Department of Parasitology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran.
Arch Razi Inst. 2022 Feb 28;77(1):23-28. doi: 10.22092/ari.2021.352744.1578. eCollection 2022 Feb.
Scant information is available on the immunological aspect of causing linguatulosis in humans and animals. The present study aimed to analyze the content of crude somatic extracts and excretory-secretory products of nymphs to detect the immune response of sheep and immunogenic proteins of the parasite. After collecting the nymphs, somatic extracts were prepared by sonication. Excretory secretory products were prepared by the incubation of nymphs in RPMI medium at 37°C with 5% CO2. Somatic and excretory-secretory proteins were isolated using SDS-PAGE. The immunogenic properties of the resulting proteins were determined using immunoblotting and positive sera from sheep infected with visceral linguatulosis. The total content of somatic extracts and excretory-secretory products of nymphs analyzed by SDS-PAGE (12% gel) revealed two protein patterns with more than 18 and 9 strong bands, respectively. Immunoblots using sera samples of sheep infected with the parasite, somatic extracts and excretory-secretory products demonstrated 12 and 3 antigenic proteins with molecular weights mostly in the range of 24-100 kDa and an antigen more than 180 kDa. Three common immunodominant antigenic proteins with molecular weights of 38 and 57, as well as an antigen of more than 180 kDa, were detected in the somatic extracts and excretory-secretory products of nymphs in sheep with visceral linguatulosis. These antigens can be considetered prime candidates for future serodiagnosis and immunoprotective studies of the parasite.
有关 引起人类和动物类圆线虫病的免疫学方面的信息很少。本研究旨在分析幼虫的粗体提取液和排泄-分泌产物的内容物,以检测绵羊的免疫反应和寄生虫的免疫原性蛋白。收集幼虫后,通过超声处理制备体提取液。将幼虫在 RPMI 培养基中于 37°C 和 5%CO2 下孵育来制备排泄-分泌产物。使用 SDS-PAGE 分离体细胞和排泄-分泌产物中的蛋白质。使用免疫印迹法和感染内脏类圆线虫的绵羊的阳性血清来确定所得蛋白质的免疫原性。通过 SDS-PAGE(12%凝胶)分析的幼虫体提取物和排泄-分泌产物的总含量显示出两个蛋白质图谱,分别具有超过 18 和 9 条强带。使用感染寄生虫的绵羊血清样本、体细胞提取物和排泄-分泌产物进行免疫印迹显示出 12 种和 3 种抗原蛋白,分子量主要在 24-100 kDa 范围内,还有一种抗原蛋白超过 180 kDa。在患有内脏类圆线虫病的绵羊的幼虫体提取物和排泄-分泌产物中检测到三种共同的免疫优势抗原性蛋白,分子量分别为 38 和 57 kDa,以及一种超过 180 kDa 的抗原。这些抗原可以被认为是未来寄生虫血清诊断和免疫保护研究的主要候选物。