猪细小病毒样颗粒在树突状细胞中定位的研究  

Location study of recombinant parvovirus-like particles in dendritic cells

作　　者：申小强 王超[2] 左玉柱[1] 范慧霞[3] 范京惠[1] 顾文源[1] 邸晶美 

机构地区：[1]河北农业大学动物医学院,河北保定071001 [2]泰安出入境检验检疫局,山东泰安271000 [3]河北农业大学信息科学与技术学院,河北保定071001

出　　处：《河北农业大学学报》2014年第6期95-100,共6页Journal of Hebei Agricultural University

基　　金：国家自然基金资助项目(31101847)

摘　　要：为了探明猪细小病毒样颗粒(PPV-VLPs-E290)在树突状细胞(DC)中的定位情况,首先通过磁性筛选的方法从猪的脾脏分离DC,将病毒样颗粒用荧光素FITC标记后,体外与DC在37℃下作用,应用流式细胞仪检测在体外DC对PPV-VLPs的捕获情况。体内捕获情况则是将FITC标记的PPV-VLPs免疫仔猪后,分离DC进行检测。捕获PPV-VLPs的DC用皂苷透化后,分别与内吞体的表面标志分子Rab5、Rab7、Rab11、Lamp2单克隆抗体及PE标记的PPV-VP2抗体反应,通过共聚焦显微镜检测PPV-VLPs在DC中的定位情况。结果显示,DC在体内和体外均能有效捕获PPV-VLPs,PPV-VLPs可与晚期内吞体相关蛋白分子Lamp2及Rab7的单抗共定位,而与Rab5、Rab11不共定位,表明PPV-VLPs被DC捕获后,定位于DC的晚期内吞体。Recombinant parvovirus-like particles （PPV-VLPs） are particulate exogenous Ags that induce strong CTL response. In order to decipher the mechanisms responsible for CTL activation by such exogenous Ag, we analyzed in vivo and in vitro the capture of PPV-VLPs by dendritic cells （DCs） and the location of PPV-VLPs in DCs. The results analyzed by FAC- Star cytometer showed that PPV-VLPs are very efficiently captured by DCs. And the confocal microscopy revealed that the captured PPV-VLPs colocalized with Lamp2 and rab7 labeling, which are associated with late endosomes. Whereas no colocalization of PPV-VLPs was ob- served with rah5 and rab11, a crucial element in the control of traffic through the recycling en-dosome and a crucial element associated with early endosomes. Therefore, after endocytosis, PPV-VLPs reach late endosomes.

关 键 词：病毒样颗粒 树突状细胞 定位 

分 类 号：S852.65[农业科学—基础兽医学]