寄生虫与医学昆虫学报2025，Vol.32Issue(1)：1-9,15,10.DOI:10.3969/j.issn.1005-0507.2025.01.001
旋毛虫核酸酶Ts87 降解小鼠中性粒细胞胞外捕获网DNA骨架的研究
TRICHINELLA SPIRALIS NUCLEASE TS87 DEGRADES THE DNA BACKBONE OF MOUSE NEUTROPHILS EXTRACELLULAR TRAP
摘要
Abstract
Objective This study aimed to investigate whether Trichinella spiralis Ts87 protein can facilitate immune evasion by degrading the mouse neutrophil extracellular traps(NETs).Methods First,we used bioinformatics tools to analyze the physicochemical properties of Ts87 and,through sequence homology alignment,confirmed its classification within the nuclease family.Molecular cloning and protein purification techniques were then employed to obtain high-purity Ts87 protein,and its nuclease activity was confirmed.Additionally,we compared the primary sequence of Ts87 with known active sites from human lysosomal DNase IIα and the structurally characterized Burkholderia thailandensis DNase II.This enabled us to predict potential active sites.Ts87 enzymatic mutants were generated using site-directed mutagenesis,and their functions were confirmed through enzymatic activity experiments.The extracellular nucleic acid content of mouse bone marrow neutrophils and the immunofluorescence staining of mouse NETs were assessed in vitro to verify whether the Ts87 protease mutants exhibited a decreased ability to degrade NETs.Finally,after immunizing mice with Ts87,calculated larvae burden and the degradation of NETs in vivo were observed by immunostaining the MPO and CitH3 expression in the intestinal NETs of the mice.Results High-purity Ts87 protein was successfully obtained and confirmed to possess nuclease activity,capable of degrading NETs in vitro.The Ts87 mutants exhibited reduced capacity in degrading NETs.In vivo experiments in mice demonstrated that the production of anti-Ts87 antibodies in immunized mice reduced the degradation of NETs,facilitating NET-mediated attack on the parasites and resulting in decreased larvae burden in the mice.Conclusions Trichinella spiralis can utilize Ts87 nuclease to evade capture by NETs,providing potential vaccine and drug targets for the prevention and treatment of trichinellosis.关键词
旋毛虫核酸酶Ts87/中性粒细胞胞外捕获网/免疫逃避Key words
Trichinella spiralis nuclease Ts87/Neutrophil extracellular traps/Immune evasion分类
医药卫生引用本文复制引用
张明明,郝瑜琬,程喻力,黄京京,贾智惠,诸欣平..旋毛虫核酸酶Ts87 降解小鼠中性粒细胞胞外捕获网DNA骨架的研究[J].寄生虫与医学昆虫学报,2025,32(1):1-9,15,10.基金项目
国家自然科学基金项目(No.81672042 （）
No.82202549) （）
