Heredia Alonso, Davis Charles, Amin Mohammed N, Le Nhut M, Wainberg Mark A, Oliveira Maureen, Deeks Steven G, Wang Lai-Xi, Redfield Robert R
aInstitute of Human Virology, University of Maryland School of Medicine, Baltimore, Maryland, USA bMcGill University AIDS Centre, Lady Davis Institute, Jewish General Hospital, Montreal, Quebec, Canada cSan Francisco VA Medical Center and University of California, San Francisco, California, USA.
AIDS. 2014 Jan 28;28(3):317-23. doi: 10.1097/QAD.0000000000000168.
The M184V mutation in the HIV-1 reverse transcriptase gene is frequent (>50%) in patients, both in resource-rich and resource-limited countries, conferring high-level resistance (>100-fold) to the cytosine analog reverse transcriptase inhibitors lamivudine and emtricitabine. The reverse transcriptase enzyme of M184V HIV-1 mutants has reduced processivity, resulting in reduced viral replication, particularly at low deoxynucleotide (dNTP) levels. We hypothesized that lowering intracellular dNTPs with resveratrol, a dietary supplement, could interfere with replication of M184V HIV-1 mutants.
Evaluation of the activity of resveratrol on infection of primary peripheral blood lymphocytes by wild-type and M184V mutant HIV-1. We assayed both molecular clones and primary isolates of HIV-1, containing M184V alone and in combination with other reverse transcriptase mutations. Viral infection was quantified by p24 ELISA and by quantitative real-time PCR analysis. Cell viability was measured by colorimetric 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assays.
In virus-infectivity assays, resveratrol did not inhibit replication of wild-type NL4-3 (resveratrol EC50 > 10 μmol/l), but it inhibited NL4-3 184V mutant (resveratrol EC50 = 5.8 μmol/l). These results were confirmed by real-time PCR analysis of early and late products of reverse transcription. Resveratrol inhibited molecular clones and primary isolates carrying M184V, alone or in combination with other reverse transcriptase mutations (resveratrol EC50 values ranging from 2.5 to 7.7 μmol/l).
Resveratrol inhibits HIV-1 strains carrying the M184V mutation in reverse transcriptase. We propose resveratrol as a potential adjuvant in HIV-1 therapy, particularly in resource-limited settings, to help control emtricitabine-resistant M184V HIV-1 mutants.
在资源丰富和资源有限的国家，HIV-1逆转录酶基因中的M184V突变在患者中都很常见（>50%），该突变赋予对胞嘧啶类似物逆转录酶抑制剂拉米夫定和恩曲他滨高水平耐药性（>100倍）。M184V HIV-1突变体的逆转录酶具有降低的持续合成能力，导致病毒复制减少，尤其是在低脱氧核苷酸（dNTP）水平时。我们推测，用白藜芦醇（一种膳食补充剂）降低细胞内dNTP水平可能会干扰M184V HIV-1突变体的复制。
评估白藜芦醇对野生型和M184V突变型HIV-1感染原代外周血淋巴细胞活性的影响。我们检测了HIV-1的分子克隆和原代分离株，这些毒株单独携带M184V或与其他逆转录酶突变共同携带M184V。通过p24 ELISA和定量实时PCR分析对病毒感染进行定量。通过比色法3-[4,5-二甲基噻唑-2-基]-2,5-二苯基溴化四氮唑（MTT）测定法测量细胞活力。
在病毒感染性试验中，白藜芦醇不抑制野生型NL4-3的复制（白藜芦醇EC50>10 μmol/l），但它抑制NL4-3 184V突变体（白藜芦醇EC50 = 5.8 μmol/l）。这些结果通过对逆转录早期和晚期产物的实时PCR分析得到证实。白藜芦醇抑制携带M184V的分子克隆和原代分离株，无论其单独携带还是与其他逆转录酶突变共同携带（白藜芦醇EC50值范围为2.5至7.7 μmol/l）。
白藜芦醇抑制逆转录酶携带M184V突变的HIV-1毒株。我们提出白藜芦醇可作为HIV-1治疗中的一种潜在佐剂，尤其是在资源有限的环境中，以帮助控制对恩曲他滨耐药的M184V HIV-1突变体。
