结核分枝杆菌感染对小鼠树突状细胞功能的影响

金齐力; 孙悝; 吕小艳; 韦莉

doi:10.13898/j.cnki.issn.1000-2200.2015.09.001

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结核分枝杆菌感染对小鼠树突状细胞功能的影响

金齐力 , 孙悝 , 吕小艳 , 韦莉

文章导航 > 蚌埠医学院学报 > 2015, 40 > 9: (1145-1147,1151)

引用本文:

Citation:

结核分枝杆菌感染对小鼠树突状细胞功能的影响

1. 蚌埠医学院第二附属医院检验科, 安徽蚌埠 233040; 蚌埠医学院;
2. 感染与免疫安徽省重点实验室;
3. 病原生物学教研室, 安徽蚌埠 233030

作者简介: 金齐力(1979-),男,硕士,主管检验师,讲师

通讯作者: 韦莉, 副教授.weili060903@126.com

基金项目:
安徽省自然科学基金项目(1508085QH175)

安徽省高校省级自然科学研究项目(KJ2013B142)
中图分类号: R378.911

Effects of Mycobacterium tuberculosis on dendritic cells function

1. Department of Clinical Laboratory, The Second Affiliated Hospital of Bengbu Medical College, Bengbu Anhui 233040;
2. Anhui Key Laboratory of Infection and Immunity;
3. Department of Microbiology and Parasitology, Bengbu Medical College, Bengbu Anhui 233030, China

Corresponding author: WEI Li, 副教授.weili060903@126.com

CLC number: R378.911

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。
- 结核分枝杆菌 /
- 树突状细胞 /
- 免疫应答 /
- 小鼠
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.
- mycobacterium tuberculosis /
- dendritic cells /
- immune response /
- mouse

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结核分枝杆菌感染对小鼠树突状细胞功能的影响

通讯作者: 韦莉, 副教授.weili060903@126.com

作者简介: 金齐力(1979-),男,硕士,主管检验师,讲师

1. 蚌埠医学院第二附属医院检验科, 安徽蚌埠 233040; 蚌埠医学院;
2. 感染与免疫安徽省重点实验室;
3. 病原生物学教研室, 安徽蚌埠 233030

收稿日期: 2014-09-02

基金项目: 安徽省自然科学基金项目(1508085QH175)安徽省高校省级自然科学研究项目(KJ2013B142)

关键词:

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。

English Abstract

Effects of Mycobacterium tuberculosis on dendritic cells function

Corresponding author: WEI Li, 副教授.weili060903@126.com

1. Department of Clinical Laboratory, The Second Affiliated Hospital of Bengbu Medical College, Bengbu Anhui 233040;
2. Anhui Key Laboratory of Infection and Immunity;
3. Department of Microbiology and Parasitology, Bengbu Medical College, Bengbu Anhui 233030, China

Received Date: 2014-09-02

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.