Qureshi Amer, Zheng Richard, Parlett Terry, Shi Xiaoju, Balaraman Priyadhashini, Cheloufi Sihem, Murphy Brendan, Guntermann Christine, Eagles Peter
Randall Division of Cell and Molecular Biophysics, King's College London, University of London, Guy's Campus, London Bridge, London SE1 1UL, UK.
Biochem J. 2006 Mar 1;394(Pt 2):511-8. doi: 10.1042/BJ20051268.
The chemokine receptors CXCR4 and CCR5 are required for HIV-1 to enter cells, and the progression of HIV-1 infection to AIDS involves a switch in the co-receptor usage of the virus from CCR5 to CXCR4. These receptors therefore make attractive candidates for therapeutic intervention, and we have investigated the silencing of their genes by using ribozymes and single-stranded antisense RNAs. In the present study, we demonstrate using ribozymes that a depletion of CXCR4 and CCR5 mRNAs can be achieved simultaneously in human PBMCs (peripheral blood mononuclear cells), cells commonly used by the virus for infection and replication. Ribozyme activity leads to an inhibition of the cell-surface expression of both CCR5 and CXCR4, resulting in a significant inhibition of HIV-1 replication when PBMCs are challenged with the virus. In addition, we show that small single-stranded antisense RNAs can also be used to silence CCR5 and CXCR4 genes when delivered to PBMCs. This silencing is caused by selective degradation of receptor mRNAs.
趋化因子受体CXCR4和CCR5是HIV-1进入细胞所必需的，HIV-1感染发展为艾滋病涉及病毒共受体使用从CCR5向CXCR4的转变。因此，这些受体成为有吸引力的治疗干预靶点，我们研究了利用核酶和单链反义RNA对其基因进行沉默。在本研究中，我们利用核酶证明，在人外周血单核细胞（PBMCs）中可同时实现CXCR4和CCR5 mRNA的耗竭，PBMCs是该病毒通常用于感染和复制的细胞。核酶活性导致CCR5和CXCR4的细胞表面表达受到抑制，当PBMCs受到病毒攻击时，HIV-1复制受到显著抑制。此外，我们表明，小单链反义RNA在递送至PBMCs时也可用于沉默CCR5和CXCR4基因。这种沉默是由受体mRNA的选择性降解引起的。
