HBeAg对小鼠骨髓源性树突状细胞成熟的影响  

Effects of hepatitis B e antigen on the maturation of murine bone marrow-derived dendritic cells

作　　者：蓝松松 吴乐灿 吴金明[1] 林贤凡[1] 吴文治[1] 王秀燕[1] 黄智铭[1] 吴建胜[1] 

机构地区：[1]温州医科大学附属第一医院消化内科,325000

出　　处：《免疫学杂志》2015年第2期116-120,共5页Immunological Journal

基　　金：浙江省自然科学基金(LY12H03003;Y2110768)

摘　　要：目的探讨HBe Ag对LPS诱导小鼠骨髓源性树突状细胞(DC)成熟的影响。方法体外诱导C57BL/6小鼠骨髓细胞分化成未成熟树突状细胞,经CD11c磁珠分选纯化后将DCs随机分为空白对照组、LPS刺激组、HBe Ag+LPS刺激组。流式检测DC表型变化,混合淋巴反应(MLR)检测DC促T淋巴细胞增殖能力,酶联免疫法(ELISA)检测细胞上清液中IL-12的分泌水平,Western blot检测p38磷酸化水平,并设置SB203580组为阳性对照探讨细胞IL-12分泌的可能调节机制。结果 LPS刺激未成熟DC引起细胞表面MHC-Ⅱ、CD86表达升高,刺激同种异体淋巴细胞增殖能力增强,IL-12分泌量增高。HBe Ag可抑制LPS促进DC表面MHC-Ⅱ、CD86表达升高和促淋巴细胞增殖能力增强的作用。LPS刺激DC可引起p38磷酸化水平升高,并呈时间依赖性;HBe Ag或SB203580预处理细胞再予LPS刺激,磷酸化p38表达和IL-12分泌较单纯LPS刺激组明显下降。结论 HBe Ag对LPS引起的树突状细胞的成熟有一定的抑制作用,且HBe Ag可能通过抑制p38MAPK信号通路下调LPS诱导的树突状细胞IL-12的产生。The study performed to investigate the effects of HBe Ag on the maturation of lipopolysaccharide（LPS） stimulated murine bone marrow-derived dendritic cells（BM-DCs）. The murine bone marrow cells were cultured and induced into DCs in vitro. Then the DCs were purified by murine CD11 c microbeads,and divided into control group,LPS-stimulated group,LPS plus HBe Ag-treated group. Flow cytometry was used to analyze the expression of CD86 and MHC- Ⅱ; the mixture lymphocyte reaction was tested by cell counting kit-8; IL-12 production was assessed by ELISA; the induced changes in expression of phospho-p38（p-p38） were measured by Western blotting. Additionally,SB203580 was used as a positive group to expore the potential mechanism of IL-12 regulation. The data showed that LPS induced higher expression levels of the activation markers CD86 and MHC-Ⅱ,and an enhanced capacity to stimulate allogeneic T cell proliferation compared with the control group and HBe Ag plus LPS group. Meanwhile,DCs treated with LPS showed an increased production of IL-12 and an up-regulated expression of intracellular p-p38 protein. While HBe Ag and SB203580 all led to significant reduction of p-p38 expression compared with LPS-stimulated group. LPS could significantly promote IL-12 secretion,but there was no obvious increase of IL-12 level in the cells pretreated with HBe Ag or SB203580. Our findings indicated that HBe Ag may play a negative role in the activation of LPS-stimulated DCs. p38 MAPK signaling pathway contributes to the production of IL-12,and HBe Ag may suppress the secretion of IL-12 by inhibiting the phosphorylation of p38.

关 键 词：肝炎e抗原乙型 树突状细胞 脂多糖 P38丝裂原活化蛋白激酶类 白细胞介素-12 

分 类 号：R512.62[医药卫生—内科学]