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抗人白细胞介素-4单克隆抗体的特性:在白细胞介素-4酶联免疫吸附测定中的应用以及对白细胞介素-4在B细胞和T细胞上生物活性的差异抑制

Characterisation of monoclonal antibodies to human IL-4: application in an IL-4 ELISA and differential inhibition of IL-4 bioactivity on B cells and T cells.

作者信息

van der Pouw Kraan T, Rensink I, Aarden L

机构信息

Central Laboratory of the Netherlands Red Cross Blood, Transfusion Service (CLB), Amsterdam.

出版信息

Eur Cytokine Netw. 1993 Sep-Oct;4(5):343-9.

PMID:8117935
Abstract

Five murine monoclonal antibodies, raised against E. coli derived human IL-4, were established. All mAb were also reactive with natural IL-4. Competition ELISA experiments revealed that mAb 1,2 and 4 recognized a related epitope on IL-4. mAb 5 and mAb 6 recognized another epitope. Two non-competing mAb were used to develop a sandwich IL-4 ELISA. mAb 5 was used for coating and biotinylated mAb I was used as the second antibody. Intra- and interassay-coefficients were 3.3 and 10.1% respectively. The ELISA is specific for IL-4, rapid and sensitive (the detection limit is 2 pg/ml). The capacities of the antibodies to inhibit IL-4 activity were tested in B cell and T cell assays. All antibodies inhibited IL-4 dependent IgE production by human B lymphocytes. A similar inhibition of IL-4 driven T cell proliferation by the antibodies was observed, with the exception that mAb 4 did not affect the activity of IL-4 on T cells. These results led to the suggestion that B cells make use of another (additional) IL-4 receptor chain.

摘要

制备了五种针对大肠杆菌衍生的人白细胞介素-4(IL-4)产生的鼠单克隆抗体。所有单克隆抗体也与天然IL-4发生反应。竞争酶联免疫吸附测定(ELISA)实验表明,单克隆抗体1、2和4识别IL-4上的一个相关表位。单克隆抗体5和单克隆抗体6识别另一个表位。使用两种非竞争性单克隆抗体开发了一种夹心IL-4 ELISA。单克隆抗体5用于包被,生物素化的单克隆抗体1用作二抗。批内和批间系数分别为3.3%和10.1%。该ELISA对IL-4具有特异性,快速且灵敏(检测限为2 pg/ml)。在B细胞和T细胞测定中测试了抗体抑制IL-4活性的能力。所有抗体均抑制人B淋巴细胞依赖IL-4的IgE产生。观察到抗体对IL-4驱动的T细胞增殖有类似的抑制作用,但单克隆抗体4不影响IL-4对T细胞的活性。这些结果表明B细胞利用了另一条(额外的)IL-4受体链。

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