Kameoka M, Kimura T, Zhong Q, Zheng Y H, Luftig R B, Ikuta K
Section of Serology, Hokkaido University, Sapporo, Japan.
Int Immunol. 1996 Nov;8(11):1687-97. doi: 10.1093/intimm/8.11.1687.
Several recent reports support the hypothesis that apoptosis occurring in leukocytes of human immunodeficiency virus type 1 (HIV-1)-infected individuals is important in progression to AIDS. Specifically, apoptosis of uninfected bystander cells appears critical in the pathogenesis of disease. Here, we present evidence that protease-defective, gp120-containing HIV-1 (L-2) particle preparations specifically induce apoptosis in cells obtained from a subset of promonocytic U937-derived subclones. The rate of apoptosis induction was inversely correlated with the susceptibility of the U937 subclones to wild-type HIV-1 infection. Three types of apoptosis experiments were performed: DNA content analysis by flow cytometry, apoptotic nuclear degradation by fluorescent microscopy and DNA fragmentation analysis by agarose gel electrophoresis. Kinetic analysis revealed that there was a slower induction of apoptosis by L-2 particle preparations than with tumor necrosis factor (TNF)-alpha or anti-Fas antibody. However, there were no significant differences in the initial binding rates of L-2 particles as well as the binding of TNF-alpha or anti-Fas antibody to the U937 subclones. The basal level of protein kinase C activity was higher in high-type subclones compared with low-type subclones. These results suggest that U937 cells can be divided into at least two subpopulations, one that permits a productive HIV-1 infection but is not subjected to L-2 particle preparation-induced apoptosis, while the other poorly replicates HIV-1 and is subjected to L-2 mediated apoptosis, although at a slower rate than found with TNF-alpha or anti-Fas antibody.
最近的几份报告支持这样一种假说,即人类免疫缺陷病毒1型(HIV-1)感染个体白细胞中发生的凋亡在艾滋病进展过程中很重要。具体而言,未感染的旁观者细胞凋亡在疾病发病机制中似乎至关重要。在此,我们提供证据表明,蛋白酶缺陷型、含gp120的HIV-1(L-2)颗粒制剂可特异性诱导从单核细胞U937衍生的亚克隆子集获得的细胞发生凋亡。凋亡诱导率与U937亚克隆对野生型HIV-1感染的易感性呈负相关。进行了三种类型的凋亡实验:通过流式细胞术分析DNA含量、通过荧光显微镜观察凋亡细胞核降解以及通过琼脂糖凝胶电泳分析DNA片段化。动力学分析表明,L-2颗粒制剂诱导凋亡的速度比肿瘤坏死因子(TNF)-α或抗Fas抗体慢。然而,L-2颗粒以及TNF-α或抗Fas抗体与U937亚克隆的初始结合率没有显著差异。与低型亚克隆相比,高型亚克隆中蛋白激酶C活性的基础水平更高。这些结果表明,U937细胞可至少分为两个亚群,一个允许HIV-1进行有效感染,但不受L-2颗粒制剂诱导的凋亡影响,而另一个HIV-1复制能力差,并受到L-2介导的凋亡影响,尽管其速度比TNF-α或抗Fas抗体慢。