不同来源 HL60细胞用于调理吞噬杀菌试验的比较

摘要

目的：通过比较不同来源的HL60细胞分化后细胞的表面标志、活性及其对肺炎链球菌调理吞噬杀菌指数的动态变化，评价HL60细胞的来源对肺炎链球菌调理吞噬杀菌能力的影响。方法使用流式细胞仪连续监测来源于美国ATCC和中国科学院上海细胞研究所的HL60细胞在分化后1～7 d的表面标志CD11b、CD35和CD71的表达情况，并观察活细胞、凋亡细胞和死亡细胞的比例；同时使用两种不同来源的细胞检测09 CS和B 两份质控血清对肺炎链球菌血清型6B、7F、14和23F菌株的调理吞噬杀菌指数，同时观察09CS对13价结合疫苗血清型1、3、4、5、6A、6B、7F、9V1、4、18C、19A、19F和23F菌株的调理吞噬杀菌指数。结果两种不同来源HL60细胞分化3～6d细胞的表面标志、活细胞比例均可达到国际实验室的要求指标；两份质控血清对检测菌株的调理吞噬杀菌指数均能稳定于质控范围之内；两种细胞检测的09 CS对13价肺炎链球菌结合疫苗血清型菌株的调理吞噬杀菌指数具可比性。结论两种来源不同的 HL60细胞分化3～6 d均能用于评价肺炎链球菌疫苗免疫血清的调理吞噬杀菌试验，这为调理吞噬杀菌试验及其标准化在国内的建立提供了便利。%Objective To assess the influence of HL60 cells purchased from different sources on the opsonophagocytic kill-ing capacity against Streptococcus pneumonia by comparison of the expression of surface markers , cell viability and opsonic index.Methods The cell surface markers of CD11b, CD35, CD71 and cell viability of differentiated HL60 cells were daily monitored for 1-7 days after induction with DMF ( N, N-Dimethyl formamide ) by using flow cytometer .Opsonophagocytic killing assay ( OPKA) were simultaneously performed against pneumococcal strains of serotype 6B, 7F, 14 and 23F with 2 quality control sera 09CS and B as well as against strains of 13 valent conjugate vaccine serotype 1,3,4,5,6A, 6B, 7F, 9V,14, 18C, 19A, 19F and 23F with 09CS, by using differentiated HL60 cell from different sources .Results During 3-6 days after differentiation, irrespective of HL60 cell sources,the expression of CD11b, CD35, CD71 and cell viability of differentiated HL60 cells met the essential criteria cited among international laboratories , and the opsonic indexes ( OI) tested for given strains were stable and fell within the ranges well established for quality controls .Conclusions HL60 cells from both ATCC and Shanghai can be used in OPKA for evaluation of pneumococcal vaccine ,which will facilitate the estab-lishment and standardization of OPKA in domestic .