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单核细胞增生李斯特菌L型对小鼠脾脏树突状细胞免疫调节作用影响的研究

Research on Immunomodulatory Effects of Listeria monocytogenes L-form on Mouse Splenic Dendritic Cells

  • 摘要:
      目的  研究单核细胞增生李斯特菌(Listeria monocytogenes, LM)感染对树突状细胞(dendritic cell, DC)免疫功能的影响,探讨细菌型(WT)和L型(L-form)LM对DC的免疫活化能力差异。
      方法  将C57BL/6小鼠随机分为对照组、WT组、L-form组,3组小鼠分别经尾静脉感染PBS液、细菌型LM和L型LM,电镜观察脾脏DC超微结构特征;流式细胞术检测小鼠脾脏DC的数量、共刺激分子表达、胞内细胞因子分泌、脾脏T细胞亚群及其活化程度。
      结果  透射电镜可观察到对照组小鼠脾脏DC表面有大量丝状伪足,胞浆均匀,细胞核大而偏于一侧;吞噬细菌后,表面丝状伪足减少,胞质内空泡增多;小鼠感染后第1天脾脏中DC绝对值无明显升高或降低(P>0.05),但成熟表型特征性分子表达上调(P < 0.05),L-form感染组DC表面CD80和CD86分子均高于WT组(P < 0.05);小鼠感染LM后TNF-α+DC比例明显升高,L-form感染组分泌TNF-α的DC高于WT组(P < 0.05);感染LM后第7天,3组小鼠脾脏CD4+T细胞和CD8+T细胞在淋巴细胞中所占比例差异均无统计学意义(P>0.05),但L-form组CD69+ T细胞比例高于WT组(P < 0.05)。
      结论  L型LM可介导相对高水平的TNF-α,促进DC成熟以增强其抗原递呈的能力。

     

    Abstract:
      Objective  To study the effect of Listeria monocytogenes (LM) infection on dendritic cell (DC) immune function, and to compare the difference of immune activation ability on DC cell between bacterial form (WT) and L-form.
      Methods  C57BL/6 mice were randomly divided into 3 groups and infected intravenously with LM WT, LM L-form or PBS respectively. The ultrastructural characteristics of splenic DC were observed by transmission electron microscopy (TEM). The number of splenic DC, the expression of costimulatory molecules, the secretion of cytokine and the activation of splenic T lymphocyte were detected by flow cytometry.
      Results  TEM observation found that there were a large number of filamentous pseudopodia on the surface of splenic DC cells. The cytoplasm of DC was homogeneous and its nucleus was large. After phagocytosis of bacteria, the number of pseudopodia on the surface decreased and vacuoles in the cytoplasm increased. The number of splenic DC did not show significant changes at 1 d post infection (P>0.05). However, the expression of mature phenotypic molecules were significantly up-regulated (P < 0.05), in L-form infection group, both CD80 and CD86 molecules expressed on DC surface were higher than WT group (P < 0.05). Compared with control group, the proportion of TNF-α+DC were elevated after LM infection, and the average percentage of TNF-α+DC of L-form infection group was significantly higher than that of WT group (P < 0.05). At 7 d after infection, the average percentage of CD69+ T cells of L-form group was significantly higher than that of WT group (P < 0.05).
      Conclusion  LM L-form can induce relatively high levels of TNF-α and promote DC maturation so that to enhance their antigen presenting ability.

     

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