Taniguchi M, Nagaoka K, Kunikata T, Kayano T, Yamauchi H, Nakamura S, Ikeda M, Orita K, Kurimoto M
Fujisaki Institute, Hayashibara Biochemical Laboratories Inc., Okayama, Japan.
J Immunol Methods. 1997 Aug 7;206(1-2):107-13. doi: 10.1016/s0022-1759(97)00094-x.
Interleukin-18 (IL-18)/interferon-gamma-inducing factor (IGIF) is a novel cytokine, which is a potent inducer of IFN-gamma production and plays an important role in Th1 responses. In order to develop a specific ELISA for the measurement of human IL-18, we established 13 anti-human IL-18 monoclonal antibodies and characterized them. 7 murine anti-human IL-18 mAbs and 6 rat anti-human IL-18 mAbs were obtained by fusion of splenocytes from mice or rats immunized with human IL-18, with SP2/0 myeloma cells. These antibodies were classified into 4 groups according to competitive binding ELISAs to the human IL-18 molecule. 1 murine mAb and all 6 rat mAbs neutralized IFN-gamma production induced by IL-18. A specific human IL-18 ELISA was developed using two neutralizing mAbs (#125-2H and #159-12B). This ELISA detects human IL-18 with a minimum detection limit of 10 pg/ml, but does not react with heat-denatured human IL-18. The ELISA does not show any cross-reactivity with other cytokines. Using this assay, human IL-18 was measurable in the plasma of leukemia patients. This ELISA would become a powerful tool for investigating the relationship between IL-18 and various diseases or analyzing the control mechanisms of IL-18 production from IL-18 producing cells.
白细胞介素-18（IL-18）/干扰素-γ诱导因子（IGIF）是一种新型细胞因子，是IFN-γ产生的强效诱导剂，在Th1反应中起重要作用。为了开发一种用于检测人IL-18的特异性酶联免疫吸附测定（ELISA），我们制备并鉴定了13株抗人IL-18单克隆抗体。通过用人IL-18免疫的小鼠或大鼠的脾细胞与SP2/0骨髓瘤细胞融合，获得了7株鼠抗人IL-18单克隆抗体和6株大鼠抗人IL-18单克隆抗体。根据对人IL-18分子的竞争性结合ELISA，这些抗体被分为4组。1株鼠单克隆抗体和所有6株大鼠单克隆抗体均能中和IL-18诱导的IFN-γ产生。使用两种中和单克隆抗体（#125-2H和#159-12B）开发了一种特异性人IL-18 ELISA。该ELISA检测人IL-18的最低检测限为10 pg/ml，但不与热变性的人IL-18反应。该ELISA与其他细胞因子无交叉反应。使用该检测方法，可在白血病患者血浆中检测到人IL-18。这种ELISA将成为研究IL-18与各种疾病之间关系或分析IL-18产生细胞中IL-18产生控制机制的有力工具。
