Chlamydial heat shock protein 60 localizes in human atheroma and regulates macrophage tumor necrosis factor-α and matrix metalloproteinase expression
Chlamydial heat shock protein 60 localizes in human atheroma and regulates macrophage tumor necrosis factor-α and matrix metalloproteinase expression
复制标题
DOI：
10.1161/01.cir.98.4.300
复制
发表时间：
1998-07-28
期刊：
影响因子：
37.8
通讯作者：
Libby, P
中科院分区：
文献类型：
--
作者：
Kol, A;Sukhova, GK;Libby, P
背景-最近的证据表明肺炎衣原体与动脉粥样硬化的加重有关。然而，这种药物影响动脉粥样硬化形成的机制仍然知之甚少。衣原体在慢性、持续性感染时产生大量热休克蛋白60（HSP 60），肺炎衣原体主要定位于斑块巨噬细胞内。我们在此验证了动脉粥样硬化包含衣原体HSP 60的假设，这种细菌产物可能刺激巨噬细胞功能，被认为与动脉粥样硬化及其并发症有关，例如产生促炎细胞因子如组织坏死因子-Cu方法和结果：人颈动脉粥样硬化动脉的手术标本（n=19）和正常动脉壁标本（n=7，2条颈动脉和5条动脉）进行了衣原体HSP 60和人HSP 60的存在的免疫化学测试。这些抗原的巨噬细胞定位通过双重免疫染色进行评估。将收获后在无血清条件下维持48小时的小鼠腹腔巨噬细胞与肺炎衣原体HSP 60、人HSP 60或大肠杆菌脂多糖（LPS）孵育。通过ELISA分析在24小时收集用于浓度依赖性实验的培养物上清液和在长达72小时收集用于时间依赖性实验的培养物上清液的TNF-α，并通过明胶酶谱法分析MMP。47%（9/19）的动脉粥样硬化病变显示免疫反应性衣原体HSP 60，89%（17/19）的病例显示人HSP 60。77%（7/9）的病例中衣原体HSP 60与人HSP 60共定位于斑块巨噬细胞内。非动脉粥样硬化样本不含HSP。肺炎衣原体和重组衣原体HSP 60均以浓度和时间依赖性方式诱导小鼠巨噬细胞产生TNF-α。大肠杆菌LPS和人HSP 60产生类似的作用。同样，肺炎衣原体和热休克蛋白诱导MMP的浓度和时间依赖性的方式。热处理废除了肺炎衣原体和热休克蛋白对TNF-α和MMP生产的影响，但它没有改变大肠杆菌LPS诱导这些functions. Conclusions衣原体HSP 60的能力经常与人类HSP 60在人类动脉粥样硬化病变斑块巨噬细胞共定位。衣原体和人HSP 60诱导巨噬细胞产生TNF-α和MMP。衣原体HSP 60可能介导了肺炎衣原体对上述作用的诱导。这种巨噬细胞功能的诱导提供了衣原体感染可能促进动脉粥样硬化形成和加速急性缺血事件的潜在机制。
Background-Recent evidence has implicated Chlamydia pneumoniae in the aggravation of atherosclerosis. However, the mechanisms by which this agent affects atherogenesis remain poorly understood. Chlamydiae produce large amounts of heat shock protein 60 (HSP 60) during chronic, persistent infections, and C pneumoniae localizes predominantly within plaque macrophages, Several studies have furnished evidence that endogenous (human) HSP 60 may play a role in atherogenesis. We tested here the hypothesis that atheroma contains chlamydial HSP 60 and that this bacterial product might stimulate macrophage functions considered relevant to atherosclerosis and its complications, such as production of proinflammatory cytokines as tissue necrosis factor-cu (TNF-alpha) and matrix-degrading metalloproteinases (MMPs).Methods and Results-Surgical specimens of human carotid atherosclerotic arteries (n=19) and normal arterial wall samples (n=7, 2 carotid arteries and 5 aortas) were tested immunohistochemically for the presence of chlamydial HSP 60 and human HSP 60. Macrophage localization of these antigens was assessed by double immunostaining. Murine peritoneal macrophages, maintained in serum-free conditions for 48 hours after harvesting, were incubated with C pneumoniae, chlamydial HSP 60, human HSP 60, or Escherichia coli lipopolysaccharide (LPS). Culture supernatants, collected at 24 hours for concentration-dependence experiments and at up to 72 hours for time-dependence experiments, were analyzed for TNF-alpha by ELISA and for MMP by gelatin zymography. Atherosclerotic lesions showed immunoreactive chlamydial HSP 60 in 47% (9 of 19) of the cases and human HSP 60 in 89% (17 of 19) of the cases. Chlamydial HSP 60 colocalized with human HSP 60 within plaque macrophages in 77% (7 of 9) of the cases. Nonatherosclerotic samples contained neither HSP. Both C pneumoniae and recombinant chlamydial HSP 60 induced TNF-alpha production by mouse macrophages in a concentration- and time-dependent fashion. E coli LPS and human HSP 60 produced similar effects. Similarly, C pneumoniae and HSPs induced MMPs in a concentration- and time-dependent manner. Heat treatment abolished the effect of C pneumoniae and HSPs on both TNF-alpha and MMP production, but it did not alter the ability of E coli LPS to induce these functions.Conclusions-Chlamydial HSP 60 frequently colocalizes with human HSP 60 in plaque macrophages in human atherosclerotic lesions. Chlamydial and human HSP 60 induce TNF-alpha and MMP production by macrophages. Chlamydial HSP 60 might mediate the induction of these effects by C pneumoniae. Induction of such macrophage functions provides potential mechanisms by which chlamydial infections may promote atherogenesis and precipitate acute ischemic events.