Studies on the epidemiology, diagnosis and immunopathology of angiostrongyliasis costaricensis

哥斯达黎加管圆线虫病的流行病学、诊断及免疫病理学研究

• 批准号: 12576006
• 负责人: YOSHIMURA Kentaro
• 金额: $ 4.99万
• 依托单位: AKITA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12576006/
• 关键词: Angiostrongylus costaricensis; Brazil; slug; ELISA; monoclonal antibody; circulating antigen; IL-5 transgenic mouse; genetic diagnosis; B10 congenic mice; DNA診断; Sarasinula linguaeformis; IL-5 transgenic mice; Gelatin Particle Agglutination Te

We performed epidemiological and seroepidemiological surveys of Angiostrongylus costaricensis infections in southern Brazil, and also we carried out experimental studies on the diagnosis and immunopathology of the disease. The results obtained are as follows:1. In southern Brazil, both slugs, Sarasinula linguaeformis and Phyllocaulis variegatus, are an important intermediate host for the parasite. We firstly revealed that another slug, Deroceras laeve, serves as an intermediate host. The prevalence rate of these slugs showed a distinct seasonal fluctuation and the actual prevalence rate of S. linguaeformis was 1.5% in August and 24% in December. A seroepidemiological study using ELISA assays indicated that the prevalence rate of local people in Nova Itaberaba, Santa Catalina, was 22% in August, 35% in December and 53% in April. The incidence rate every four months during this period ranged from 24 to 30%. Our verbal questionnaires to the people suggested that their infections would be … More caused by eating raw vegetables, possibly contaminated with mucus from infected slugs, or by using slugs as a bait for fishing and so on.2. Gelatin particle indirect agglutination test with crude A. costaricensis adult worm antigen was found to be useful for the immunodiagnosis of murine infections but not for the infections in other experimental animals and humans, since false positive reactions and a weak cross-reaction to A. cantonensis infection were noted. An improvement will be necessary for purifying antigens for the test. A monoclonal antibody recognizing a 162 kD antigen in excretory and secretory products by adult worms ofA. costaricensis was established. A dot-ELISA test with the monoclonal antibody successfully detected a circulating antigen in the sera from infected jirds and humans. However, some improvements will be necessary for increasing the sensitivity of the test and also for eliminating a weak cross-reaction to angiostrongyliasis cantonensis. To develop a genetic diagnostic method, a subtraction technique was applied to detect worm-derived DNA fragment(s) that would be possibly involved in the blood of infected animals. We have obtained several subtracted bands but not succeeded in detecting the worm-derived DNA yet. Differential screening was now in progress.3. Our studies using IL-5 transgenic and non-transgenic C3H/HeN mice demonstrated that eosinophils were possibly associated with host resistance to A. costaricensis, fecundity and intestinal pathology. Our studies with various strains of inbred mice and B1O congenic strains revealed that three strains with a low fecundity, i.e., BALB/c, DBA/2 and B10.D2, were resistant to A. costaricensis whereas two strains with a high fecundity, i.e., C57BL6 and C3H/HeN, were susceptible. The former three strains will be useful for developing a human model of abdominal angiostrongyliasis in the mouse. Less

我们对巴西南部哥斯达黎加管圆线虫感染进行了流行病学和血清流行病学调查，并对该疾病的诊断和免疫病理学进行了实验研究。得到的结果如下： 1．在巴西南部，两种蛞蝓——Sarasinula linguaeformis和Phyllocaulis variegatus，都是这种寄生虫的重要中间宿主。我们首先揭示了另一种蛞蝓，Deroceras laeve，作为中间宿主。蛞蝓的流行率呈现明显的季节性波动，8月份舌形蛞蝓的实际流行率为1.5%，12月份为24%。采用ELISA检测的血清流行病学研究表明，圣卡塔利娜州新伊塔贝拉巴当地居民8月份的患病率为22%，12月份为35%，4月份为53%。在此期间每四个月发病率在24%至30%之间。我们对人们进行的口头调查表明，他们的感染可能是由于吃生蔬菜引起的，可能是被受感染的蛞蝓的粘液污染，或者使用蛞蝓作为钓鱼的诱饵等引起的。2。发现用粗制哥斯达黎加线虫成虫抗原进行明胶颗粒间接凝集试验可用于小鼠感染的免疫诊断，但不适用于其他实验动物和人类的感染，因为注意到假阳性反应和与广州线虫感染的弱交叉反应。为了纯化测试用抗原，需要进行改进。一种单克隆抗体，可识别 A 成虫排泄物和分泌产物中的 162 kD 抗原。哥斯达黎加成立。使用单克隆抗体进行的斑点 ELISA 测试成功地检测到了受感染沙鼠和人类血清中的循环抗原。然而，为了提高测试的灵敏度以及消除与广州管圆线虫病的微弱交叉反应，需要进行一些改进。为了开发遗传诊断方法，应用了减法技术来检测可能存在于受感染动物血液中的蠕虫来源的 DNA 片段。我们已经获得了几个消减条带，但尚未成功检测到蠕虫来源的 DNA。目前正在进行差异筛选。3．我们使用 IL-5 转基因和非转基因 C3H/HeN 小鼠进行的研究表明，嗜酸性粒细胞可能与宿主对哥斯达黎加放线菌的抵抗力、繁殖力和肠道病理有关。我们对各种近交系小鼠品系和 B1O 同源品系的研究表明，三个低繁殖力品系，即 BALB/c、DBA/2 和 B10.D2，对哥斯达黎加 A. costaricensis 具有抗性，而两个高繁殖力品系，即 C57BL6 和 C3H/HeN，则敏感。前三种菌株可用于开发小鼠腹部血管圆线虫病的人类模型。较少的

项目成果

Yoshimura, K.: "Topley & Wilson's Microbiology and Microbial Infections. Vol.5 Parasitology Edited by Cox, F.E.C. et al.(分担執筆)"Arnold, London, UK(in press).

Yoshimura, K.：“Topley & Wilsons Microbiology and Microbial Infections. Vol.5 Parasitology Edited by Cox, F.E.C. et al. (contributor)” Arnold，伦敦，英国（出版中）。

Yoshimura, K.: "Topley & Wilson's Microbiology and Microbial Infections. Vol.5 Parasitology. Edited by Cox, F. E. C. et al.(分担執筆)"Arnold, London, UK (in press).

Yoshimura, K.：“Topley & Wilsons Microbiology and Microbial Infections. Vol.5 Parasitology. Edited by Cox, F. E. C. et al. (contributor)” Arnold，伦敦，英国（出版中）。

Sugaya, H. et al.: "Eosinophils in the cerebrospinal fluid of mice infected with Angiostrongylus cantonensis are resistant to apoptosis"Int. J. Parasitol. 31. 1649-1658 (2001)

Sugaya, H. 等人：“感染广州管圆线虫的小鼠脑脊液中的嗜酸性粒细胞对细胞凋亡具有抵抗力”Int.

Sugaya, H. et al.: "Eosinophils in cerebrospinal fluid of mice infected with Angiostrongylus cantonensis are resistant to apoptosis"Int.J.Parasitol. 31(14). 1649-1658 (2001)

Sugaya, H. 等人：“感染广州管圆线虫的小鼠脑脊液中的嗜酸性粒细胞对细胞凋亡具有抵抗力”Int.J.Parasitol。

Maurer R.L. et al.: "Natural infection of Deroceras laeve (Mollusca,Gastropoda) with metastrongylid larvae in a transmission focus abdominal angiostrongyliasis"Rev. Inst. Med. Trop. Sao Paulo. 44(1). 43-44 (2002)

Maurer R.L. 等人：“腹部血管圆线虫病传播灶中后圆线虫幼虫对 Deroceras laeve（软体动物、腹足纲）的自然感染”Rev。