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

我们进行了流行病学和血清流行病学调查的哥斯达黎加血管圆线虫感染在巴西南部，我们也进行了实验研究的诊断和免疫病理学的疾病。主要研究结果如下：1.在巴西南部，两种蛞蝓，Sarasinula linguaeformis和Phyllocaulis variegatus，都是寄生虫的重要中间宿主。我们首次发现另一种蛞蝓，Deroceraslaeve，作为中间宿主。这些蛞蝓的流行率有明显的季节性波动，而实际的流行率则与季节性波动有关。8月和12月的舌形线虫分别为1.5%和24%。采用ELISA分析法进行的血清流行病学研究表明，圣卡塔利纳新伊塔贝拉巴当地人的感染率在8月为22%，12月为35%，4月为53%。在此期间，每四个月的发病率为24%至30%。我们对人们的口头问卷调查表明，他们的感染将是 ...更多信息 因进食生蔬菜，可能被受感染的蛞蝓的粘液污染，或因使用蛞蝓作为鱼饵钓鱼等。明胶颗粒间接凝集试验。costaricensis成虫抗原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的感染，因为假阳性反应和与A.广州线虫感染。需要对纯化用于该试验的抗原进行改进。一种识别A. costaricensis成立。用单克隆抗体进行的斑点ELISA试验成功地检测到了感染的沙鼠和人类血清中的循环抗原。然而，一些改进将是必要的，以增加测试的敏感性，并消除了一个弱的交叉反应，广州管圆线虫病。为了开发一种遗传诊断方法，应用减法技术检测可能涉及感染动物血液中的蠕虫来源的DNA片段。我们已经得到了几条消减带，但还没有成功地检测到蠕虫衍生的DNA。目前正在进行差异筛选。我们使用IL-5转基因和非转基因C3 H/HeN小鼠进行的研究表明，嗜酸性粒细胞可能与宿主对疟原虫的抗性有关。costaricensis、生殖力和肠道病理学。我们对不同品系的近交系小鼠和B1 O同源系小鼠的研究表明，三个低繁殖力的品系，即，BALB/c、DBA/2和B10.D2对A. Costaricensis，而两个具有高繁殖力的菌株，即，C57 BL 6和C3 H/HeN敏感。前三株可用于建立人腹腔管圆线虫病小鼠模型。少

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。