A Cell Assay for Detecting Anti-PEG Immune Response against PEG-Modified Therapeutics
A Cell Assay for Detecting Anti-PEG Immune Response against PEG-Modified Therapeutics
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DOI：
10.1007/s11095-018-2505-3
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发表时间：
2018-11-01
影响因子：
3.7
通讯作者：
Ishida, Tatsuhiro
中科院分区：
文献类型：
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作者：
Shimizu, Taro;Abu Lila, Amr S.;Ishida, Tatsuhiro
聚乙二醇化蛋白质和纳米药物的免疫原性是其在临床环境中开发和使用的潜在障碍。本研究的目的是建立一种利用流式细胞术检测PEG化蛋白和/或纳米药物抗PEG免疫的方法。方法荧光标记的mPEG修饰的脂质体与HIK-G11细胞、PEG特异性杂交瘤细胞、用流式细胞术检测PEG化治疗剂的免疫原性。修饰的脂质体与HIK-G11细胞有效结合。在未标记的mPEG修饰的脂质体或mPEG修饰的卵清蛋白（OVA）的存在下，荧光标记的mPEG修饰的脂质体的这种染色被显著抑制，但不聚甘油修饰的脂质体。此外，我们发现mPEG修饰的脂质体，高免疫原性，引起PEG特异性细胞的增殖，而羟基PEG修饰的脂质体，免疫原性较低，几乎没有引起。此外，静脉注射mPEG修饰的脂质体后，在脾细胞中的PEG特异性细胞的百分比，通过流式细胞术测定，以及对应的抗PEG抗体的生产水平，通过ELISA. ConclusionPEG特异性B细胞分析，我们介绍可能成为一个有用的方法来检测抗PEG免疫反应对聚乙二醇化的治疗剂，并澄清抗PEG免疫反应的机制。
Purpose Immunogenicity of PEGylated proteins and nanomedicines represents a potential impediment against their development and use in clinical settings. The purpose of this study is to develop a method for detecting anti-PEG immunity of PEGylated proteins and/or nanomedicines using flow cytometry.Methods The binding of fluorescence-labeled mPEG-modified liposomes to HIK-G11 cells, PEG-specific hybridoma cells, or spleen cells was evaluated by flow cytometry for detecting immunogenicity of PEGylated therapeutics.Results The fluorescence-labeled methoxy PEG (mPEG)-modified liposomes were efficiently bound to HIK-G11 cells. Such staining with fluorescence-labeled mPEG-modified liposomes was significantly inhibited in the presence of either non-labeled mPEG-modified liposomes or mPEG-modified ovalbumin (OVA) but not polyglycerol-modified liposomes. In addition, we found that mPEG-modified liposomes, highly immunogenic, caused proliferation of PEG-specific cells, while hydroxyl PEG-modified liposomes, less immunogenic, scarcely caused. Furthermore, after intravenous injection of mPEG-modified liposomes, the percentage of PEG-specific cells in the splenocytes, as determined by flow cytometry, corresponded well with the production level of anti-PEG antibodies, as determined by ELISA.Conclusions PEG-specific B cell assay we introduced may become a useful method to detect an anti-PEG immune response against PEGylated therapeutics and clarify the mechanism for anti-PEG immune responses.