基于小鼠腘窝淋巴结模型对疫苗用辅料蔗糖中不溶性微粒激发免疫应答风险的评估  

Immune Response Risk Assessment of Insoluble Particles in Sucrose for Vaccine Based on PLNA Mouse Model

作　　者：王珏[1] 江颖 沈雁[3] 肖新月[1] 杨锐[1] 孙会敏[1] WANG Jue;JIANG Ying;SHEN Yan;XIAO Xin-yue;YANG Rui;SUN Hui-min(NMPA Key Laboratory for Quality Research and Eraluation of Pharmaceutical Excipients,National Institutes for Food and Drug Control,Beijing 100050,China;Eivaluation and Verification Center for Medical Equipment and Cosmetics of Shanghai,Shanghai 200020,China;NMPA Key Laboratory for Research and Evaluation of Pharmaceutical Preparations and Excipients,China Pharmaceutical University,Nanjing 210009,China)

机构地区：[1]国家药品监督管理局药用辅料质量研究与评价重点实验室,中国食品药品检定研究院,北京100050 [2]上海市医疗器械化妆品审评核查中心,上海200001 [3]国家药品监督管理局药物制剂及辅料研究与评价重点实验室,中国药科大学,江苏南京210009

出　　处：《药物生物技术》2022年第3期234-238,共5页Pharmaceutical Biotechnology

摘　　要：疫苗安全性近年来受到极大关注,疫苗用辅料的安全性也因此受到挑战。尽管生物制品中不溶性微粒受到严格的监管,但关于其是否来源于辅料或辅料在其中贡献大小的研究仍很少。文章为探究疫苗中常用保护剂蔗糖引入的不溶性微粒及其安全性风险,用超滤法提取制备了蔗糖中的不溶性微粒并用纳米颗粒追踪技术对其进行颗粒计数和粒径分析,用扫描电镜对其进行微观形貌表征,结果表明该微粒粒径为100~300 nm,不同厂家蔗糖的不溶性微粒在大小和数量上均存在差异。采用小鼠腘窝淋巴结模型对蔗糖中不溶性微粒激发免疫应答风险进行评估,小鼠足趾厚度、淋巴结细胞计数及淋巴结细胞中IgE和MCPL-1诱导表达的结果表明蔗糖中不溶性微粒存在引发速发型过敏反应并诱导Th2免疫反应的风险;CCK8法测定不溶性微粒对小鼠脾淋巴细胞增殖的影响结果表明该微粒能直接作用于脾淋巴细胞,提高其增殖能力;而不同厂家的蔗糖产品显示出对小鼠脾淋巴细胞增殖不同程度的影响,作用程度与产品中不溶性微粒的数目有关。以上研究结果提示,当蔗糖用于疫苗注射剂时应关注其中的不溶性微粒。In recent years, adverse reactions caused by vaccines have received much social attention, and the safety of excipients applied in vaccines has also been challenged.Insoluble particles that may be introduced by excipients are one of the risk factors for immune response.At present, although insoluble particles are subject to strict supervision in biological products, there has not been enough research on whether they are derived from excipients as well as the risk contribution brought by excipients.Sucrose is one of the most commonly used protective agents in biotherapeutic formulations in the largest amount, so its safety should be paid enough attention to its quality control.In this study, insoluble particles in sucrose were extracted and prepared by hyperfiltration method, and the particle count, particle size and microscopic morphology were characterized.The particle size extracted from sucrose is characterized from 100 nm to 300 nm;and the insoluble particles of sucrose from different manufacturers are different in size and quantity.BALB/c mice were used to establish an evaluation model for the risk of early immune response activated by insoluble particles in sucrose.The results of toe thickness, lymph node cell count and IgE and MCPL-1 induced expression in lymph node cells showed that insoluble particles in sucrose might induce rapid allergic reaction and induce Th2 immune response.The effect of insoluble particles on the proliferation of mouse spleen lymphocytes was determined by the CCK8 method.It was shown that the particles could directly act on spleen lymphocytes and improve their proliferation ability.The sucrose products from different manufacturers showed different influences on the proliferation of mouse spleen lymphocytes, and the degree of action was related to the number of insoluble particles in the products.The above results suggest that the insoluble particles should be focused when sucrose is used in vaccine injections.

关 键 词：腘窝淋巴结模型 疫苗 辅料 蔗糖 微粒 免疫应答 风险评估 

分 类 号：R965.1[医药卫生—药理学]