β葡聚糖促进小鼠骨髓来源的树突状细胞成熟并增强其迁移能力

β-Glucan promotes the maturation and migration of bone marrow-derived dendritic cells

目的研究β葡聚糖对体外培养小鼠骨髓来源的树突状细胞(BMDC)成熟及迁移能力的影响。方法β葡聚糖体外处理小鼠BMDC,应用流式细胞术检测BMDC表面分子的表达;实时荧光定量PCR(qRT-PCR)检测对照组和β葡聚糖组BMDC中CC趋化因子受体1(CCR1)、CCR2、CCR5、CCR7 mRNA的表达;ELISA检测BMDC上清中白细胞介素6(IL-6)、IL-12p40、肿瘤坏死因子α(TNF-α)和IL-10的水平;TranswellTM迁移实验检测BMDC对CC趋化因子配体19(CCL19)和CCL21的趋化反应性;体内迁移实验检测从皮下迁移至引流淋巴结的DC数目。结果与对照组相比,β葡聚糖能上调BMDC表面分子MHC-Ⅱ类分子、CD80、CD86、CD80、CD40的表达,同时促进BMDC分泌IL-6、TNF-α和IL-12p40,CCR7 mRNA水平增加,增强BMDC对CCL19/CCL21的趋化反应性,并且显著增加从皮下注射部位迁移至引流淋巴结中BMDC数目。结论β葡聚糖不仅能促进BMDC分化成熟,还能增强其体内外迁移能力。

Objective To investigate the effects of β-glucan on the maturation and migration of bone marrow-derived dendritic cells（ BMDCs）. Methods BMDCs were isolated from mouse bone marrow cells in vitro and induced by β-glucan for maturation. The expressions of cell surface markers were detected by flow cytometry（ FCM）. The cytokines（ IL-6,IL-12p40,tumor necrosis factor α） in the supernatants were measured by ELISA,and the expressions of intracellular CC chemokine receptor 1（ CCR1）,CCR2,CCR5,CCR7 were determined by real-time quantitative PCR. Furthermore,the chemotactic response to CC chemokine ligand 19（ CCLl9） and CCL21,i. e. CCR7-1igands,was measured by TranswellTM migration assay. Moreover,the number of migrated cel s in the draining lymph nodes was analyzed by FCM. Results Compared with the control group,the expressions of co-stimulation molecules（ MHC Ⅱ,CD40,CD80,CD86） on BMDCs were up-regulated in the presence of β-glucan. Furthermore,β-glucan could prompt BMDCs to secret high levels of IL-6,TNF-α,IL-12 p40 and increase the production of CCR7 mRNA. After β-glucan treatment,BMDCs were more sensitive to CCL19 /CCL21. The number of BMDCs migrated from subcutaneous injection site into the draining lymph nodes significantly increased in β-glucan group. Conclusion β-glucan can promote the maturation of BMDCs and enhance the migration ability of BMDCs in vitro and in vivo.