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

通过原病毒 DNA 水平和 T 细胞更新评估残留病毒复制，以优化 HAART。

基本信息

批准号：
14570422
负责人：
YOSHIMURA Kazuhisa
金额：
$ 2.18万
依托单位：
Kumamoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570422/
关键词：
HIV-1 residual replication Turnover HAART proviral DNA CD4^+ T cell

项目摘要

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.

使用高度灵敏的检测方法来检测前病毒 DNA (pDNA) 和 T 淋巴细胞的更新，我们正在尝试优化 HAART，以最大程度地减少血浆病毒血症患者体内的残留病毒。使用新型超敏巢式 PCR 测量了 HIV-1 阳性患者的 PBMC 中 LTR 区域的 pDNA 水平。常规第一次 PCR 后进行定量实时 PCR 荧光测定以检测 pDNA。我们还通过四色流式细胞术分析测量核抗原 Ki-67 来研究 CD4^+ 和 CD8^+ T 细胞更新。我们测量了来自病毒血症或无病毒血症患者的 340 个样本的 PBMC 中的 HIV pDNA 水平。在HAART检测不到血浆病毒血症的患者中，pDNA检测不到的患者的CD4^+ T细胞计数和CD4/8比值显着高于pDNA可检测到的患者（p<0.01）。我们还跟踪了一名患者，该患者可以根据 pDNA 和 T 细胞更新的结果选择治疗优化。当 HAART 方案优化为更有效的组合时，观察到 pDNA 水平显着下降。添加依非韦伦 (EFV) 30 个月后，观察到 CD4^+ 子集加速更新的正常化和 pDNA 的下降。我们的研究表明，pDNA 和 T 细胞更新的测量适用于评估患者体内 HIV-1 的残留复制。通过提供这些结果并明智地选择有效的抗逆转录病毒药物组合，可以实现长期成功的治疗。