细粒棘球绦虫表面抗原热休克蛋白抗体在免疫组化中的应用  

Immunohistochemistry application of antibodies to Echinococcus granulosus surface antigen heat shock protein

作　　者：路鹏霏 田梦潇 齐洪志[1] 包永星[1] 毛睿[1] LU Peng-fei;TIAN Meng-xiao;QI Hong-zhi;BAO Yong-xing;MAO Rui(Tumor Center of the First affiliated Hospital of Xinjiang Medical University,Urumqi 830011,China;College of Basic Medicine,Xinjiang Medical University,Urumqi 830011,China)

机构地区：[1]新疆医科大学第一附属医院肿瘤中心,乌鲁木齐830011 [2]新疆医科大学基础医学院,乌鲁木齐830011

出　　处：《中国人兽共患病学报》2023年第5期440-446,共7页Chinese Journal of Zoonoses

基　　金：国家自然科学基金项目(No.81860360,No.81860556);省部共建中亚高发病成因与防治国家重点实验室课题(No.SKL-HIDCA-2020-27)联合资助。

摘　　要：目的用生物信息工具分析细粒棘球绦虫表面抗原热休克蛋白70(EgHSP70)的理化性质、结构域等信息,并制备多克隆抗体进行人、羊、小鼠肝包虫病的免疫组化研究。方法EgHSP70氨基酸序列经NCBI数据库中下载,用ProtParam(性质)、SignalP-5(信号肽)、SOSUI(亚细胞定位)、ProtScale、SOSUI、DNASTAR(亲疏水性)、SOMPA(二级结构)、Swissmodel(三级结构)、TMHMM(跨膜区域)工具分析。制备多克隆蛋白抗体,分别进行人、绵羊、C57BL/6J小鼠肝包虫的免疫组化染色。结果EgHSP70是由258个氨基酸序列组成,分子式为C 1248 H 2024 N 354 O 392 S 2,不稳定指数为42.00,为亲水蛋白,无信号肽,无跨膜区,且定位于细胞质中。二级结构中,α螺旋占47.29%,β折叠占16.28%,β转角占8.53%,无规则卷曲占27.91%。免疫组化结果显示,EgHSP70多克隆抗体能够在细粒棘球绦虫的生发层(包囊)细胞中高表达,在羊、C57小鼠正常肝组织中少量表达,在人肝脏组织中不表达。结论免疫组化检测EgHSP70的多克隆抗体在不同的物种组织中,能识别生发层细胞及细粒棘球蚴虫体细胞。This study was aimed at analyzing the physicochemical properties and domains of Echinococcus granulosus antigen heat shock protein 70(EgHSP70)by using bioinformatics tools,and to prepare polyclonal antibodies for immunohistochemical analysis of liver echinococcosis in humans,sheep and mice.The amino acid sequences of EgHSP70 were downloaded from the NCBI database and analyzed with ProtParam(property),SignalP-5(signal peptide),SOSUI(subcellular localization),ProtScale,SOSUI,DNASTAR(hydrophobicity),SOMPA(secondary structure),Swissmodel(tertiary structure)and TMHMM(transmembrane region).Polyclonal protein antibodies were prepared and used for immunohistochemical staining of hepatic echinococcosis in humans,sheep and C57BL/6J mice.EgHSP70 was composed of 258 amino acids and had a molecular formula of C 1248 H 2024 N 354 O 392 S 2;its instability index was 42.00.It was a hydrophilic protein with no signal peptide or transmembrane region,and it localized to the cytoplasm.The secondary structure comprised 47.29%αhelices,16.28%βfolds,8.53%βcorners and 27.91%irregular crimps.Immunohistochemis try indicated that EgHSP70 polyclonal antibody was expressed at high levels in the germinal layer(cyst)cells of Echinococcus granulosus,and low levels in the livers of sheep and C57 mice,but was not expressed in human liver.The physical and chemical properties and structure of EgHSP70 were predicted by bioinformatics,and the germinal layer cells and somatic cells of Echinococcus granulosus were identified in different species by immunohistochemistry.

关 键 词：细粒棘球绦虫 EgHSP70 生物信息学 免疫组化 

分 类 号：R383.3[医药卫生—医学寄生虫学]