进行分类”《临床微生物学杂志》。

Toshiyuki Masuzawa: Journal of Clinical Microbiology. 26. 2332-2337 (1988)

Toshiyuki Masuzawa：临床微生物学杂志。

Toshiyuki Masuzawa: Infection and Immunity.

Toshiyuki Masuzawa：感染与免疫。

Yasutake Yanagihara: "Monoclonal antibody against surface antigen of Leptospira interrogans serovar canicola" Israel Journal of Veterinary medicine. 44. 56-58 (1988)

Yasutake Yanagihara：“针对问号钩端螺旋体血清型canicola表面抗原的单克隆抗体”以色列兽医杂志。