四个旋毛虫地理株肌幼虫虫体抗原和排泄—分泌抗原的免疫学分析  

Immunological analysis of the crude and excretory-secretory antigens of muscle larvae from four Trichinella isolates

作　　者：简莎娜 艾琳[1] 陈韶红[1] 蔡玉春[1] 卢艳[1] 陈木新[1] 吴秀萍[1] 田利光[1] 陈家旭[1] 

机构地区：[1]中国疾病预防控制中心寄生虫病预防控制所,卫生部寄生虫病原与媒介生物学重点实验室,世界卫生组织疟疾,血吸虫病和丝虫病合作中心

出　　处：《国际医学寄生虫病杂志》2015年第3期137-141,共5页International JOurnal of Medical Parasitic Diseases

基　　金：国家科技重大专项(2008ZX10004-011;2012ZX10004-220);国家科技支撑计划(2008BAI56B03)~~

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100 000、66 000、49 000、45 000、43 000、30 000、18 000、12 000和Mr 66 000、53 000、49 000、43 000、36 000、34 000、30 000、16 000。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Mr 95 000、72 000、53 000、49 000、43 000为主，排泄-分泌抗原反应条带以Mr 53 000、49 000、43 000、40 000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Mr 95 000、72 000成分和排泄-分泌抗原的Mr 40 000成分是进一步研究的重点。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 elec- trophoresis （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 TrichineUa, healthy individuals, and patients with trichinosis, schitosomiasis, clonorchiasis, echinococcosis, sparganosis, trichuriasis, paragonimiasis, eysticercosis, 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 M, 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 M, 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.

关 键 词：旋毛虫 地理株 幼虫虫体抗原 幼虫排泄-分泌抗原 

分 类 号：R383.15[医药卫生—医学寄生虫学]