Gerdes J, Herzbeck H, Schlüter C, Flad H D
Department of Immunology and Cell Biology, Forschungsinstitut Borstel, FRG.
Lymphokine Res. 1989 Fall;8(3):239-43.
An immunoenzymatic detection method of in-situ hybridization reactions for interleukin 1 (IL-1) beta was established, using sulphonated probes. As model system we used unstimulated and lipopolysaccharide (LPS) stimulated peripheral blood mononuclear cells (PBMNC). After hybridization, sulphone groups were targeted with a monoclonal antibody, and bound antibody was visualized by the alkaline phosphatase anti-alkaline phosphatase (APAAP) method. In unstimulated PBMNC both the control and the IL-1 beta specific c-DNA probes were negative, whilst a large proportion of LPS treated PBMNC was positive with the sulphonated IL-1 beta plasmid only. This method may be a powerful alternative to radio-isotopic labeling. Since the entire procedure can be performed within one day, it may be applied for routine diagnostic purposes.
建立了一种使用磺化探针的白细胞介素1（IL-1）β原位杂交反应的免疫酶检测方法。作为模型系统，我们使用了未刺激的和脂多糖（LPS）刺激的外周血单个核细胞（PBMNC）。杂交后，用单克隆抗体靶向磺酸基团，并通过碱性磷酸酶抗碱性磷酸酶（APAAP）方法使结合的抗体可视化。在未刺激的PBMNC中，对照和IL-1β特异性c-DNA探针均为阴性，而仅磺化的IL-1β质粒处理的LPS的大部分PBMNC为阳性。该方法可能是放射性同位素标记的有力替代方法。由于整个过程可以在一天内完成，因此可用于常规诊断目的。
