Tumor necrosis factor-α in whole blood cultures of preeclamptic patients and healthy pregnant and nonpregnant women
Tumor necrosis factor-α in whole blood cultures of preeclamptic patients and healthy pregnant and nonpregnant women
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DOI：
10.1081/prg-200030334
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发表时间：
2004-01-01
影响因子：
1.5
通讯作者：
Wallenburg, HCS
中科院分区：
文献类型：
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作者：
Beckmann, I;Ben Efraim, S;Wallenburg, HCS
目的：肿瘤坏死因子-α（TNF-alpha）被认为是过度内皮激活和损伤的可能介质，而过度内皮激活和损伤是先兆子痫的关键发病机制。我们使用了12例重度先兆子痫患者和12例健康孕妇和非孕妇的全血细胞培养物，以确定未受刺激的白细胞释放TNF-α作为其激活状态的衡量标准，以及其对脂多糖（LPS）刺激的反应作为其启动状态的指标。研究方法：在无LPS和有LPS的情况下培养血液，并在培养6小时和24小时后通过酶联免疫测定法测量TNF-α的释放。进行白细胞分类计数，并计算每105个单核细胞的TNF-α值。结果如下：在未经刺激的全血培养，TNF-α释放后6小时的培养是相似的，在所有三组，但24小时后，TNF-α浓度的培养上清液中先兆子痫患者显着高于从血压正常的孕妇血液中获得的值。在LPS刺激的血培养中，TNF-α的释放在6小时的培养时间达到最大值，先兆子痫妇女的TNF-α浓度显著低于两个对照组。我们在另一项实验中表明，与低剂量LPS预激活后TNF-α的释放相比，高剂量LPS预激活后的强烈LPS激发导致TNF-α的释放减少。结论：观察到先兆子痫中白细胞自发释放TNF-α的高能力表明疾病过程激活了产生TNF-α的白细胞。如在先兆子痫患者的血培养中观察到的，由于TNF-α泄漏导致的白细胞的预活化和耗竭可导致对TNF-α诱导剂LPS的反应降低。
Objectives: Tumor necrosis factor-alpha (TNF-alpha) is recognized as a likely mediator of the excessive endothelial activation and injury that is a key pathogenetic mechanism of preeclampsia. We used whole blood cell cultures from 12 patients with severe preeclampsia and from 12 healthy pregnant and nonpregnant women to determine the release of TNF-alpha by unstimulated leukocytes as a measure of their state of activation, and their response to stimulation with lipopolysaccharide (LPS) as an indicator of their state of priming. Methods: Blood was cultivated without and with LPS, and TNF-alpha release was measured after six and 24 hours of cultivation by enzyme-linked immunoassays. Differential leukocyte counts were performed, and TNF-alpha values calculated per 105 monocytes. Results: In unstimulated whole blood cultures, TNF-alpha release after six hours of cultivation was similar in all three groups; but after 24 hours, TNF-alpha concentrations in culture supernatants from preeclamptic patients were significantly higher than were values obtained in blood from normotensive pregnant women. In LPS-stimulated blood cultures with a maximum of TNF-alpha release at six hours cultivation time, TNF-alpha concentrations were significantly lower in preeclamptic women than they were in both control groups. We showed in an additional experiment that a strong LPS challenge following preactivation with high doses of LPS resulted in reduced release of TNF-alpha compared with release of TNF-alpha following preactivation with low doses of LPS. Conclusions: The observed high capacity for spontaneous TNF-alpha release by leukocytes in preeclampsia indicates activation of TNF-alpha producing leukocytes by the disease process. Preactivation and exhaustion of leukocytes by leakage of TNF-alpha could lead to the reduced response to TNF-alpha inducer LPS as observed in blood cultures from preeclamptic patients.