Xia Qing, Tan Hongyi, Pan Pinhua, Su Xiaoli, Hu Chengping
Department of Respiratory Medicine, Xiangya Hospital, Central South University, Changsha 410008, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2015 Aug;40(8):829-36. doi: 10.11817/j.issn.1672-7347.2015.08.002.
To explore the effect of nerve growth factor (NGF) on the differentiation of murine bone marrow-derived dendritic cells (DCs) in vitro.
The bone marrow cells of femur and tibia from healthy C57B -L/6 mice were isolated and divided into 4 groups: a phosphate buffered saline (PBS) group (PBS group), a NGF group, a granulocyte monocyte colony stimulating factor (GM-CSF) plus interleukin 4 (IL-4) group (GM-CSF+IL-4 group), and a GM-CSF plus IL-4 and NGF group (n=6 in each group). The positive rate of CD11c+ and the proportion of CD8a- were compared at the 7th day among the different groups by flow cytometry. The immature DCs were acquired by classic methods with GM-CSF and IL-4. The purified DCs were obtained by magnetic bead positive selection for CD11c+ cells. The immature DCs were divided into 4 groups: a PBS group, a NGF group, a LPS group, and a NGF+LPS group (n=6 in each group), which were incubated with PBS, NGF, LPS and NGF+LPS, respectively. Cytokine levels of IL-6, IL-10 and IL-12 were detected by ELISA after 24 hours..
NGF can promote the murine bone-marrow cells differentiation into CD11c+ DCs, with CD8a-subset; NGF could enhance LPS-induced cytokine secretion from DCs (IL-6, IL-10 and IL-12).
探讨神经生长因子(NGF)对体外培养的小鼠骨髓来源树突状细胞(DCs)分化的影响。
分离健康C57B -L/6小鼠股骨和胫骨的骨髓细胞,分为4组:磷酸盐缓冲液(PBS)组(PBS组)、NGF组、粒细胞-巨噬细胞集落刺激因子(GM-CSF)加白细胞介素4(IL-4)组(GM-CSF+IL-4组)、GM-CSF加IL-4和NGF组(每组n = 6)。通过流式细胞术比较不同组在第7天时CD11c+阳性率和CD8a-比例。采用经典方法用GM-CSF和IL-4获取未成熟DCs。通过对CD11c+细胞进行磁珠阳性选择获得纯化的DCs。将未成熟DCs分为4组:PBS组、NGF组、脂多糖(LPS)组、NGF+LPS组(每组n = 6),分别用PBS、NGF、LPS和NGF+LPS孵育。24小时后通过酶联免疫吸附测定法(ELISA)检测白细胞介素6(IL-6)、白细胞介素10(IL-10)和白细胞介素12的细胞因子水平。
1)NGF组中CD11c+ DCs的百分比高于PBS组,但低于GM-CSF+IL-4组(均P<0.05)。GM-CSF + IL-4组与NGF+GM-CSF+IL-4组之间无差异(P>0.05)。这四组中CD8a- DCs占主导;2)NGF可进一步上调LPS诱导的DCs细胞因子分泌,如IL-6、IL-10和IL-12(均P<0.05),但单独的NGF无此作用(均P<0.05)。
NGF可促进小鼠骨髓细胞分化为CD11c+ DCs,且为CD8a-亚群;NGF可增强LPS诱导的DCs细胞因子分泌(IL-6、IL-10和IL-12)。