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作 者:马斌[1] 汪国生[1] 郭培霞[1] 赵小娟[1] 周晶晶[1] 厉小梅[1] 钱龙[1] 李向培[1]
机构地区:[1]安徽医科大学附属安徽省立医院风湿免疫科,合肥230001
出 处:《免疫学杂志》2013年第1期14-18,共5页Immunological Journal
基 金:国家自然科学基金(81072462);安徽省高校自然科学重点项目(KJ2010A188)
摘 要: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.
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