IL-34对类风湿关节炎髓系树突状细胞表型的影响

Effects of IL-34 on phenotype of myeloid dendritic cells in rheumatoid arthritis

目的:探讨IL-34对类风湿关节炎(RA)患者单核细胞向髓系树突状细胞(DC)分化的诱导作用,及其在分化过程中对细胞表型的影响。方法:采集RA患者外周血,Ficoll密度梯度离心法获得PBMC,培养4 h后将贴壁细胞分别用GMCSF+IL-4、IL-4、IL-4+IL-34刺激3 d,流式细胞术检测CD83、CD86和CD14的表达水平;再将GM-CSF+IL-4刺激3 d后的细胞加入TNF-α和(或)IL-34培养4 d,流式细胞术检测CD83、CD86和(或)CD14的表达水平。结果:(1)IL-34+IL-4诱导后CD83、CD86表达水平较IL-4单独作用明显上调(P<0.01),CD14表达无差异(P>0.05);IL-34+IL-4诱导CD86、CD14表达水平较GM-CSF+IL-4刺激组下降(P<0.05),CD83表达无差异(P>0.05)。(2)GM-CSF+IL-4+IL-34诱导CD83、CD86表达水平较GM-CSF+IL-4+TNF-α组低(P<0.05),但与GM-CSF+IL-4刺激组对比CD83、CD86表达水平差异无统计学意义(P>0.05)。(3)GM-CSF+IL-4+TNF-α+IL-34诱导DC CD83表达水平较GM-CSF+IL-4+TNF-α刺激组低(P<0.05),但两组CD86、CD14表达差异无统计学意义(P>0.05)。结论:IL-34在不成熟DC诱导过程中发挥一定作用,效应略弱于GM-CSF;IL-34对成熟DC诱导作用不显著,但参与了不成熟DC的维持。

Objective:To investigate the effect of IL-34 on the phenotype of monocyte derived dendritic cells in RA,and to speculate the role of IL-34 in the differentiation of myeloid dendritic cells.Methods:The peripheral blood of RA patients was collected to harvest PBMC by Ficoll density gradient centrifugation and cultured for 4h.Adherent cells were stimulated with GM-CSF+IL-4,IL-4,IL-4+IL-34 for 3 days,and then the expression of CD83,CD86 and CD14 was tested by flow cytometry.In addition,the cells stimulated by GM-CSF and IL-4 were added by TNF-αwith or without IL-34 for another four days.The expression of CD83,CD86 and/or CD14 was detected by flow cytometry.Results:(1)The expression of CD83 and CD86 on immature DC induced by IL-34+IL-4 was upregulated compared with IL-4 alone(P<0.01),but no difference of the CD14 levels between the two groups(P>0.05).The levels of CD86 and CD14 induced by IL-34+IL-4 were slightly decreased compared with GM-CSF+IL-4 stimulation(P<0.05),but no difference of CD83 expression between the two groups(P>0.05).(2)The expression of CD83 and CD86 stimulated by GM-CSF+IL-4+IL-34 was lower than the GM-CSF+IL-4+TNF-αgroup(P<0.05),but no difference compared with GM-CSF+IL-4 group(P>0.05).(3)The CD83 expression induced by GM-CSF+IL-4+TNF-α+IL-34 was lower than GM-CSF+IL-4+TNF-αgroup(P<0.05),but there was no difference of CD86 and CD14 expression between the two groups(P>0.05).Conclusion:IL-34 played roles in the process of immature DC differentiation,but the effect was slightly weaker than that of GM-CSF.IL-34 did not effect the phenotype change of mature DC,but involved in the maintainence of immature DC.