Selvam M P, Buck S M, Blay R A, Mayner R E, Mied P A, Epstein J S
Center for Biologics, Evaluation and Research, US Food and Drug Administration, Rockville, MD 20852, USA.
Antiviral Res. 1996 Dec;33(1):11-20. doi: 10.1016/s0166-3542(96)00993-x.
The sequence-specific suppression of HIV-1 replication using CD4 monoclonal-antibody-targeted liposomes, containing Rev antisense phosphorothioate oligonucleotides is described. Liposomes were prepared by encapsulating the 20-mer antisense DNA sequence of the rev HIV-1 regulatory gene, in the form of a phosphorothioate oligonucleotide. Specific targeting was accomplished by conjugating anti-CD4 mouse monoclonal antibody to the surface of the liposomes. HIV-1-infected H9 cells as well as peripheral blood T-lymphocytes were incubated with the immunoliposomes of antisense found to have potential antiviral effect. HIV-1 replication was reduced by 85% in antisense immunoliposome-treated H9 cells and peripheral blood lymphocytes, whereas the inhibition of HIV-1 replication was not observed using either empty immunoliposomes or immunoliposomes containing scrambled Rev phosphorothioate oligonucleotide sequences. The antiviral activity of both the free and the encapsulated oligonucleotides were assessed by p24, reverse transcriptase (RT) assays and polymerase chain reaction (PCR) analysis. Liposome preparations demonstrated minimal toxicity in H9 as well as in peripheral blood lymphocyte cell culture experiments. These in vitro culture results demonstrate the potential efficacy of immunoliposomes to inhibit HIV replication.
本文描述了使用含有Rev反义硫代磷酸酯寡核苷酸的、靶向CD4单克隆抗体的脂质体对HIV-1复制进行序列特异性抑制的方法。脂质体通过将HIV-1调节基因rev的20聚体反义DNA序列以硫代磷酸酯寡核苷酸的形式包封来制备。通过将抗CD4小鼠单克隆抗体缀合到脂质体表面来实现特异性靶向。将HIV-1感染的H9细胞以及外周血T淋巴细胞与已发现具有潜在抗病毒作用的反义免疫脂质体一起孵育。在反义免疫脂质体处理的H9细胞和外周血淋巴细胞中，HIV-1复制减少了85%，而使用空免疫脂质体或含有乱序Rev硫代磷酸酯寡核苷酸序列的免疫脂质体均未观察到对HIV-1复制的抑制。通过p24、逆转录酶（RT）测定和聚合酶链反应（PCR）分析评估了游离和包封寡核苷酸的抗病毒活性。脂质体制剂在H9以及外周血淋巴细胞细胞培养实验中显示出最小的毒性。这些体外培养结果证明了免疫脂质体抑制HIV复制的潜在功效。
