英夫利昔单抗对小鼠骨髓树突状细胞的免疫调节作用及其机制探讨

Immunomodulatory effects of Infliximab on maturation and migration of bone marrow derived dendritic cells

目的探讨英夫利昔单抗(Infliximab)对小鼠骨髓树突状细胞(BMDCs)的免疫调节作用及机制。方法分离6-8周健康C57BL/6小鼠骨髓,刺激诱导产生未成熟DCs,分别采用0、10μg/ml、50μg/ml、250μg/ml的Infliximab处理BMDCs,检测其凋亡坏死情况。进一步用脂多糖(LPS)刺激,检测协同共刺激分子CD40、CD80、CD86和抗原递呈分子MHCⅡ类抗原表达情况;Infliximab预处理BMDCs,再用LPS刺激,用迁移小室进行培养,用CCL21对BMDCs进行趋化,检测BMDCs迁移能力的变化。结果 250μg/ml的Infliximab作用后BMDCs的凋亡坏死明显增加,因此实验剂量选用50μg/ml;Infliximab处理的BMDCs在LPS刺激后,CD40、CD80、CD86及MHCⅡ类抗原表达下调;Infliximab处理的BMDCs在LPS刺激后对CCL21的趋化反应性下降(P均<0.05)。结论英夫利昔单抗(Infliximab)通过影响小鼠骨髓树突状细胞(BMDCs)的成熟、迁移来抑制BMDCs对适应性免疫反应的启动作用,进而影响自身免疫性疾病的发生发展。

Objective To investigate the effect of Infliximab on maturation and migration of murine bone marrowderived dendritic cells(BMDCs)and its potential mechanism.Methods BMDCs were generated in vitro from bone marrow of C57BL/6mice by being cultured with GM-CSF.For cytotoxicity assay,BMDCs were treated with different concentrations of Infliximab(0,10μg/ml,50μg/ml and 250μg/ml)for 24 h,the rate of apoptosis and necrosis was detected by flow cytometry.For maturation and migration analysis,BMDCs were treated with LPS(1μg/ml)in the presence of Infliximab.And the expression of CD40、CD80、CD86and MHCII on BMDCs and the ability of migaration were analyzed by using Flowcytometry.Results Infliximab induced significant BMDC apoptosis and necrosis only at concentrations of 250ug/ml or more(P<0.05),but not at the concentrations of 50μM or less.Infliximab markedly suppressed the expression of costimulatory molecules,such as CD40,CD80,CD86,and MHC class II on the surfaces of the LPSstimulated DCs.(P<0.05).And aslo Infliximab inhibited the migration of LPS-treated BMDCs in vitro ConclusionInfliximab could suppress the maturation and migration of mouse BMDCs in vitro culture.