title: Dynamic changes of phenotypes and secretory functions during the differentiation of pre-DCs to mature DCs.
abstract: 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 (rmGM-CSF) and 10 ng/mL recombinant mouse interleukin-4 (rmIL-4). On the day 3, 6 and 7 after culture, DCs were divided into non-LPS group and LPS group, given 500 ng/mL 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.
authors.affiliation: Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology; Key Laboratory of the Ministry of Health, Key Laboratory of the Ministry of Education, Wuhan, 430030, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Orthopaedic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Li-Bo
authors.initials: LB
authors.lastname: Zhao
authors.affiliation: Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology; Key Laboratory of the Ministry of Health, Key Laboratory of the Ministry of Education, Wuhan, 430030, China.
authors.forename: Sheng
authors.initials: S
authors.lastname: Chang
authors.affiliation: Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology; Key Laboratory of the Ministry of Health, Key Laboratory of the Ministry of Education, Wuhan, 430030, China.
authors.forename: Chang-Sheng
authors.initials: CS
authors.lastname: Ming
authors.affiliation: Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology; Key Laboratory of the Ministry of Health, Key Laboratory of the Ministry of Education, Wuhan, 430030, China. jyang4@163.com.
authors.forename: Jun
authors.initials: J
authors.lastname: Yang
authors.affiliation: Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology; Key Laboratory of the Ministry of Health, Key Laboratory of the Ministry of Education, Wuhan, 430030, China. nqgong@tjh.tjmu.edu.cn.
authors.forename: Nian-Qiao
authors.initials: NQ
authors.lastname: Gong
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000276:immunology; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry; D016207:Cytokines / Q000378:metabolism*; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology; D005786:Gene Expression Regulation / Q000187:drug effects; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008297:Male; D051379:Mice; D010641:Phenotype
chemical_list: D003470:Culture Media; D016207:Cytokines; D008070:Lipopolysaccharides
keywords: cytokines; dendritic cells; differentiation; lipopolysaccharide