Joshi S, Van Brunschot A, Asad S, van der Elst I, Read S E, Bernstein A
Department of Microbiology, University of Toronto, Ontario, Canada.
J Virol. 1991 Oct;65(10):5524-30. doi: 10.1128/JVI.65.10.5524-5530.1991.
Human immunodeficiency virus type 1 (HIV-1) primarily infects CD4+ lymphocytes and macrophages and causes AIDS in humans. Retroviral vectors allowing neomycin phosphotransferase (npt) gene expression were engineered to express 5' sequences of HIV-1 RNA in the antisense or sense orientation and used to transform the human CD4+ lymphocyte-derived MT4 cell line. Cells expressing antisense or sense RNA to the HIV-1 tat mRNA leader sequence, as part of the 3' untranslated region of the npt mRNA, remained sensitive to HIV-1 infection. In contrast, resistance to HIV-1 infection was observed in cells expressing antisense RNA to the HIV-1 primer-binding site or to the region 5' to the primer-binding site as part of the 3' region of the npt mRNA. Cells expressing the tat mRNA leader sequence in the sense orientation as a precise replacement of the 5' untranslated region of npt mRNA were also resistant to HIV-1. These results indicate that sense and antisense approaches can be used to interfere with HIV-1 multiplication.
1型人类免疫缺陷病毒（HIV-1）主要感染CD4+淋巴细胞和巨噬细胞，并在人类中引发艾滋病。构建了能表达新霉素磷酸转移酶（npt）基因的逆转录病毒载体，使其以反义或正义方向表达HIV-1 RNA的5'序列，并用于转化源自人类CD4+淋巴细胞的MT4细胞系。作为npt mRNA 3'非翻译区的一部分，表达针对HIV-1 tat mRNA前导序列的反义或正义RNA的细胞，对HIV-1感染仍敏感。相比之下，作为npt mRNA 3'区域的一部分，表达针对HIV-1引物结合位点或引物结合位点5'端区域的反义RNA的细胞，对HIV-1感染具有抗性。以精确替换npt mRNA 5'非翻译区的方式，正义方向表达tat mRNA前导序列的细胞，对HIV-1也具有抗性。这些结果表明，正义和反义方法可用于干扰HIV-1的增殖。
