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Immunological studies on Kawasaki disease: II. Isolation and characterization of an immunosuppressive factor in acute phase sera.

机译:川崎病的免疫学研究:II。急性期血清中免疫抑制因子的分离和鉴定。

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摘要

Acute phase serum from a patient with Kawasaki disease possessed strong inhibitory activity for the proliferative response of Con A-stimulated peripheral blood mononuclear cells. The inhibitor was fractionated step-wise by means of DEAE-Sephadex A-50 ion exchange chromatography followed by Sephadex G-100 gel filtration and high performance liquid chromatography. A major protein of 140 kD with 2000 times greater inhibitory activity than the original serum was identified in the final fraction. Immunization of mice with this fraction resulted in the production of three hybridoma clones secreting monoclonal antibodies (MoAb) of IgG1 class which blocked the inhibitory activity of the fraction. Two of these MoAb recognized the same epitope of the inhibitory factor, while the remaining MoAb was directed to a different epitope. Western blot analysis of acute phase sera by the MoAb demonstrated the presence of 140 kD molecules in 43 of 55 patients.
机译:川崎病患者的急性期血清对Con A刺激的外周血单个核细胞的增殖反应具有很强的抑制活性。通过DEAE-Sephadex A-50离子交换色谱法逐步分离抑制剂,然后进行Sephadex G-100凝胶过滤和高效液相色谱法。在最终馏分中鉴定出140 kD的主要蛋白质,其抑制活性是原始血清的2000倍。用该级分免疫小鼠会产生三个杂交瘤细胞克隆,这些克隆分泌IgG1类单克隆抗体(MoAb),从而阻断了该级分的抑制活性。这些MoAb中的两个识别抑制因子的相同表位,而其余MoAb指向不同的表位。 MoAb对急性期血清的蛋白质印迹分析表明,在55名患者中的43名患者中存在140 kD分子。

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