Evaluation of residual viral replication by proviral DNA level and T cell turnover for optimization of HAART.

使用一种高灵敏度的检测前病毒DNA(PDNA)和T淋巴细胞周转的方法，我们正试图优化HAART，以最大限度地减少无法检测到的血浆病毒血症患者的残留病毒。用一种新型的超敏套式聚合酶链式反应检测HIV-1阳性患者外周血单核细胞中LTR区的PDNA水平。常规第一次聚合酶链式反应后，采用实时荧光定量聚合酶链式反应检测PDNA。用四色流式细胞仪检测细胞核抗原Ki-67，观察CD4~+和CD8~+T细胞的转化率。我们检测了340例病毒症或无病毒症患者外周血单个核细胞中的HIV PDNA水平。在HAART检测不出血浆病毒血症的患者中，PDNA检测不到的患者外周血中CD4~+T细胞计数和CD_4/8比值显著高于可检测到的患者(P&lt；0.01)。我们还跟踪了一位患者，他可以根据PDNA和T细胞周转率的结果选择最佳治疗方案。当HAART方案被优化为更有效的组合时，观察到PDNA水平显著下降。加入EFV治疗30个月后，CD_4~+亚群加速周转恢复正常，PDNA下降。我们的研究表明，PDNA和T细胞周转率的测量适合于评估HIV-1在患者体内的残留复制。通过向这些结果提供有效的抗逆转录病毒药物组合的知情选择，可以实现长期的成功治疗。

Using a highly sensitive assay to detect proviral DNA(pDNA) and the turnover of T lymphocytes, we are attempting to optimize HAART to minimize residual viruses in patients with undetectable plasma viremia. The pDNA levels in PBMCs from HIV-1 positive patients were measured in the LTR region using a novel hypersensitive nested PCR. Quantitative real-time PCR fluorogenic assay was performed to detect pDNA after conventional first PCR. We also investigated CD4^+ and CD8^+ T cell turnover by measuring the nuclear antigen Ki-67 with four-color flow cytometry analysis. We measured the HIV pDNA level in PBMCs of 340 samples from viremic or aviremic patients. Among the patients with undetectable plasma viremia by HAART, the CD4^+ T cell count and CD4/8 ratio were significantly higher in patients with undetectable pDNA than in patients with detectable pDNA(p<0.01). We also followed a patient who has an option to choose treatment optimization based on results of pDNA and T-cell turnover. Significant decline of the pDNA level was observed when the regimen of HAART was optimized to a more potent combination. Both normalization of accelerated turnover of CD4^+ subset and decline of pDNA were observed after 30 months from the addition of efavirenz(EFV). Our study suggests that the measurement of both pDNA and T-cell turnover is suitable for evaluating the residual replication of HIV-1 in patients. Long-term successful treatment is achievable by providing these results with an informed choice of a potent combination of antiretrovirals.