热休克蛋白65-MUC1抗原肽融合蛋白效力测定方法的建立

Development of A Method for Determination of Potency of Heat Shock Protein 65-MUC1 Antigen Peptide Fusion Protein

目的建立热休克蛋白65(HSP65)MUC1抗原肽(HSP65MUC1)融合蛋白的效力测定方法。方法从人外周血中分离出单核细胞,并用人IL4和GMCSF将其诱导为未成熟的树突状细胞(iDCs),将HSP65MUC1融合蛋白加载至iDC中继续培养2d后,用流式细胞仪检测树突状细胞(DCs)表面CD80、CD83和CD86等的表达。结果HSP65MUC1融合蛋白能使iDCs表面的CD80、CD83和CD86等表达上调,其中CD86的表达上调程度与其加载HSP65MUC1融合蛋白的量成正相关。结论通过检测对人iDCs表面CD86表达的刺激作用来检测HSP65MUC1融合蛋白效力的实验方法具有操作简便、周期短和重复性好等优点。

Objective To develop a method for determining the potency of heat shock protein 65-MUC1 antigen peptide fusion protein(HSP65-MUC1).Methods Immature dendritic cells(iDCs) were derived from human PBMC by induction with human IL-4 and GM-CSF.Add HSP65-MUC1 fusion protein into iDC and incubate for 2 days.Determine the expressions of CD80,CD83 and CD86 on the surface of DCs by flow cytometer.Results HSP65-MUC1 fusion protein enhanced the expressions of CD80,CD83 and CD86.The up-regulation of CD86 was positively related to the amount of HSP65-MUC1 added into iDCs.Conclusion A method for determining the potency of HSP65-MUC1 fusion protein by testing the expression of CD86 on the surface of iDCs was developed.The method was simple,time-saving and of good reproducibility.