Heredia A, Amoroso A, Davis C, Le N, Reardon E, Dominique J K, Klingebiel E, Gallo R C, Redfield R R
Institute of Human Virology, University of Maryland Biotechnology Institute, Baltimore, MD 21201, USA.
Proc Natl Acad Sci U S A. 2003 Sep 2;100(18):10411-6. doi: 10.1073/pnas.1834278100. Epub 2003 Aug 12.
Propagation of R5 strains of HIV-1 on CD4 lymphocytes and macrophages requires expression of the CCR5 coreceptor on the cell surface. Individuals lacking CCR5 (CCR5 Delta 32 homozygous genotype) are phenotypically normal and resistant to infection with HIV-1. CCR5 expression on lymphocytes depends on signaling through the IL-2 receptor. By FACS analysis we demonstrate that rapamycin (RAPA), a drug that disrupts IL-2 receptor signaling, reduces CCR5 surface expression on T cells at concentrations as low as 1 nM. In addition, lower concentrations of RAPA (0.01 nM) were sufficient to reduce CCR5 surface expression on maturing monocytes. PCR analysis on peripheral blood mononuclear cells (PBMCs) showed that RAPA interfered with CCR5 expression at the transcriptional level. Reduced expression of CCR5 on PBMCs cultured in the presence of RAPA was associated with increased extracellular levels of macrophage inflammatory protein (MIP)-1 alpha and MIP-1 beta. In infectivity assays, RAPA suppressed the replication of R5 strains of HIV-1 both in PBMC and macrophage cultures. In total PBMC cultures, RAPA-mediated inhibition of CCR5-using strains of HIV-1 occurred at 0.01 nM, a concentration of drug that is approximately 103 times lower than therapeutic through levels of drug in renal transplant recipients. In addition, RAPA enhanced the antiviral activity of the CCR5 antagonist TAK-779. These results suggest that low concentrations of RAPA may have a role in both the treatment and prevention of HIV-1 infection.
HIV-1 的 R5 毒株在 CD4 淋巴细胞和巨噬细胞上的传播需要细胞表面表达 CCR5 共受体。缺乏 CCR5（CCR5 Delta 32 纯合基因型）的个体表型正常且对 HIV-1 感染具有抗性。淋巴细胞上的 CCR5 表达取决于通过白细胞介素-2 受体的信号传导。通过流式细胞术分析，我们证明雷帕霉素（RAPA），一种破坏白细胞介素-2 受体信号传导的药物，在低至 1 nM 的浓度下即可降低 T 细胞上 CCR5 的表面表达。此外，较低浓度的 RAPA（0.01 nM）足以降低成熟单核细胞上 CCR5 的表面表达。对外周血单核细胞（PBMC）的 PCR 分析表明，RAPA 在转录水平上干扰了 CCR5 的表达。在存在 RAPA 的情况下培养的 PBMC 上 CCR5 表达的降低与巨噬细胞炎性蛋白（MIP）-1α和 MIP-1β细胞外水平的增加有关。在感染性试验中，RAPA 在 PBMC 和巨噬细胞培养物中均抑制了 HIV-1 的 R5 毒株的复制。在总的 PBMC 培养物中，RAPA 介导的对使用 CCR5 的 HIV-1 毒株的抑制作用在 0.01 nM 时出现，该药物浓度比肾移植受者的治疗药物水平低约 103 倍。此外，RAPA 增强了 CCR5 拮抗剂 TAK-779 的抗病毒活性。这些结果表明，低浓度的 RAPA 可能在 HIV-1 感染的治疗和预防中均发挥作用。
