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 stimulati...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.展开更多
基金supported by grants from the National Natural Science Foundation of China(No.81570678,and No.81373169)“973”Major State Basic Research Development Program of China(No.2013CB530803)+1 种基金“863”National High-Tech Researching and Developing Program of Ministry of Science and Technology of China(No.2012AA021010)the Special Project of Ministry of Health of China(No.201302009)
文摘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.
