Biotherapeutics Group, National Institute for Biological Standards and Control, Potters Bar, Hertfordshire EN6 3QG, UK.
J Immunol Methods. 2011 Aug 31;371(1-2):134-42. doi: 10.1016/j.jim.2011.06.022. Epub 2011 Jun 30.
Two methods for predicting the risk of pro-inflammatory clinical infusion reactions during monoclonal antibody therapy were evaluated. In the first, the antibody of interest is immobilised by air-drying onto 96-well plates prior to the addition of human peripheral blood mononuclear cells (PBMCs). In the second, the antibody is added in aqueous phase to a co-culture of human PBMCs and human endothelium-derived cells. In both methods the cells are incubated with the antibody to allow the accumulation of pro-inflammatory cytokines, quantified by enzyme-linked immunosorbent assay (ELISA). The antibodies associated with clinical infusion reactions, Herceptin, Campath-1H and TGN1412, gave the largest responses taking into account the data for all readouts (tumour necrosis factor-α, TNF, interleukin-6, IL-6, IL-8, IL-2 and cell proliferation) for both methods. Overall, the antibodies tested could be ranked as follows: Tysabri<Avastin<Herceptin<Campath-1H<TGN1412, with only the "superagonistic" CD28 monoclonal antibody (TGN1412) stimulating IL-2 release and cell proliferation.
评估了两种预测单克隆抗体治疗中促炎临床输注反应风险的方法。第一种方法是将感兴趣的抗体通过空气干燥固定在 96 孔板上，然后加入人外周血单核细胞 (PBMC)。第二种方法是将抗体在水相中添加到人 PBMC 和人内皮细胞衍生细胞的共培养物中。在这两种方法中，细胞与抗体孵育以允许促炎细胞因子积累，通过酶联免疫吸附试验 (ELISA) 定量。考虑到两种方法的所有读数（肿瘤坏死因子-α、TNF、白细胞介素-6、IL-6、白细胞介素-8、IL-2 和细胞增殖），与临床输注反应相关的抗体，赫赛汀、坎帕斯-1H 和 TGN1412 产生了最大的反应。总体而言，测试的抗体可以按以下顺序排列：Tysabri<Avastin<Herceptin<Campath-1H<TGN1412，只有“超激动”CD28 单克隆抗体 (TGN1412) 刺激白细胞介素-2 释放和细胞增殖。
