结核分枝杆菌感染对小鼠树突状细胞功能的影响

    Effects of Mycobacterium tuberculosis on dendritic cells function

    • 摘要: 目的:探讨结核分枝杆菌(Mycobacterium tuberculosis,Mtb)感染对小鼠骨髓来源的树突状细胞(DC)功能的影响。方法:将DC分为4组,即脂多糖(LPS)感染组、Mtb感染组、灭活Mtb感染组和正常细胞对照组。以LPS、Mtb及灭活的Mtb与小鼠骨髓来源的DC建立小鼠体外感染模型,用酶联免疫吸附法测定白细胞介素(IL)-6、IL-12及肿瘤坏死因子-α的表达;流式细胞术检测DC表面主要组织相容性复合体Ⅱ类分子、CD40、CD80和CD86的表达。结果:每只小鼠的股骨骨髓可扩增获得5×106~1×107个具有典型细胞形态的DC,纯度达85%以上;与对照组比较,流式细胞术结果显示LPS、Mtb和灭活Mtb感染组均能促进DC表面分子的表达(P<0.01)。Mtb感染组DC表面分子上调显著低于LPS感染组与灭活Mtb感染组(P<0.01);LPS、Mtb或灭活Mtb作用后,DC的IL-6、IL-12和肿瘤坏死因子-α分泌量显著增加(P<0.01),Mtb感染组DC的细胞因子分泌量显著低于LPS感染组与灭活Mtb感染组(P<0.01)。结论:Mtb活菌可干扰DC的细胞因子分泌,抑制DC成熟,从而削弱其抗原递呈的功能,影响抗原特异性细胞的免疫活化。

       

      Abstract: Objective:To investigate the effect of Mycobacterium tuberculosis(Mtb) on the function of mouse bone marrow derived dendritic cells(DC).Methods:DC were divided into 4 groups,lipopolysaccharide(LPS)-treated group,Mtb-treated group,inactivated Mtb-treated group and control group.Cytokines including interleukin(IL)-6,IL-12 and tumor necrosis factor α secretion of infected cells were examined by ELISA method.And the expression of surface molecules of infected DC including major histocompatibility complex class Ⅱ,CD40,CD80 and CD86 were detected by flow cytometry.Results:About 5×106 to 1×107 DC were obtained from bone marrow of one mouse,and the purity of DC was more than 85%.The cells had the typical morphology of DC.The expressions of major histocompatibility complex class Ⅱ,CD40,CD80 and CD86 on stimulated DC increased in LPS-treated group,Mtb-treated group,inactivated Mtb-treated group in contrast to control group;while the percentage of the upregulation of surface molecules in Mtb-treated DC was significantly lower than that of LPS or inactivated Mtb-treated DC.The cytokine levels in all groups significantly increased,and IL-6,IL-12 and tumor necrosis factor α secretions of DC infected by Mtb were significantly lower than those of LPS or inactivated Mtb-treated DC.Conclusions:Mtb can decrease the antigen presenting function through interfering the secretion of cytokines and inhibiting maturation of DC,and consequently suppress DC and T cell activation and result in the more serious damage of host.

       

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