羟氯喹对CpG-ODN诱导浆细胞样树突状细胞活化影响的初步研究

Impact of hydroxychloroquine on pl CpG-ODN-induced asmacytoid dendritic cells activation

目的体外探讨羟氯喹（HCQ）对CpG—ODN诱导pDCs活化的影响。方法产房收集脐带血250-300ml．密度梯度离心得脐带血单核细胞，BDCA-4磁珠分选得前体pDCs细胞，前体pDCs分为3组（IL-3，IL-3＋CpG，IL-3＋CpG＋HCQ）培养3d．流式细胞术测pDCs表面BDCA-2、CD86、CD32表达，ELISA检测上清细胞因子IFN-α水平。结果①IL-3＋CpG组BDCA-2表达水平和荧光强度均显著低于IL-3组（P〈0．05）；CD86和CD32表达水平、荧光强度和培养上清IFN—α浓度显著高于IL-3组（P〈0．05）。②IL-3＋CpG＋HCQ组CD32表达水平和荧光强度均显著低于IL-3＋CpG组（P〈0．05）；培养上清IFN—α浓度显著低于IL-3＋CpG组（P〈0．05）。结论CpG—ODN可以活化pDCs，活化的pDCs高表达CD86、CD32，低表达BDCA-2；HCQ抑制CD32表达．抑制CpG—ODN诱导的DDCs活化．减少IFN-α的分泌。

To investigate the impact of hydroxychloroquine （HCQ） on plasmacytoid dendritic cells activation induced by CpG-ODN in vitro, cord blood （250-300 ml） was collected from delivery room. Density gradient centrifugation was used to get the cord blood monocytes and BDCA-4 beads sorting was used to obtain the precursor pDCs. Precursor pDCs was co-cuhured with IL-3, IL-3 plus CpG-ODN2216, and IL-3 plus CpG- ODN2216＋HCQ, respectively for 3 days. Then flow cytometry was used to measure the pDCs surface expression of BDCA-2, CD86 and CD32; while ELISA was employed to detect the concentration of IFN-α in the supernatants. We found that CpG-ODN2216 activated pDCs demonstrated higher expressions of CD86 and CD32, more IFN-α production, but lower expression of BDCA-2 in vitro （P 〈 0.05）. And HCQ could significantly down-regulate the CD32 expression, CD32 MFI level, and IFN-α level in the supernatants （P〈 0.05）. In conclusion, HCQ may inhibit pDCs activation and IFN-α secretion induced by CpG-ODN2216 via reducing CD32 expression on pDCs.