Human macrophages produce dimeric forms of IL-18 which can be detected with monoclonal antibodies specific for inactive IL-18.
Human macrophages produce dimeric forms of IL-18 which can be detected with monoclonal antibodies specific for inactive IL-18.
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人类巨噬细胞产生二聚体形式的 IL-18，可以使用非活性 IL-18 特异性单克隆抗体进行检测。
DOI：
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发表时间：
2001
期刊：
影响因子：
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通讯作者：
T. Seya
中科院分区：
文献类型：
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作者：
S. Kikkawa;M. Matsumoto;K. Shida;Y. Fukumori;K. Toyoshima;T. Seya
我们建立了两株能识别人功能失活的重组IL-18的单抗，并用人单核巨噬细胞(M Phi)检测了IL-18蛋白的多态性。在GM-CSF处理的第6天，IL-18识别的mAb21检测到24 kDa的IL-18前体和48 kDa的蛋白，随着成熟期的增加而逐渐增加。大多数24 kDa和48 kDa的形式几乎检测不到，其他mAbs识别“活性的”IL-18。包括Toll刺激剂在内的任何试剂都不能上调M Phi中的这些IL-18群体。用阴离子交换色谱柱分离出21种可识别的IL-18分子，并用人淋巴细胞和IL-12检测它们的干扰素-γ诱导活性。这些IL-18物种几乎没有检测到已知的活性。经M Phi蛋白水解酶处理后，产生了一种18 kDa的形式来表达干扰素-γ诱导活性，尽管其活性远弱于对照的活性IL-18。这些观察结果表明，随着单核细胞-Mphi的分化，产生了大量不同的IL-18物种，其中大多数IL-18物种即使在蛋白水解性18-kDa转化后，就已报道的IL-18功能而言也是“不活跃的”。
We established two monoclonal antibodies (mAbs) which specifically recognize human 'functionally inactive' recombinant IL-18, and IL-18 protein polymorphism was examined using human monocytes and macrophages (M phi). In 6 day GM-CSF-treated M phi, an 'inactive' IL-18-recognizing mAb 21 detected the IL-18 proform (24 kDa) and a 48-kDa protein, which were gradually increased concomitant with maturation stage. Majority of the 24- and 48-kDa forms were barely detectable with other mAbs recognizing 'active' IL-18. No reagents including Toll stimulators up-regulated these IL-18 populations in M phi. The 21-recognizable IL-18 species were separated using an anion-exchanger column and their IFN gamma-inducing activity was assessed with human lymphocytes plus IL-12. Virtually no as yet known activity was detected with these IL-18 species. After processed with M phi proteases, an 18-kDa form was generated to express the IFN gamma-inducing activity, although the activity was far weaker than that of control 'active' IL-18. These observations suggested that large amounts of various IL-18 species are produced with monocyte-M phi differentiation and most of these IL-18 species are functionally 'inactive' in terms of the reported IL-18 function even after proteolytic 18-kDa conversion.
DOI：
10.1073/pnas.96.5.2256
发表时间：
1999-03-02
影响因子：
11.1
作者：
Puren, AJ;Fantuzzi, G;Dinarello, CA
通讯作者：
Dinarello, CA