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-gamma in the supernatant by ELISA. On the day 3 and 6 after culture, the expression of IL-2, IL-4, IL-10 and IFN-gamma 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-gamma 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.
基金:
National Natural Science Foundation of ChinaNational Natural Science Foundation of China (NSFC) [81570678, 81373169]; "973" Major State Basic Research Development Program of ChinaNational Basic Research Program of China [2013CB530803]; "863" National High-Tech Researching and Developing Program of Ministry of Science and Technology of China [2012AA021010]; Ministry of Health of China [201302009]
第一作者单位:[1]Huazhong Univ Sci & Technol,Inst Organ Transplantat,Tongji Hosp,Tongji Med Coll,Wuhan 430030,Peoples R China
通讯作者:
通讯机构:[1]Huazhong Univ Sci & Technol,Inst Organ Transplantat,Tongji Hosp,Tongji Med Coll,Wuhan 430030,Peoples R China[2]Minist Educ, Key Lab, Minist Hlth, Wuhan 430030, Peoples R China
推荐引用方式(GB/T 7714):
wang jing,zhao li-bo,chang sheng,et al.Dynamic changes of phenotypes and secretory functions during the differentiation of pre-DCs to mature DCs[J].JOURNAL OF HUAZHONG UNIVERSITY OF SCIENCE AND TECHNOLOGY-MEDICAL SCIENCES.2017,37(2):191-196.doi:10.1007/s11596-017-1714-z.
APA:
wang,jing,zhao,li-bo,chang,sheng,ming,chang-sheng,yang,jun&gong,nian-qiao.(2017).Dynamic changes of phenotypes and secretory functions during the differentiation of pre-DCs to mature DCs.JOURNAL OF HUAZHONG UNIVERSITY OF SCIENCE AND TECHNOLOGY-MEDICAL SCIENCES,37,(2)
MLA:
wang,jing,et al."Dynamic changes of phenotypes and secretory functions during the differentiation of pre-DCs to mature DCs".JOURNAL OF HUAZHONG UNIVERSITY OF SCIENCE AND TECHNOLOGY-MEDICAL SCIENCES 37..2(2017):191-196