摘要
目的 分析4个旋毛虫地理株肌幼虫的虫体抗原、排泄-分泌抗原的蛋白组成和免疫学特性.方法 昆明小鼠16只,每只感染旋毛虫肌幼虫300条,45 d后人工消化法收集旋毛虫肌幼虫制备虫体抗原,幼虫体外培养获得排泄-分泌抗原.用聚丙烯酰胺凝胶电泳(polyacrylamide gel electrophoresis,SDS-PAGE)分析虫体抗原和排泄-分泌抗原的组分,并用感染小鼠血清和旋毛虫病、血吸虫病、华支睾吸虫病、棘球蚴虫病、裂头蚴病、鞭虫病、卫氏并殖吸虫病、囊尾蚴病、钩虫病患者血清与抗原进行免疫印迹反应,观察阳性反应抗原条带. 结果 经SDS-PAGE分析,4个旋毛虫地理株虫体可溶性抗原和排泄-分泌抗原均有8条主带,相对分子质量(Mr)分别为100000、66000、49000、45000、43000、30000、18000、12000和Mr66000、53000、49000、43000、36000、34000、30000、16000.免疫印迹结果显示,旋毛虫肌幼虫虫体可溶性抗原与旋毛虫病患者和感染小鼠血清的反应条带以Mr95 000、72 000、53 000、49000、43 000为主,排泄-分泌抗原反应条带以Mr53 000、49 000、43 000、40000为主.结论 各地理株不同抗原的组分和免疫学反应存在一定的差异,其中虫体可溶性抗原的M 95 000、72 000成分和排泄-分泌抗原的Mr40000成分是进一步研究的重点.
Abstract
Objective To identify the components and immunity of the crude and excretory-secretory antigens of muscle larvae of four Trichinella isolates.Methods Sixteen Kunming mice infected with 300 muscle larvae of Trichinella spiralis,respectively,and muscle larvae were collected on the 45th day after infection.The crude muscle larvae antigens(CLA) and the excretory-secretory antigens(ESA) of the muscle larvae were prepared by repeated freeze-thaw and culture procedures,respectively.Polyacrylamide gel electrophoresis (SDS-PAGE) was used to analyze the components of the CLA and ESA.Western-blot analysis was performed with sera of normal mice,mice infected with Trichinella,healthy individuals,and patients with trichinosis,schitosomiasis,clonorchiasis,echinococcosis,sparganosis,trichuriasis,paragonimiasis,cysticercosis,hookworm disease.Results By the analysis of SDS-PAGE,the CLA and ESA of the four Trichinella isolates were all presented 8 main bands with Mr 100 000,66 000,49 000,45 000,43 000,30 000,18 000,12 000 (former) and 70 000,16 000(latter) and 66 000,53 000,49 000,43 000,36 000,34 000,30 000,16 000 respectively,and 1 to 4 sub-bands with Mr 21 000～40 000 of the four isolates.Among them,5 immune reaction bands with Mr 95 000,72 000,53 000,49 000 and 43 000 of the CLA,and 4 bands with Mr 53 000,49 000,43 000,40 000 of the ESA were recognized with the sera of patient and mice infected with Trichinella spiralis respectively on Western blot.Conclusion There are some differences in components and immunological reaction of different isolates,Mr 40 000 component of CLA and Mr 95 000,72 000 components of ESA should be the focus of further study.
基金项目
国家科技重大专项(2008ZX10004-011)
国家科技重大专项(2012ZX10004-220)
国家科技支撑计划(2008BAI56B03)
NETL
NSTL
维普
万方数据