2型腺伴随病毒生物安全性的初步形态学观察  

A PRELIMINARY MORPHOLOGICAL STUDY ON THE BIOLOGICAL SAFETY OF WILD TYPE 2 ADENO-ASSOCIATED VIRUS

作　　者：张桐[1] 王文亮[1] 候云德[2] 杨新科[2] 王伯云[1] 

机构地区：[1]西安第四军医大学病理学教研室,西安710033 [2]中国预防医学科学院病毒学研究所病毒基因工程国家重点实验室

出　　处：《西安医科大学学报》1998年第2期164-167,共4页Journal of Xi'an Medical University(Chinese)

基　　金：国家863基因治疗重大关键技术项目!863-BH03-05-02

摘　　要：用野生型AAV－2感染宿主293细胞后对细胞进行了形态学观察，并采用AAVCap基因探针检测了感染细胞标本中的野生型AAVDNA；采用携带β-半乳糖苷酶基因，而不含AAVrep基因及其表达产物的重组AAV作对照，感染293细胞后检测细胞内β-半乳糖苷酶表达产物，同时对细胞作形态学观察。结果显示野生型AAV－2感染宿主细胞后24～48h，多数细胞胞体收缩、变形，部分细胞死亡、脱落，用Cap基因探针从病变的293细胞基因组DNA中检测到了野生AAVDNA；而被重组AAV感染的293细胞表达了β-半乳糖苷酶，而未出现细胞病变，本结果提示野生型AAV对敏感宿主细胞可表现出毒性作用，其原因可能与rep产物的作用有关。A wild type of adeno-associatedvirus type 2 (AAV-2) was .used to infect host 293cells. Morphological changes of the cells were observed and cap gene probe of AAV was used toidenify AAV DNA in the infected cells with dot blotnucleic acid hybridization. A rep-negative recombinant AAV bearing LacZ was used as a control. After293 cells were infected with the rAAV, LacZ products were detected and the morphological observation was done as well. The results showed: 24 ~ 48hours after infection of 293 cells with the wild typeAAV-2, most of the cells shrunk or deformed inshape,some of the cells died. AAV DNA was identified in the infected 293 cells,whilst the cells infectedwith the rAAV expressed LacZ gene without cellular pathological effects. The results were suggestivethat wild type of AAV-2 is toxic to its sensitive hostcells.

关 键 词：腺伴随病毒 形态学 293细胞 安全性 载体 

分 类 号：Q782[生物学—分子生物学]