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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250~300 ml,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3 d,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pDCs活化,减少IFN-α的分泌。
Objective To investigate the effect of hydroxychloroquine (CQQ) on the activation of pDCs induced by CpG-ODN in vitro. Methods The umbilical cord blood was collected from 250 to 300 ml in the delivery room. Umbilical cord blood mononuclear cells were obtained by density gradient centrifugation. Precursor pDCs were sorted from BDCA-4 magnetic beads. Precursor pDCs were divided into three groups (IL-3, IL-3 and CpG , IL-3 + CpG + HCQ) for 3 days. The expressions of BDCA-2, CD86 and CD32 on the surface of pDCs were detected by flow cytometry. The level of IFN-α in supernatant was detected by ELISA. Results ① The expression of BDCA-2 and the fluorescence intensity of IL-3 + CpG group were significantly lower than those of IL-3 group (P <0.05). The expression of CD86 and CD32, the fluorescence intensity and IFN- 3 groups (P <0.05). ② The level of CD32 expression and fluorescence intensity in IL-3 + CpG + HCQ group were significantly lower than that in IL-3 + CpG group (P <0.05); the concentration of IFN-α in culture supernatant was significantly lower than that in IL-3 + CpG group ). Conclusion CpG-ODN can activate pDCs. CD86 and CD32 are highly expressed in activated pDCs and BDCA-2 is low. HCQ inhibits the expression of CD32, inhibits the activation of pDCs induced by CpG-ODN, and decreases the secretion of IFN-α.