HL-60分化细胞表面标志、活性及其对肺炎链球菌调理吞噬杀菌能力的动态变化

Dynamic changes of surface markers,cell viability and opsonophagocytic killing capacity of differentiated HL-60 cells against Streptococcus pneumonia

目的分析HL-60分化细胞的表面标志、活性及其对肺炎球菌调理吞噬杀菌能力的动态变化。方法以流式细胞仪连续监测分化1～7 d的HL-60细胞表面标志CD11b、CD35和CD71的表达以及活细胞、凋亡细胞和死亡细胞的比例,同时用09CS、QC2、B、C和F 5份质控血清以调理吞噬杀菌试验检测肺炎链球菌血清型6B、7F、14和23F的杀菌滴度。结果分化3～6 d的HL-60细胞表面标志、活细胞比例可达到实验室要求,5份质控血清的调理吞噬杀菌滴度稳定而且在质控范围之内。结论分化3～6 d的HL-60细胞可以用于评价肺炎链球菌疫苗免疫血清的调理吞噬杀菌试验,为调理吞噬杀菌试验的建立和标准化提供了依据。

Objective After HL-60 cells differentiated for 1 - 7 days, it was to analyze the expression of surface markers, cell viability and the opsonophagocytic killing capacity of these cells against Streptococcus pneumonia. Methods The surface markers CDllb, CD35, CD71 and cell viability of differentiated HL-60 cells were continuously monitored for 1 - 7 days after induction with DMF（ N, N-Dimethyl formamide） by using a flow cytometer. Opsonophagocytic killing assay （OP- KA） were simultaneously performed against pneumococcal serotypes 6B, 7F, 14 and 23F to determine the killing capacity with 5 quality control sera 09CS, QC2, B, C and F, respectively. Results During 3 ~ 6 days after differentiation, the ex- pression of CD11 b, CD35, CD71 and cell viability of differentiated HL60 ceils were in conformity with the essential criteria cited by international laboratories, and the opsonic index （ OI） was stable and fell within the ranges well established for quality controls. Conclusions HL60 cells differentiated on 3 - 6 days can be used in OPKA for evaluation of pneumococcal vaccine, which provide the basis for establishment and standardization of OPKA.