The effect of viral regulatory protein expression on gene delivery by human immunodeficiency virus type 1 vectors produced in stable packaging cell lines

被引:83
作者
Srinivasakumar, N
Chazal, N
HelgaMaria, C
Prasad, S
Hammarskjold, ML
Rekosh, D
机构
[1] UNIV VIRGINIA,MYLES H THALER CTR AIDS & HUMAN RETROVIRUS RES,CHARLOTTESVILLE,VA 22908
[2] UNIV VIRGINIA,DEPT MICROBIOL,CHARLOTTESVILLE,VA 22908
关键词
D O I
10.1128/JVI.71.8.5841-5848.1997
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
We describe the generation of stable human immunodeficiency virus type 1 (HIV-1)-packaging lines that constitutively express high levels of HIV-1 structural proteins in either a Rev-dependent or a Rev-independent fashion. These cell lines were used to assess gene transfer by using an HIV-1 vector expressing the hygromycin B resistance gene and to study the effects of Rev, Tat, and Nef on the vector titer. The Rev-independent cell lines were created by using gag-pol and env expression vectors that contain the Mason-Pfizer monkey virus (MPMV) constitutive transport element (CTE). Vector titers approaching 10(4) CFU/ml mere routinely obtained with these cell lines, as well as with the Rev-dependent cell lines, with HeLa-CD4 cells as targets. The presence of Nef and Tat in the producer cell each increased the vector titer 5- to 10-fold. Rev, on the other hand, was absolutely essential for gene transfer, unless the MPMV CTE was present in the vector. In that case, by using the Rev-independent cell lines for packaging, Rev could be completely eliminated from the system without a reduction in vector titer.
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页码:5841 / 5848
页数:8
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