INHIBITION OF HIV-1 TAT-MEDIATED LTR TRANSACTIVATION AND HIV-1 INFECTION BY ANTI-TAT SINGLE-CHAIN INTRABODIES
INHIBITION OF HIV-1 TAT-MEDIATED LTR TRANSACTIVATION AND HIV-1 INFECTION BY ANTI-TAT SINGLE-CHAIN INTRABODIES
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DOI：
10.1002/j.1460-2075.1995.tb07140.x
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发表时间：
1995-04-03
期刊：
影响因子：
11.4
通讯作者：
MARASCO, WA
中科院分区：
文献类型：
--
作者：
MHASHILKAR, AM;BAGLEY, J;MARASCO, WA
编码抗HIV-1 Tat、外显子1或外显子2特异性单克隆抗体的重排免疫球蛋白重链和轻链可变区的基因已用于构建单链细胞内抗体“胞内抗体”，用于在哺乳动物细胞的细胞质中表达。这些抗 Tat 单链胞内抗体（抗 Tat sFv）还分别用 C 端人 C-κ 结构域进行修饰，以增加细胞质稳定性和/或 C 端 SV40 核定位信号以将新生胞内抗体引导至核区室。抗 Tat sFv 对 Tat 的 N 端激活结构域具有特异性结合活性，可阻断 Tat 介导的 HIV-1 LTR 反式激活以及哺乳动物细胞中 Tat 的细胞内运输。因此，表达抗 Tat sFv 的转化淋巴细胞能够抵抗 HIV-1 感染。因此，这些研究证明稳定表达的单链胞内抗体及其修饰形式可以有效地靶向真核细胞的细胞质和核区室中的分子。此外，这些研究表明，单独使用抗 Tat sFv 或与其他基于基因的策略组合使用可能有助于 HIV-1 感染和艾滋病的基因治疗。
Genes encoding the rearranged immunoglobulin heavy and light chain variable regions of anti-HIV-l Tat, exon 1 or exon 2 specific monoclonal antibodies have been used to construct single chain intracellular antibodies 'intrabodies' for expression in the cytoplasm of mammalian cells. These anti-Tat single chain intrabodies (anti-Tat sFvs) are additionally modified with a C-terminal human C-kappa domain to increase cytoplasmic stability and/or the C-terminal SV40 nuclear localization signal to direct the nascent intrabody to the nuclear compartment, respectively. The anti-Tat sFvs with specific binding activity against the N-terminal activation domain of Tat, block Tat-mediated transactivation of HIV-1 LTR as well as intracellular trafficking of Tat in mammalian cells. As a result, the transformed lymphocytes expressing anti-Tat sFvs are resistant to HIV-1 infection. Thus, these studies demonstrate that stably expressed single chain intrabodies and their modified forms can effectively target molecules in the cytoplasm and nuclear compartments of eukaryotic cells. Furthermore, these studies suggest that anti-Tat sFvs used either alone or in combination with other genetically based strategies may be useful for the gene therapy of HIV-1 infection and AIDS.