检测单核细胞体外肿瘤坏死因子α产生的新方法。
Novel method for detection of ex vivo tumor necrosis factor alpha production by monocytes.
作者信息
Suzuki Kikumi, Koyama Takatoshi, Kobayashi Shizuko, Kobayashi Koji, Inagaki Koji, Abe Yoshiko, Kuriyama Kiyoshi, Shiba Kiyoko
机构信息
Department of Molecular Physiology, Kyoritsu College of Pharmacy Tokyo, Japan.
出版信息
J Clin Lab Anal. 2002;16(6):273-8. doi: 10.1002/jcla.10050.
Tumor necrosis factor alpha (TNF-alpha) has come into recent focus as a proinflammatory cytokine derived from monocytes/macrophages. We developed a novel system (the SEK-5001 system) for measurement of ex vivo production of TNF-alpha stimulated by lipopolysaccharide (LPS) as a marker of immune function. In the present study, we evaluated the performance of this system as a diagnostic tool. Furthermore, we compared TNF-alpha levels with data from other immune function tests, including lymphocyte blast formation test and differential leukocyte counts. Incubation of whole blood with a stimulation of low-dose LPS (100 EU/mL blood) for 4 hr at 37 degrees C gave acceptable results. After incubation, plasma TNF-alpha levels were determined by enzyme-linked immunosorbent assay (ELISA). The specificity, reproducibility, and recovery of the SEK-5001 system were excellent. No correlation between TNF-alpha levels and total leukocyte counts was found. Lymphocyte blast formation test and monocyte counts, however, were correlated with TNF-alpha levels in blood from patients with hematological malignancy and aplastic anemia before treatment. This assay system may potentially be clinically applicable to assess in vivo immune function.
肿瘤坏死因子α（TNF-α）作为一种源自单核细胞/巨噬细胞的促炎细胞因子，近来受到关注。我们开发了一种新型系统（SEK-5001系统），用于测量脂多糖（LPS）刺激下TNF-α的体外产生量，以此作为免疫功能的标志物。在本研究中，我们评估了该系统作为诊断工具的性能。此外，我们将TNF-α水平与其他免疫功能测试的数据进行了比较，包括淋巴细胞转化试验和白细胞分类计数。将全血与低剂量LPS（100 EU/mL血液）在37℃下刺激孵育4小时，得到了可接受的结果。孵育后，通过酶联免疫吸附测定（ELISA）测定血浆TNF-α水平。SEK-5001系统的特异性、可重复性和回收率都非常出色。未发现TNF-α水平与白细胞总数之间存在相关性。然而，淋巴细胞转化试验和单核细胞计数与血液系统恶性肿瘤和再生障碍性贫血患者治疗前血液中的TNF-α水平相关。该检测系统可能在临床上适用于评估体内免疫功能。
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