蚌埠医学院学报
蚌埠醫學院學報
방부의학원학보
ACTA ACADEMIAE MEDICINAE BENGBU
2015年
9期
1145-1147,1151
,共4页
金齐力%孙悝%吕小艳%韦莉
金齊力%孫悝%呂小豔%韋莉
금제력%손리%려소염%위리
结核分枝杆菌%树突状细胞%免疫应答%小鼠
結覈分枝桿菌%樹突狀細胞%免疫應答%小鼠
결핵분지간균%수돌상세포%면역응답%소서
mycobacterium tuberculosis%dendritic cells%immune response%mouse
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。
目的::探討結覈分枝桿菌( Mycobacterium tuberculosis,Mtb)感染對小鼠骨髓來源的樹突狀細胞( DC)功能的影響。方法:將DC分為4組,即脂多糖( LPS)感染組、Mtb感染組、滅活Mtb感染組和正常細胞對照組。以LPS、Mtb及滅活的Mtb與小鼠骨髓來源的DC建立小鼠體外感染模型,用酶聯免疫吸附法測定白細胞介素( IL)-6、IL-12及腫瘤壞死因子-α的錶達;流式細胞術檢測DC錶麵主要組織相容性複閤體Ⅱ類分子、CD40、CD80和CD86的錶達。結果:每隻小鼠的股骨骨髓可擴增穫得5×106~1×107箇具有典型細胞形態的DC,純度達85%以上;與對照組比較,流式細胞術結果顯示LPS、Mtb和滅活Mtb感染組均能促進DC錶麵分子的錶達(P<0.01)。 Mtb感染組DC錶麵分子上調顯著低于LPS感染組與滅活Mtb感染組(P<0.01);LPS、Mtb或滅活Mtb作用後,DC的IL-6、IL-12和腫瘤壞死因子-α分泌量顯著增加(P<0.01),Mtb感染組DC的細胞因子分泌量顯著低于LPS感染組與滅活Mtb感染組(P<0.01)。結論:Mtb活菌可榦擾DC的細胞因子分泌,抑製DC成熟,從而削弱其抗原遞呈的功能,影響抗原特異性細胞的免疫活化。
목적::탐토결핵분지간균( Mycobacterium tuberculosis,Mtb)감염대소서골수래원적수돌상세포( DC)공능적영향。방법:장DC분위4조,즉지다당( LPS)감염조、Mtb감염조、멸활Mtb감염조화정상세포대조조。이LPS、Mtb급멸활적Mtb여소서골수래원적DC건립소서체외감염모형,용매련면역흡부법측정백세포개소( IL)-6、IL-12급종류배사인자-α적표체;류식세포술검측DC표면주요조직상용성복합체Ⅱ류분자、CD40、CD80화CD86적표체。결과:매지소서적고골골수가확증획득5×106~1×107개구유전형세포형태적DC,순도체85%이상;여대조조비교,류식세포술결과현시LPS、Mtb화멸활Mtb감염조균능촉진DC표면분자적표체(P<0.01)。 Mtb감염조DC표면분자상조현저저우LPS감염조여멸활Mtb감염조(P<0.01);LPS、Mtb혹멸활Mtb작용후,DC적IL-6、IL-12화종류배사인자-α분비량현저증가(P<0.01),Mtb감염조DC적세포인자분비량현저저우LPS감염조여멸활Mtb감염조(P<0.01)。결론:Mtb활균가간우DC적세포인자분비,억제DC성숙,종이삭약기항원체정적공능,영향항원특이성세포적면역활화。
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 classⅡ,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.