Stellrecht K A, Sperber K, Pogo B G
Department of Neoplastic Diseases, Mount Sinai School of Medicine, New York, New York 10029-6574.
J Virol. 1992 Apr;66(4):2051-6. doi: 10.1128/JVI.66.4.2051-2056.1992.
A variety of DNA viruses are known to activate gene expression directed by the long terminal repeat (LTR) of human immunodeficiency virus type 1 (HIV-1). In light of the proposed use of recombinant vaccinia virus for HIV-1 vaccines, evaluation of the role of vaccinia virus in HIV-1 activation is warranted. To investigate whether vaccinia virus induces HIV LTR-directed gene expression, transient expression assays in Jurkat cells persistently infected with vaccinia virus (Jvac) using plasmid DNA containing the LTR linked to the bacterial chloramphenicol acetyltransferase (CAT) gene were performed. CAT activity in Jvac cells was always recorded, although the level appears to fluctuate independently of virus titers. Dual intracytoplasmic staining and fluorescence-activated cell sorter analysis showed that CAT activity was expressed in the infected cells. CAT expression was not due to plasmid replication, since plasmid DNA extracted from Jvac cells 48 h after transfection was restricted only by enzymes which recognize methylated sequences, indicating a prokaryotic source for the DNA. These findings suggest that a factor(s) present in vaccinia virus-infected cells is capable of activating the LTR of HIV-1.
已知多种DNA病毒可激活由1型人类免疫缺陷病毒（HIV-1）的长末端重复序列（LTR）指导的基因表达。鉴于提议将重组痘苗病毒用于HIV-1疫苗，有必要评估痘苗病毒在HIV-1激活中的作用。为了研究痘苗病毒是否诱导HIV LTR指导的基因表达，使用含有与细菌氯霉素乙酰转移酶（CAT）基因相连的LTR的质粒DNA，对持续感染痘苗病毒的Jurkat细胞（Jvac）进行了瞬时表达分析。Jvac细胞中的CAT活性总是被记录到，尽管其水平似乎独立于病毒滴度而波动。双重胞质内染色和荧光激活细胞分选分析表明，CAT活性在感染细胞中表达。CAT表达不是由于质粒复制，因为转染后48小时从Jvac细胞中提取的质粒DNA仅被识别甲基化序列的酶限制，表明该DNA来源于原核生物。这些发现表明，痘苗病毒感染细胞中存在的一种或多种因子能够激活HIV-1的LTR。
