细胞因子释放综合征的2种体外检测方法比较研究  

Comparative study of two in vitro experimental methods detecting cytokine release syndrome

作　　者：徐静怡 宇家仪 马璟 XU Jing-yi;YU Jia-yi;MA Jing(China State Institute of Pharmaceutical Industry,Shanghai 201203,China;Shanghai InnoStar Biotech Co.,Ltd.,Shanghai 201203,China)

机构地区：[1]中国医药工业研究总院,上海201203 [2]上海益诺思生物技术股份有限公司,上海201203

出　　处：《中国新药杂志》2021年第24期2256-2261,共6页Chinese Journal of New Drugs

基　　金：国家“重大新药创制”科技重大专项资助项目(2019ZX09732001-020)。

摘　　要：目的:对固相法及液相法2种体外细胞因子释放实验方法进行比较研究,为评价新药是否引起细胞因子释放方法学选择提供依据。方法:用2种不同方法与外周血单核细胞共同孵育24 h,流式细胞磁珠微阵列法检测白介素-1β(IL-1β)、IL-6、IL-8、肿瘤坏死因子-α(TNF-α)这4种细胞因子的表达水平。结果:固相法中,IL-1β仅在抗CD3单克隆抗体(OKT3)的5μg·孔^(-1)有显著性释放(P<0.05);IL-6在2种抗体中均有显著性释放(P<0.05);OKT3所有浓度及CD28超级激动剂TGN1412的5μg·孔^(-1) IL-8与TNF-α有显著性释放(P<0.05)。液相法中,OKT3的1和5μg·孔^(-1)、TGN1412的5μg·孔^(-1) IL-6有显著性释放(P<0.05);TGN1412的5μg·孔^(-1) IL-8有显著性释放(P<0.05);OKT3所有浓度及TGN1412的5μg·孔^(-1) TNF-α有显著性释放(P<0.05)。结论:固相法更适用于T细胞受体的单克隆抗体药物,在临床前评价中其结果可信度更高;液相法操作简单,并且其模拟的环境更接近于人体,对于靶点不在T细胞上的单克隆抗体有更高的灵敏度。Objective:A comparative study of two in vitro experimental methods detecting cytokine release was conducted for evaluating methodological choices on whether new drugs cause cytokine release.Methods:Peripheral blood mononuclear cells were incubated with two different methods for 24 hours,after which the expression levels of four cytokines:IL-1β,IL-6,IL-8 and TNF-αwere detected by cytometric bead assay.Results:In solid phase method,IL-1βwas significantly released only in OKT3 at 5μg·well^(-1)(P<0.05).Significant release of IL-6 was found in two antibodies(P<0.05).There was a significant release of IL-8 and TNF-αat all concentrations of OKT3 and 5μg·well^(-1) of TGN1412(P<0.05).While in liquid phase method,significant release of IL-6 happened in 1 and 5μg·well^(-1) of OKT3,as well as 5μg·well^(-1) of TGN1412(P<0.05).There was a significant release of IL-8 at 5μg·well^(-1) of TGN1412(P<0.05).Significant release of TNF-αwas found in all concentrations of OKT3 and 5μg·well^(-1) of TGN1412(P<0.05).Conclusion:The solid phase method is more suitable for T cell receptor monoclonal antibody drugs,the results of which are more reliable in preclinical evaluation.The liquid phase method is simpler to operate,and its simulated environment is closer to human body,but it has higher sensitivity for monoclonal antibodies whose target is not on T cells.

关 键 词：细胞因子释放综合征 CD3单克隆抗体 CD28超级激动剂 外周血单核细胞 流式细胞磁珠微阵列法 

分 类 号：R917[医药卫生—药物分析学]