Immunodiagnostic value and properties of an antigen isolated with affinity chromatography using a monoclonal antibody to Trichinella spiralis

使用旋毛虫单克隆抗体通过亲和层析分离抗原的免疫诊断价值和特性

• 批准号: 02670165
• 负责人: NIIMURA Munetoshi
• 金额: $ 0.45万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02670165/
• 关键词: Trichinella spiralis; alpha -Stichosome; Monoclonal antibody; Antigen; Immunodiagnosis; Anthelmintic agent; 治療剤; 局在; αースチコソ-ム; モノクロ-ナル抗体; 防御免疫

Monoclonal antibodies were generated for the isolation of specific antigen from Trichinella spiralis(T.s). A monoclonal antibody (TS32D12) of IgG_1 class was selected according to its reactivity and specificity. We isolated 160kDa molecule (Ts-alpha 160) from alpha -stichocyte of T.s muscle larvae(Ts-L_1) with affinity chromatography using TS32D12 and examined immunodiagnostic value of Ts-alpha 160 against serum samples which had been obtained from 13 patients 33-109 days after infection. Antibodies against Ts-alpha 160 were detected with higher titers than those against Ts-crude antigen in every case. The application of Ts-alpha 160 to the ELISA eliminated false-positive reactions which were observed against Ts-crude antigen in serum samples from patients with proven helminthiasis. On the other hand, the 160kDa molecule was not detected in adult worms, embryos, nor in muscle larvae at day 14 post infection. However, it was demonstrated in alpha -stichosome of the late muscle stage lar … More vae from day 21 at least up to day 180. Antibody responses of infected BALB/c mice of T.s against the Ts- alpha 160 became detectable with low titers at week 4, followed by a rapid increased in titers at week 8. These results suggested that Ts-alpha 160 can be utilized as a unique antigen for the immunodiagnosis of human trichinosis from about 5-6 week after infection. In addition, Ts-alpha 160 was found consist of glycoprotein and E-S product derived from Ts-L_1, the epitopes recognized by the TS32D12 seemed to be lost after treatment at 100ﾟC, 2 min. The Ts-alpha 160 was elucidated by the immunogold method using electron microscope that is located in alpha granules of stichocyte of Ts-L_1. There were no significant decrease in recovery of worms among experimental groups of mice both passively transferred TS32D12 and actively transferred Ts-alpha 160. Freshly isolated T.s L_1 were incubated containing ammonium embelate in PBS, large precipitates accumulated and adhered to the oral end of the worms,and were observed to react to TS32D12 by immuno-fluorescence stain. Less

制备了旋毛虫特异性抗原的单克隆抗体。根据其反应性和特异性，筛选出IgG_1类单克隆抗体（TS 32 D12）。我们用TS 32 D_（12）亲和层析法从T_s肌幼虫（Ts-L_1）的α-stichocyte中分离出160 kDa分子（Ts-α 160），并用13例患者感染后33-109天的血清检测了Ts-α 160的免疫诊断价值。在每种情况下，检测到抗Ts-α 160的抗体的滴度均高于抗Ts-粗抗原的抗体。应用Ts-α 160的ELISA消除了假阳性反应，观察到对Ts-粗抗原的血清样品中证实蠕虫病。另一方面，在感染后第14天，在蠕虫、胚胎和肌肉幼虫中均未检测到160 kDa分子。然而，在肌肉晚期的α-stichosome中， ...更多信息 从第21天至少持续至第180天。T.s感染的BALB/c小鼠对Ts-α 160的抗体应答在第4周时变得可检测到低滴度，随后在第8周滴度迅速增加。结果提示，Ts-α 160可作为旋毛虫病免疫诊断的特异性抗原，用于旋毛虫病感染后5-6周的诊断。另外，还发现Ts-α 160由糖蛋白和来源于Ts-L_1的E-S产物组成，TS 32 D12识别的表位在100 ℃处理2 min后似乎丢失。在被动转移TS 32 D12和主动转移Ts-α 160的小鼠的实验组中，蠕虫的恢复没有显著降低。新鲜分离的T.s L_1在含恩贝酸铵的PBS中孵育后，大量沉淀物聚集并粘附于蠕虫体口端，免疫荧光染色观察到其与TS 32 D12反应。少

项目成果

Munetoshi NIIMURA and Somei KOJIMA: "Immunocytolocalization and properties of a target antigen recognized by a monoclonal antibody to Trichinella spiralis." Japanese Journal of Parasitology. 40. 253-259 (1991)

Munetoshi NIIMURA 和 Somei KOJIMA：“旋毛虫单克隆抗体识别的靶抗原的免疫细胞定位和特性。”

Munetoshi NIIMURA,Masashi KOBAYASHI and Somei KOJIMA: "Immunodiagnostic value of an α-stichocyte-derived antigen isolated by affinity chromatography for trichinosis." The Japanese Journal of Parasitology. 41. 287-293 (1992)

Munetoshi NIIMURA、Masashi KOBAYASHI 和 Somei KOJIMA：“通过亲和层析分离的 α-stichogenic 抗原对旋毛虫病的免疫诊断价值”，《日本寄生虫学杂志》41. 287-293 (1992)。

Munetoshi NIIMURA and Somei KOJIMA: "Immunocytolocalization and properties of a target antigen recognized by a monoclonal antibody to Trichinella spiralis." The Japanese Journal of Parasitology. 40. 253-259 (1991)

Munetoshi NIIMURA 和 Somei KOJIMA：“旋毛虫单克隆抗体识别的靶抗原的免疫细胞定位和特性。”

Munetoshi NIIMURA, Masashi KOBAYASHI and Somei KOJIMA: "Immunodiagnostic value of an alpha -stichocyte-derived antigen isolated by affinity chromatography for trichinosis." Japanese Journal of Parasitology. 41. 287-293 (1992)

Munetoshi NIIMURA、Masashi KOBAYASHI 和 Somei KOJIMA：“通过亲和层析分离出的 α-stichogenic 抗原对旋毛虫病的免疫诊断价值。”

Munetoshi NIIMURA,Masashi KOBAYASHI and Somei KOJIMA: "Use of an αーStichocyteーderived Antigen Isolated by Affinity Chromatography for the Immunodiagnosis of Trichinosis." The Japanese Journal of Parasitology.

Munetoshi NIIMURA、Masashi KOBAYASHI 和 Somei KOJIMA：“使用亲和色谱法分离的 α-Stichogenic 抗原用于旋毛虫病的免疫诊断。”