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

我们在巴西南部进行了血管造成的肋骨感染的流行病学和血清ePidemiologicy Surveys，我们还对疾病的诊断和免疫病理学进行了实验研究。获得的结果如下：1。在巴西南部，slugs，sarasinula linguaeformis和phyllocaulis variegatus都是寄生虫的重要中间宿主。我们首先透露，另一个Slug Deroceras Laeve是中间主机。这些sl的患病率显示出明显的季节性波动，而八月份的语言链球菌的实际患病率为1.5％，12月为24％。使用ELISA测定法的一项血清ePIDEMIologicy研究表明，圣卡塔利纳州诺瓦伊塔巴的当地人的患病率在8月份为22％，12月35％，四月为53％。在此期间，每四个月的事件率从24％到30％不等。我们对人们的口头问卷提出，他们的感染将是……更多是由于吃生蔬菜而引起的，可能被感染的sl污染物污染，或者将sl肉用作钓鱼等诱饵等。2。发现明胶颗粒间接凝集测试对粗曲霉的成年蠕虫抗原可用于鼠感染的免疫诊断，但对于其他实验动物和人类的感染而言，由于假阳性反应和对A. antonensis感染的交叉反应较弱，因此对其他实验性动物和人类的感染没有用。净化抗原进行测试将需要改进。单克隆抗体确定了成年蠕虫的极端和秘密产品中的162 kD抗原。用单克隆抗体成功地检测到感染的jird和人类血清中的循环抗原。但是，对于提高测试的灵敏度以及消除对血管造成的较弱的血管造血管的敏感性将是必要的一些改进。为了开发一种遗传诊断方法，应用了一种减法技术来检测可能参与感染动物血液的蠕虫衍生的DNA碎片。我们已经获得了几个减去带，但尚未成功检测到蠕虫来源的DNA。现在正在进行差异筛查3。我们使用IL-5转基因和非转基因C3H/HEN小鼠的研究表明，嗜酸性粒细胞可能与宿主对肋骨曲霉，粪便和肠道病理学的抗性有关。我们对各种杂交小鼠和B1O相似菌株的研究表明，三种繁殖力较低的菌株，即BALB/C，DBA/2和B10.D2，对A. costaricensis具有抵抗力，而两种具有高繁殖力的菌株，即高繁殖力，即C57BL6和C57BL6和C3H/C3H/HER HER，均为均匀。前三种菌株将有助于在小鼠中开发人类的腹部血管造血模型。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.

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。

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。