Protein polymorphism of human IL-18 identified by monoclonal antibodies.
Protein polymorphism of human IL-18 identified by monoclonal antibodies.
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单克隆抗体鉴定人IL-18蛋白多态性。
DOI：
10.3892/ijmm.8.5.585
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发表时间：
2001
影响因子：
5.4
通讯作者：
K. Toyoshima
中科院分区：
文献类型：
--
作者：
T. Seya;M. Matsumoto;I. Shiratori;Y. Fukumori;K. Toyoshima
制备了6株抗人“功能失活”重组IL-18的单抗，用其中两株单抗(#21和#132)建立了检测IL-18“功能失活”形式的酶联免疫吸附试验，并用人血浆和巨噬细胞(MP)检测了IL-18蛋白的失活形式。在GM-CSF处理6天的单核细胞，即MP中，mAb#21通过免疫印迹识别IL-18前体(24 KDa)和48 kDa二聚体。相比之下，只有24 kDa的物种被检测到是一条相对较弱的带，带有商业上的抗“活性”IL-18的mAb。未成熟单核细胞中未检测到IL-18类物质。因此，在MP中产生了二聚体IL-18，并可被我们建立的mAb检测到。在正常人和患者的血浆中，也检测到了#21可识别的IL-18，其水平与商业上可用于测定具有功能活性的IL-18的试剂盒的水平不一致。我们将前者定为2型，后者定为1型。值得注意的是，所有志愿者都检测到了IL-18 1型，而大约30%的健康受试者检测到了2型，而且血浆中2型的水平(10-100 ng/ml)高于1型(0.02-0.55 ng/ml)。在特应性皮炎患者中，1型的平均值(200 ng/ml)高于正常人(0.122 ng/ml)和肺癌患者(0.113 ng/ml)。特应性皮炎中高1型的产生可能与免疫调节状态有关。IL-182型在这些人群中的水平和频率没有显著变化。因此，在单核细胞-MP分化过程中会产生大量的2型物种，它们在血浆中的水平和频率没有变化，与1型的水平无关。
Six mAbs were raised against human "functionally inactive" recombinant IL-18, ELISA for determination of "functionally inactive" forms of IL-18 were established using two of these mAbs (#21 and #132), and inactive species of IL-18 protein were examined with human blood plasma and macrophages (Mp). In 6-day GM-CSF-treated monocytes, namely Mp, the mAb #21 recognized the IL-18 proform (24 kDa) and a 48 kDa dimer by immunoblotting. In contrast, only the 24 kDa species was detected as a relatively faint band with a commercial mAb against "active" IL-18. No IL-18 species was detected in premature monocytes. Thus, the dimeric IL-18 was produced in Mp and detectable with the mAb we established. In blood plasma of normal subjects and patients, the #21-recognizable IL-18 was also detected by ELISA, the levels of which were not consistent with those obtained with the commercially available kit for determination of "functionally active" IL-18. We designated the former as type 2 and the latter as type 1. Strikingly, IL-18 type 1 was detected in all volunteers while type 2 was detected in approximately 30% of healthy subjects, and the levels of type 2 were high (10-100 ng/ml) compared to those of type 1 (0.02-0.55 ng/ml) in their blood plasma. In patients with atopic dermatitis, the mean value of type 1 was high (200 ng/ml) compared to those of normal subjects (0.122 ng/ml) and patients with lung cancer (0.113 ng/ml). Production of high type 1 may be associated with an immunomodulatory state in atopic dermatitis. The levels and frequencies of IL-18 type 2 were not significantly changed among these populations. Hence, large amounts of type 2 species are produced in monocyte-Mp differentiation, and their levels and frequencies are unchanged in blood plasma irrespective of the levels of type 1.