Levin R, Mhashilkar A M, Dorfman T, Bukovsky A, Zani C, Bagley J, Hinkula J, Niedrig M, Albert J, Wahren B, Göttlinger H G, Marasco W A
Division of Human Retrovirology, Dana-Farber Cancer Institute, Boston, MA 02115, USA.
Mol Med. 1997 Feb;3(2):96-110.
The HIV-1 matrix (MA) protein, p17, contains two subcellular localization signals that facilitate both nuclear import of the viral preintegration complex early during infection and virus particle assembly late in infection. The dual role of MA in both the afferent and efferent arms of the HIV-1 life cycle makes it an important target for intracellular immunization-based gene therapy strategies.
Here we report, using a new bicistronic vector, that an intracellular Fab antibody, or Fab intrabody, directed against a carboxy-terminal epitope of MA from the Clade B HIV-1 genotype, can inhibit HIV-1 infection when expressed in the cytoplasm of actively dividing CD4+ T cells.
Marked inhibition of proviral gene expression occurred when single-round HIV-1 CAT virus was used for infections. In challenge experiments using both laboratory strains and syncytium-inducing primary isolates of HIV-1, a substantial reduction in the infectivity of virions released from the cells was also observed.
This novel strategy of simultaneously blocking early and late events of the HIV-1 life cycle may prove useful in clinical gene therapy approaches for the treatment of HIV-1 infection and AIDS, particularly when combined with genetic or pharmacologic-based strategies that inhibit other HIV-1 target molecules simultaneously.
HIV-1基质（MA）蛋白p17含有两个亚细胞定位信号，在感染早期促进病毒前整合复合物的核输入，在感染后期促进病毒颗粒组装。MA在HIV-1生命周期的传入和传出环节中的双重作用使其成为基于细胞内免疫的基因治疗策略的重要靶点。
在此我们报告，使用一种新的双顺反子载体，一种针对B亚型HIV-1基因型MA羧基末端表位的细胞内Fab抗体（或Fab胞内抗体），当在活跃分裂的CD4+T细胞质中表达时，可抑制HIV-1感染。
当使用单轮HIV-1 CAT病毒进行感染时，前病毒基因表达受到显著抑制。在使用HIV-1实验室毒株和诱导合胞体的原代分离株进行的挑战实验中，还观察到从细胞释放的病毒颗粒的感染性大幅降低。
这种同时阻断HIV-1生命周期早期和晚期事件的新策略可能在治疗HIV-1感染和艾滋病的临床基因治疗方法中证明是有用的，特别是与同时抑制其他HIV-1靶分子的基于基因或药理学的策略联合使用时。
