广州管圆线虫疫源地调查及单克隆抗12D5保护性效果的研究
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摘要
广州管圆线虫病,是一种人畜共患的寄生虫病。该病主要分布于东南亚及太平洋地区,因进食了未煮熟的含有广州管圆线虫( Angiostrongylus cantonensis)三期幼虫(L3)的螺肉而感染。许多螺,如福寿螺、皱疤坚螺、中国圆田螺、褐云玛瑙螺、东风螺,都是A. cantonensis的中间宿主。终宿主大鼠进食了受感染的螺后,L3幼虫会随着血液循环系统移动到脑部,最终到达肺部,并生长为成虫。故A. cantonensis又称为广州肺线虫。若A. cantonensis感染了人,其L3幼虫寄生在脑部可引起嗜酸性粒细胞增多性脑膜炎与脑膜脑炎,寄生在脑脊液中可引起头痛、头晕、发热、颈部强硬和面神经瘫痪等症状,严重者可致痴呆,甚至死亡。自2006年北京大规模爆发广州管圆线虫病以来,各地相关部门都加强了对广州管圆线虫病的检测与预防。因此,对A. cantonensis的疫源地调查成为当今研究的热点之一。同时,对广管圆线虫病的预防与治疗也被广泛研究。本文对深圳A. cantonensis的疫源地进行了调查,并对A. cantonensis的单克隆抗体12D5对感染小鼠的脑部嗜酸性粒细胞的抑制作用进行了研究。主要结果如下:
亚热带气候的深圳具有气温温和、多雨,土壤腐殖质含量丰富和灌木植被覆盖面积广泛等特点,而这些都为A. cantonensis的中间宿主褐云玛瑙螺、福寿螺提供了良好的生存环境,使其成为深圳主要的两栖和陆生淡水贝类软件动物,并广泛分布在常年积水的菜地、池塘、荒野和公园。通过对褐云玛瑙螺、福寿螺感染A. cantonensis幼虫情况的调查,我们发现受感染的中间宿主主要分布在深圳的西南部且其滋生地多为灌木植被,而且多分布在潮湿、腐殖质含量高的地带。同时还发现A. cantonensis幼虫对≥55g的褐云玛瑙螺的感染度显著高于<55g的褐云玛瑙螺。对A. cantonensis终宿主感染情况的调查发现,褐家鼠的感染率最高,且雌鼠的感染率高于雄鼠。
嗜酸性粒细胞是白细胞的一种,在抗寄生虫感染和Ⅰ型超敏反应中发挥重要作用。嗜酸性粒细胞在脑部的聚积和激活是A. cantonensis感染人和小鼠的一个重要特征。我们研究了A. cantonensis可溶性抗原单克隆抗体12D5对A. cantonensis引起的小鼠脑部嗜酸性粒细胞浸润的抑制作用和小鼠生存情况。根据各组小鼠的生存情况,我们发现,相对于感染组,单克隆抗体12D5能够显著地延长小鼠的生存时间,缓解广州管圆线虫病的各种病症,如竖毛、厌食、少动等。这说明,单克隆抗体12D5对于治疗广州管圆线虫病并延长生存时间可能有积极地作用。同时还对Eotaxin、IL-4和IL-5等细胞因子在脾脏中的表达水平进行了研究。结果发现,单克隆抗体12D5能降低IL-5在脾脏中的表达,从而抑制嗜酸性粒细胞的生存。虽然Eotaxin的表达量增加,但由于IL-5的表达水平降低,使嗜酸性粒细胞未能在脑部募集,并降低了嗜酸性脑膜炎的发生或减轻脑膜炎的症状。
Angiostrongyliasis is a disease in humans and animals, distributing widely in Southeast Asia and the Pacific areas, by ingesting the infective third-stage larvae (L3) of A. cantonensis that are found in raw or insufficiently cooked molluscs or crustaceans. Many kind of snails, including Pomacea canaliculata, Camacna cicatricose, Cipangopaludina chinensis, Achatina fulica and Babylomia formosae, are the intermediate hosts for A. cantonensis. When rats, definitive host of A. cantonensis, ingest infected intermediate hosts, L3 worms will move to their brains, arrive to the lung of rats lastly, and develop into adults. Thus, A. cantonensis is also called the rat lungworm. If human are infected by A. cantonensis, the third-stage larvae 3 worm (L3) can cause the eosinophils increasing cerebrospinal meningitis, if in humans central nervous system, also cause headache, dizziness, fever, neck firmness ,facial nerve paralysis, dementia and death, if in cerebrospinal fluid (CSF). Since the angiostrongyliasis occurred on a large scale in Peking in 2006, the related departments of all local governments enhanced the work about detection and prevention on angiostrongyliasis. Therefore, it is an important research area to research the epidemic focus of A. cantonensis and the prevention and cure of angiostrongyliasis. This study researched the epidemic focus of A. cantonensis in Shenzhen city and the inhibiting effect of the monoclonal antibody (mAb) 12D5 on eosinophils in brains of BABL/c mice. The results of our study are summarized as follows:
In Shenzhen, the weather is warm and rainy. There is lots of humus in soil and frutex. It offers a favourable living environment for P. canaliculata and A. fulica, which are the intermediate hosts for A. cantonensis. And, they become the main amphibious and shellfish animals in Shenzhen city and largely distribute in vegetable fields, pools, wildernesses and parks, which hold water always. The results of epidemic focus of A. cantonensis are that infected intermediate hosts distribute in the southwest of Shenzhen city, where the frutex grows thick and there are in damp, humus content high zone. And, the infectiosity of A. cantonensis L3 in A. fulica whose weight are≥55g is significantly higher than that are<55g. The results about final host of A. cantonensis are that infection rate of Rattus norvegicus is largest, and the infection rate of female is larger than that of male.
Eosinophils are a kind of leucocyte and act as an important work in resisting parasitic infection and THⅠhypersensitivity. The accumulation and activation of eosinophils in brain is an important hallmark of A. cantonensis infection in both humans and mice. We researched the inhibitory effects of the 12D5 mAb on eosinophils in brains and the survival ratio of BABL/c mice infected with A. cantonensis. According to the survival situation of each group mice, 12D5 mAb significantly extended the mouse life and also remitted symptoms of angiostrongyliasis including piloerection, asitia and fewer moving. Our results suggest that 12D5 mAb may have an important work in the cure and delaying of angiostrongyliasis. We also researched the mRNA expression levels of Eotaxin, IL-4 and IL-5 in the spleen. The results are the 12D5 mAb can reduce the expression of IL-5 and inhibits the survival of eosinophils. Although the expression of Eotaxin is increased, the reduction of IL-5 expression stops eosinophils to accumulate in brain and reduces the occurrence of eosinophilic meningitis or eased the symptoms of eoxinophilic meningitis.
亚热带气候的深圳具有气温温和、多雨,土壤腐殖质含量丰富和灌木植被覆盖面积广泛等特点,而这些都为A. cantonensis的中间宿主褐云玛瑙螺、福寿螺提供了良好的生存环境,使其成为深圳主要的两栖和陆生淡水贝类软件动物,并广泛分布在常年积水的菜地、池塘、荒野和公园。通过对褐云玛瑙螺、福寿螺感染A. cantonensis幼虫情况的调查,我们发现受感染的中间宿主主要分布在深圳的西南部且其滋生地多为灌木植被,而且多分布在潮湿、腐殖质含量高的地带。同时还发现A. cantonensis幼虫对≥55g的褐云玛瑙螺的感染度显著高于<55g的褐云玛瑙螺。对A. cantonensis终宿主感染情况的调查发现,褐家鼠的感染率最高,且雌鼠的感染率高于雄鼠。
嗜酸性粒细胞是白细胞的一种,在抗寄生虫感染和Ⅰ型超敏反应中发挥重要作用。嗜酸性粒细胞在脑部的聚积和激活是A. cantonensis感染人和小鼠的一个重要特征。我们研究了A. cantonensis可溶性抗原单克隆抗体12D5对A. cantonensis引起的小鼠脑部嗜酸性粒细胞浸润的抑制作用和小鼠生存情况。根据各组小鼠的生存情况,我们发现,相对于感染组,单克隆抗体12D5能够显著地延长小鼠的生存时间,缓解广州管圆线虫病的各种病症,如竖毛、厌食、少动等。这说明,单克隆抗体12D5对于治疗广州管圆线虫病并延长生存时间可能有积极地作用。同时还对Eotaxin、IL-4和IL-5等细胞因子在脾脏中的表达水平进行了研究。结果发现,单克隆抗体12D5能降低IL-5在脾脏中的表达,从而抑制嗜酸性粒细胞的生存。虽然Eotaxin的表达量增加,但由于IL-5的表达水平降低,使嗜酸性粒细胞未能在脑部募集,并降低了嗜酸性脑膜炎的发生或减轻脑膜炎的症状。
Angiostrongyliasis is a disease in humans and animals, distributing widely in Southeast Asia and the Pacific areas, by ingesting the infective third-stage larvae (L3) of A. cantonensis that are found in raw or insufficiently cooked molluscs or crustaceans. Many kind of snails, including Pomacea canaliculata, Camacna cicatricose, Cipangopaludina chinensis, Achatina fulica and Babylomia formosae, are the intermediate hosts for A. cantonensis. When rats, definitive host of A. cantonensis, ingest infected intermediate hosts, L3 worms will move to their brains, arrive to the lung of rats lastly, and develop into adults. Thus, A. cantonensis is also called the rat lungworm. If human are infected by A. cantonensis, the third-stage larvae 3 worm (L3) can cause the eosinophils increasing cerebrospinal meningitis, if in humans central nervous system, also cause headache, dizziness, fever, neck firmness ,facial nerve paralysis, dementia and death, if in cerebrospinal fluid (CSF). Since the angiostrongyliasis occurred on a large scale in Peking in 2006, the related departments of all local governments enhanced the work about detection and prevention on angiostrongyliasis. Therefore, it is an important research area to research the epidemic focus of A. cantonensis and the prevention and cure of angiostrongyliasis. This study researched the epidemic focus of A. cantonensis in Shenzhen city and the inhibiting effect of the monoclonal antibody (mAb) 12D5 on eosinophils in brains of BABL/c mice. The results of our study are summarized as follows:
In Shenzhen, the weather is warm and rainy. There is lots of humus in soil and frutex. It offers a favourable living environment for P. canaliculata and A. fulica, which are the intermediate hosts for A. cantonensis. And, they become the main amphibious and shellfish animals in Shenzhen city and largely distribute in vegetable fields, pools, wildernesses and parks, which hold water always. The results of epidemic focus of A. cantonensis are that infected intermediate hosts distribute in the southwest of Shenzhen city, where the frutex grows thick and there are in damp, humus content high zone. And, the infectiosity of A. cantonensis L3 in A. fulica whose weight are≥55g is significantly higher than that are<55g. The results about final host of A. cantonensis are that infection rate of Rattus norvegicus is largest, and the infection rate of female is larger than that of male.
Eosinophils are a kind of leucocyte and act as an important work in resisting parasitic infection and THⅠhypersensitivity. The accumulation and activation of eosinophils in brain is an important hallmark of A. cantonensis infection in both humans and mice. We researched the inhibitory effects of the 12D5 mAb on eosinophils in brains and the survival ratio of BABL/c mice infected with A. cantonensis. According to the survival situation of each group mice, 12D5 mAb significantly extended the mouse life and also remitted symptoms of angiostrongyliasis including piloerection, asitia and fewer moving. Our results suggest that 12D5 mAb may have an important work in the cure and delaying of angiostrongyliasis. We also researched the mRNA expression levels of Eotaxin, IL-4 and IL-5 in the spleen. The results are the 12D5 mAb can reduce the expression of IL-5 and inhibits the survival of eosinophils. Although the expression of Eotaxin is increased, the reduction of IL-5 expression stops eosinophils to accumulate in brain and reduces the occurrence of eosinophilic meningitis or eased the symptoms of eoxinophilic meningitis.
引文
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