Department of Parasitology, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
Department of Parasitology, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan; Department of Medical Laboratory Science and Biotechnology, College of Health Sciences, Kaohsiung Medical University, Kaohsiung, Taiwan.
J Microbiol Immunol Infect. 2020 Aug;53(4):604-611. doi: 10.1016/j.jmii.2018.09.010. Epub 2018 Oct 18.
BACKGROUND/PURPOSE: Eosinophils are recruited to the brain of mice after infection with Angiostrongylus canonensis. Several factors produced by infected mice are well known playing the role to chemoattract eosinophils from the blood into the brain. The purpose of this study is to investigate whether Angiostronylus cantonensis young-adult worms (AcYA) have components which have eosinophilic chemotactic activity.
Eosinophil chemotactic activity of AcYA was tested by Boyden blind-well chamber technique. The components of AcYA were analysed by SDS-PAGE and Mass spectrometry. Furthermore, galectin-9 in the cerebrospinal fluid (CSF) of infected mice and galectin-9-like in AcYA were measured by ELISA technic and also were recognized by western blot analysis respectively.
Excretory-secretory products of AcYA did not show eosinophil chemotactic activity. However, the extracts of AcYA showed protein concentration-dependent eosinophil chemotactic activity and reached the peak at the 24 μg/ml. The eosinophil chemotactic activity was significantly reduced by lactose. The components of AcYA at molecular weights of approximatively 15 kDa and 35 kDa showed several galectins component in Mass spectrometric analysis. Furthermore, galectin-9-like in AcYA was recognized by ELISA and western blot analysis. In parallel with increase of galectin-9 in the CSF, eosinophils were also significantly increased in mouse after infected with A. cantonensis.
Galectin-9-like in AcYA and galectin-9 in mouse CSF were confirmed demonstrating eosinophil chemotactic activity both in vitro study and in the infection of mouse in this study.
背景/目的：旋尾线虫感染小鼠后，嗜酸性粒细胞被招募到小鼠的大脑中。已知感染小鼠产生的几种因子在趋化嗜酸性粒细胞从血液进入大脑方面发挥作用。本研究的目的是研究旋尾线虫成虫（AcYA）是否具有嗜酸性粒细胞趋化活性的成分。
通过 Boyden 盲孔室技术测试 AcYA 的嗜酸性粒细胞趋化活性。通过 SDS-PAGE 和质谱分析 AcYA 的成分。此外，通过 ELISA 技术分别测量感染小鼠脑脊液中的半乳糖凝集素-9 和 AcYA 中的半乳糖凝集素-9 样蛋白，并通过 Western blot 分析进行鉴定。
AcYA 的排泄分泌产物没有显示出嗜酸性粒细胞趋化活性。然而，AcYA 的提取物显示出蛋白浓度依赖性的嗜酸性粒细胞趋化活性，在 24μg/ml 时达到峰值。乳糖显著降低了嗜酸性粒细胞趋化活性。AcYA 的分子量约为 15 kDa 和 35 kDa 的成分在质谱分析中显示出几种半乳糖凝集素成分。此外，ELISA 和 Western blot 分析鉴定 AcYA 中存在半乳糖凝集素-9 样蛋白。与 CSF 中半乳糖凝集素-9 的增加平行的是，感染旋尾线虫后小鼠中的嗜酸性粒细胞也显著增加。
本研究在体外研究和小鼠感染实验中证实，AcYA 中的半乳糖凝集素-9 样蛋白和小鼠 CSF 中的半乳糖凝集素-9 都具有嗜酸性粒细胞趋化活性。
