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目的:探讨加入LPS、TGF-β1体外刺激小鼠骨髓来源树突状细胞(BMDC)的方法及其对生物学特性的作用。方法:GM-CSF和IL-4诱导培养小鼠BMDC6d后,分别用培养基(对照组)、LPS、TGF-β1、LPS+TGF-β1,刺激BMDC48h后进行形态学观察,流式细胞术检测细胞表型CD11C、CD80、CD86、MHCⅡ,混合淋巴细胞反应检测其抗原提呈功能,收集上清液用ELISA检测IL-6,IL-12p70。结果:LPS组具有最典型的成熟样DC形态,CD80、CD86及MHCⅡ的表达水平显著升高,混合淋巴细胞反应和分泌IL-4、IL-12p70能力最强,与对照组,TGF-β1组,LPS+TGF-β1组比较差异具有统计学意义(P<0.05),TGF-β1组成熟样DC形态最不典型,CD80、CD86和MHCⅡ的表达水平最低,混合淋巴细胞反应和分泌IL-4、IL-12p70能力最弱,与对照组,LPS组比较差异具有统计学意义(P<0.05)。结论:LPS在DC的分化晚期可以刺激其成熟,并且具有更高的生物学特性,TGF-β1不抑制DC的分化,但可以抑制DC的成熟,从而降低其生物学特性。
Objective: To investigate the method of stimulating mouse bone marrow-derived dendritic cells (BMDC) stimulated by LPS and TGF-β1 in vitro and its effect on biological characteristics. Methods: After BMDC6d induced by GM-CSF and IL-4, the morphological changes of BMDC were observed by using culture medium (control group), LPS, TGF-β1, LPS and TGF- Cell phenotype CD11C, CD80, CD86, MHC Ⅱ, mixed lymphocyte reaction to detect antigen presenting function, the supernatant was collected to detect IL-6, IL-12p70 by ELISA. Results: The LPS group had the most typical mature DC morphology, the expression levels of CD80, CD86 and MHC Ⅱ were significantly increased, and the ability of mixed lymphocyte reaction and secretion of IL-4 and IL-12p70 was the strongest. Compared with the control group and TGF-β1 group , LPS + TGF-β1 group (P <0.05). The DC-like morphology of TGF-β1 group was the most atypical, CD80, CD86 and MHCⅡwere the lowest, mixed lymphocyte reaction and secretion of IL-4 , IL-12p70 was the weakest, compared with the control group and LPS group, the difference was statistically significant (P <0.05). CONCLUSION: LPS can stimulate its maturation in the later stage of DC differentiation and has higher biological characteristics. TGF-β1 does not inhibit the differentiation of DC, but it can inhibit the maturation of DC and decrease its biological characteristics.