II Medizinische Klinik und Poliklinik, Infektionsambulanz, Klinikum rechts der Isar, Technische Universität München, Munich, Germany.
HIV Med. 2009 Sep;10(8):512-9. doi: 10.1111/j.1468-1293.2009.00723.x. Epub 2009 Jun 1.
Infection with HIV leads to progressive CD4 T-cell loss, resulting in AIDS. Apoptosis is the main mechanism for the loss of infected and bystander cells, but the complex interacting factors inducing and inhibiting apoptosis are not fully understood. Mitochondrial dysfunction is a pivotal step of the apoptotic cascade and can result in reduced mitochondrial membrane potential.
The mitochondrial membrane potential of peripheral blood mononuclear cells (PBMC) was measured by flow cytometry using the dye JC-1 (Molecular Probes Inc). Apoptotic cells were identified using the Annexin V assay (Becton Dickinson GmbH).
The mitochondrial membrane potential of PBMC was significantly decreased and apoptotic cell rate was increased in HIV-infected therapy-naïve patients compared with HIV-negative controls. There was a highly significant correlation between the mitochondrial membrane potential and the rate of apoptosis. CD4 cell count was correlated negatively to the apoptotic rate and positively to the mitochondrial membrane potential.
The JC-1 assay is a sensitive tool to detect changes of mitochondrial membrane potential associated with apoptosis in HIV-infected therapy-naïve patients. We could show in vivo that a reduction of mitochondrial membrane potential is correlated to apoptosis of PBMC, CD4 cell count and HIV viral load during HIV infection.
感染 HIV 会导致 CD4 T 细胞逐渐丧失，从而引发艾滋病。细胞凋亡是感染细胞和旁观者细胞丧失的主要机制，但诱导和抑制细胞凋亡的复杂相互作用因素尚未完全阐明。线粒体功能障碍是细胞凋亡级联反应的关键步骤，可导致线粒体膜电位降低。
使用染料 JC-1（Molecular Probes Inc）通过流式细胞术测量外周血单个核细胞（PBMC）的线粒体膜电位。使用 Annexin V 测定法（Becton Dickinson GmbH）鉴定凋亡细胞。
与 HIV 阴性对照相比，未经治疗的 HIV 感染患者的 PBMC 线粒体膜电位显著降低，凋亡细胞比率增加。线粒体膜电位与凋亡率之间存在高度显著的相关性。CD4 细胞计数与凋亡率呈负相关，与线粒体膜电位呈正相关。
JC-1 测定法是一种灵敏的工具，可用于检测与未经治疗的 HIV 感染患者中凋亡相关的线粒体膜电位变化。我们能够在体内证明，在 HIV 感染过程中，线粒体膜电位的降低与 PBMC、CD4 细胞计数和 HIV 病毒载量的凋亡相关。
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