Dynamic Changes of Phenotypes and Secretory Functions during the Differentiation of Pre-DCs to Mature DCs

• Published in: Journal of Huazhong University of Science and Technology. Medical sciences Vol. 37; no. 2; pp. 191 - 196
• Main Author: 汪晶 赵利波 昌盛 明长生 杨军 宫念樵
• Format: Journal Article
• Language: English
• Published: Wuhan Huazhong University of Science and Technology 01.04.2017; Key Laboratory of the Ministry of Health, Key Laboratory of the Ministry of Education, Wuhan 430030, China%Department of Orthopaedic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China; Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology
• Subjects: Animals; BALB/c小鼠; Bone Marrow Cells - cytology; Bone Marrow Cells - immunology; Cell Differentiation - drug effects; Cells, Cultured; Culture Media - chemistry; Cytokines - metabolism; DCs; Dendritic Cells - cytology; Dendritic Cells - immunology; Gene Expression Regulation - drug effects; Lipopolysaccharides - pharmacology; Male; Medicine; Medicine & Public Health; Mice; Phenotype; 分化过程; 分泌功能; 动态变化; 成熟; 粒细胞-巨噬细胞集落刺激因子; 表型; cytokines; dendritic cells; differentiation; lipopolysaccharide
• ISSN: 1672-0733; 1993-1352; 1993-1352
• DOI: 10.1007/s11596-017-1714-z

The dynamic expression of cytokines and phenotypes during the differentiation process of dendritic cell precursors（pre-DCs） to mature dendritic cells（DCs） was investigated,and the effects of inflammatory stimulation with lipopolysaccharide（LPS） on DCs differentiation were understood.The differentiation of bone marrow cells isolated from Balb/c mice was induced to DCs in an 8-day cell culture system with RPMI-1640 complete culture medium containing 10% FBS,20 ng/mL recombinant mouse granulocyte-macrophage colony-stimulating factor（rm GM-CSF） and 10 ng/m L recombinant mouse interleukin-4（rm IL-4）.On the day 3,6 and 7 after culture,DCs were divided into non-LPS group and LPS group,given 500 ng/m L LPS for 24 h stimulation and no stimulation respectively.The expression levels of CD11c~＋,MHC-Ⅱ~＋,CD40~＋,CD80~＋ and CD86~＋ were detected by flow cytometry,and those of IL-2,IL-4,IL-10,IL-12 p70 and IFN-γ in the supernatant by ELISA.On the day 3 and 6 after culture,the expression of IL-2,IL-4,IL-10 and IFN-γ in DCs showed no significant differences between non-LPS group and LPS group,whereas the differences were significant at day 7.The expression levels of cytokines（for IL-2,IL-4,IL-10,IFN-γ and IL-12 p70：152.86±6.91,778.33±8.35,44.55±2.54,58.26±1.09 and 2423.00±57.21 pg/mL respectively） in LPS group were higher than those in non-LPS group,especially IL-12 p70 increased obviously at day 7.It was concluded that during the differentiation process of pre-DCs to mature DCs,LPS stimulates DCs producing large amounts of IL-12 p70 and Th1-type cytokines as compared with Th2-type cytokines,and day 7 may be a key time-point for DCs polarization.