汉滩病毒感染与整合素β3表达的相关性  

The relationship between cellular infection by Hantaan virus and expression of β3 integrin

作　　者：牟丹蕾[1] 白雪帆[1] 黄长形[1] 王英鹏[1] 潘蕾[1] 李光玉[1] 孙永涛[1] 聂青和[1] 杨为松[1] 

机构地区：[1]第四军医大学唐都医院全军感染病诊疗中心,西安710038

出　　处：《中华内科杂志》2004年第11期810-814,共5页Chinese Journal of Internal Medicine

摘　　要：目的　探讨整合素 β3表达与汉滩病毒 (HTNV)感染的关系。 方法　将人整合素 β3、αv及αⅡb真核表达载体分别及共转染至CHO细胞 ,采用间接免疫荧光测定 (IFA)、流式细胞计数(FCM)检测外源基因的表达 ;然后用HTNVA9株感染稳定转染的CHO细胞 ,应用IFA和FCM检测病毒抗原。结果　整合素 β3在共转染组细胞中高效表达 ,且目的蛋白主要定位于细胞膜上 ;β3单转染组目的蛋白虽有一定量的细胞膜表达 ,但表达量低于共转染组 (P <0 0 5 ) ;αv和αⅡb单转染组目的蛋白的表达量明显低于共转染组 (P <0 0 1) ,且定位发生变化。病毒感染率共转染细胞较高 ,CHO/αvβ3和CHO/αⅡbβ3细胞分别为 6 0 1%和 5 5 9% ;未转染 β3的细胞感染率较低 ,CHO细胞仅为 1% ;而单转染 β3细胞介于两者之间 ( 38 7% )。结论　汉滩病毒感染率与整合素 β3表达水平密切相关 ,整合素Objective To investigate the relationship between cellular entry of Hantaan virus (HTNV) and expression of β3 integrin in β3-integrin-deficient and HTNV-insusceptible China hamster ovary (CHO) cells.Methods Eukaryotic expression vector encoding human integrin β3 and eukaryotic expression vector harboring human integrin αv or αⅡb subunit cDNA were transfected into HTNV non-permissive CHO cells individually or collectively. Screening for stable transfectant clones was performed using G418 selective (culture medium. The exogenous gene expression was analyzed qualitatively and quantitatively by immunofluorescence assay (IFA) and flow cytometry (FCM). Various modified CHO cells and untransfected CHO cells were infected using HTNV A9. At various time points after infection, HTNV antigens in infected cells were detected qualitatively and quantitatively by IFA、FCM. Results Highly-effective surface expression of β3 integrin was measured in CHO/αvβ3 and CHO/αⅡbβ3, while weaker surface expression was detected in CHO/β3( P <0.05). Expression of αv or αⅡb integrin in the individually transfected group was significantly lower than in the cotransfected group( P <0.01)and the sites of localization changed. In contrast, effective surface expression was not seen when pcDNA3 was transfected alone. The infection rate of CHO/αvβ3(60.1%) and CHO/αⅡbβ3(55. 9%) cells were significantly higher than that of CHO/β3(38.7%)cells, while the infection rate of CHO/β3 was significantly higher than that of CHO/αv, CHO/pcDNA3 and CHO cells respectively. There was a close relationship between the positive percentage of HTNV A9-infected cells and expression of β3 integrin. Conclusion These results indicated that cellular entry of HTNV was related to the expression of β3 integrin.

关 键 词：整合素Β3 共转染 表达量 汉滩病毒 CHO细胞 ΑV 蛋白 高效表达 细胞膜 外源基因 

分 类 号：R373[医药卫生—病原生物学] R730[医药卫生—基础医学]