中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2008年
48期
3392-3396
,共5页
鼻炎%变应性%辅助性T细胞%螨%T细胞免疫球蚩白功能区和黏液素样功能区-1(TIM-1)
鼻炎%變應性%輔助性T細胞%螨%T細胞免疫毬蚩白功能區和黏液素樣功能區-1(TIM-1)
비염%변응성%보조성T세포%만%T세포면역구치백공능구화점액소양공능구-1(TIM-1)
Rhinitis,allergic%Helper T lymphocytes%Dermatophagoides pteryonyssinus%Tcell immunoglobulin domain and mucin domain-a (TIM-1)
目的 探讨人辅助性T细胞(helperT lymphocyte,Th)表达T细胞免疫球蛋白功能区和黏液素样功能区-1(T cell immunoglobulin domain and mucin domain-1,TIM-1)在变应性鼻炎(allergicrhinitis,AR)中的作用.方法 密度梯度离心分离20名健康人和20例AR患者外周血单个核细胞(human peripheral blood mononuclear cells,PBMC),免疫磁珠分选CD4+T细胞,CD4+CD45RA+T细胞,免疫荧光和流式细胞仪检测TIM-1阳性细胞.对比植物血凝素(phytohemagglutinin,PHA,20μg/ml)非特异性刺激上述细胞后表达TIM-1阳性细胞的差异.标准变应原(屋尘螨,50000 SOU/ml)和内毒素(脂多糖,lipopolysaccharide,LPS,20ng/ml)特异性刺激树突状细胞(dendritic cells,DC),观察DC提呈抗原并诱导初始Th细胞表达TIM-1的特点.酶联免疫吸附实验(ELISA)检测培养液中IL-4、IFN-γ的分泌.结果 AR患者组CD4+T细胞中TIM-1阳性CD4+T细胞9.48%4±1.51%,高于健康组3.12%±0.32%(P＜0.05);PHA刺激后两组表达TIM-1的CD4+T细胞均高于刺激前(P值均<0.05).DC提呈抗原刺激健康组初始Th细胞后,检测混合细胞液中表达TIM-1的细胞,标准变应原组10.81%±1.48%,高于LPS组6.07%±1.45%(P＜0.05).ELISA检测相应IL-4、IFN-γ的分泌,标准变应原组IL-4为(74.61±13.82)pg/ml、(25.31±7.23)pg/ml;LPS组分别为(28.57±3.36)pg/ml、(163.41±82.37)Pg/ml.结论 AR患者中表达TIM-1的Th细胞高于健康者,DC提呈抗原后可以促进其表达;TIM-1的表达有利于初始Th细胞向Th2分化,由此诱发以Th2细胞因子为主的免疫反应.
目的 探討人輔助性T細胞(helperT lymphocyte,Th)錶達T細胞免疫毬蛋白功能區和黏液素樣功能區-1(T cell immunoglobulin domain and mucin domain-1,TIM-1)在變應性鼻炎(allergicrhinitis,AR)中的作用.方法 密度梯度離心分離20名健康人和20例AR患者外週血單箇覈細胞(human peripheral blood mononuclear cells,PBMC),免疫磁珠分選CD4+T細胞,CD4+CD45RA+T細胞,免疫熒光和流式細胞儀檢測TIM-1暘性細胞.對比植物血凝素(phytohemagglutinin,PHA,20μg/ml)非特異性刺激上述細胞後錶達TIM-1暘性細胞的差異.標準變應原(屋塵螨,50000 SOU/ml)和內毒素(脂多糖,lipopolysaccharide,LPS,20ng/ml)特異性刺激樹突狀細胞(dendritic cells,DC),觀察DC提呈抗原併誘導初始Th細胞錶達TIM-1的特點.酶聯免疫吸附實驗(ELISA)檢測培養液中IL-4、IFN-γ的分泌.結果 AR患者組CD4+T細胞中TIM-1暘性CD4+T細胞9.48%4±1.51%,高于健康組3.12%±0.32%(P＜0.05);PHA刺激後兩組錶達TIM-1的CD4+T細胞均高于刺激前(P值均<0.05).DC提呈抗原刺激健康組初始Th細胞後,檢測混閤細胞液中錶達TIM-1的細胞,標準變應原組10.81%±1.48%,高于LPS組6.07%±1.45%(P＜0.05).ELISA檢測相應IL-4、IFN-γ的分泌,標準變應原組IL-4為(74.61±13.82)pg/ml、(25.31±7.23)pg/ml;LPS組分彆為(28.57±3.36)pg/ml、(163.41±82.37)Pg/ml.結論 AR患者中錶達TIM-1的Th細胞高于健康者,DC提呈抗原後可以促進其錶達;TIM-1的錶達有利于初始Th細胞嚮Th2分化,由此誘髮以Th2細胞因子為主的免疫反應.
목적 탐토인보조성T세포(helperT lymphocyte,Th)표체T세포면역구단백공능구화점액소양공능구-1(T cell immunoglobulin domain and mucin domain-1,TIM-1)재변응성비염(allergicrhinitis,AR)중적작용.방법 밀도제도리심분리20명건강인화20례AR환자외주혈단개핵세포(human peripheral blood mononuclear cells,PBMC),면역자주분선CD4+T세포,CD4+CD45RA+T세포,면역형광화류식세포의검측TIM-1양성세포.대비식물혈응소(phytohemagglutinin,PHA,20μg/ml)비특이성자격상술세포후표체TIM-1양성세포적차이.표준변응원(옥진만,50000 SOU/ml)화내독소(지다당,lipopolysaccharide,LPS,20ng/ml)특이성자격수돌상세포(dendritic cells,DC),관찰DC제정항원병유도초시Th세포표체TIM-1적특점.매련면역흡부실험(ELISA)검측배양액중IL-4、IFN-γ적분비.결과 AR환자조CD4+T세포중TIM-1양성CD4+T세포9.48%4±1.51%,고우건강조3.12%±0.32%(P＜0.05);PHA자격후량조표체TIM-1적CD4+T세포균고우자격전(P치균<0.05).DC제정항원자격건강조초시Th세포후,검측혼합세포액중표체TIM-1적세포,표준변응원조10.81%±1.48%,고우LPS조6.07%±1.45%(P＜0.05).ELISA검측상응IL-4、IFN-γ적분비,표준변응원조IL-4위(74.61±13.82)pg/ml、(25.31±7.23)pg/ml;LPS조분별위(28.57±3.36)pg/ml、(163.41±82.37)Pg/ml.결론 AR환자중표체TIM-1적Th세포고우건강자,DC제정항원후가이촉진기표체;TIM-1적표체유리우초시Th세포향Th2분화,유차유발이Th2세포인자위주적면역반응.
Objective To investigate the significance of T cell immunoglobulin domain and mucin domain-1 (TIM-1) expression in helper T lymphocyte in allergic rhinitis (AR) patients.Methods Peripheral blood samples were collected from 20 AR patients, 9 males and 11 females, aged 31.4 (20 -54), and 20 healthy persons, 12 males and 8 females, aged 38,6 (18 -60).Peripheral blood mononuclear cells (PBMNs) were isolated and magnetic cell sorting technique was used to purify the CD4 + T cells and CD4 + CIM5RA + T cells, lmmunofluorescence staining and flow cytometry were used to detect the percentage of CD4 + CD45RA + T cells positive in TIM-1, a helper T lymphocyte marker.Phytohemagglutiniun (PHA) was added into the culture media to stimulate the T1M-1 positive helper T lymphocytes.Dendritic cells (DCs) thus induced and differentiated were conditioned by standardized allergen from dust mite (Dermatophagoides pteronyssinus) or lipopolysaccharide (LPS), and then co-cultured with CD4 T cells.The percentage of TIM-1 positive CD4 + CD45RA + T cells was counted.ELISA was used to detect the IL-4 and interferon (IFN) -γ levels in the supernatants of the culture media of the dust mite and LPS groups.Results The percentage of TIM-1 positive CD4 + T cells of the AR patients was (9.48 ± 1.51) %, significantly higher than that of the healthy controls[(3.12 ± 0.32) %, P < 0.05], and the percentage of TIM-1 positive CD4 + T cells after PHA stimulation of the AR patients was (47.05 ± 4.35) %, significantly higher than that of the healthy controls[(29.92 ± 3.36) %, P < 0.05].The percentage of TIM-1 positive CD4 + CD45RA + T cells of the AR patients was (26.34 ± 4.0)%, significantly higher than that of the healthy controls[(18.4 ± 8.1) %, P < 0.05], and the percentage of TIM-I positive CD4 + CD45 RA + T cells after PHA stimulation of the AR patients was (65.3 ± 7.4) %, significantly higher than that of the healthy controls[(36.3 ± 6.8) %, P < 0.05].The percentage of TIM-1 positive CD4 + CD45RA + T cells co-cultured with the DCs conditioned by dust mite of the AR patients was (10.81 ± 1.48) %, significantly higher than that of the LPS group[(6.08 ± 1.45) %]and the blank control group[(1.56 ± 2.30) %, both P < 0.05].The IL-4 level of the DCs conditioned by dust mite allergen was (74.61 ± 13.82) pg/ml, significantly higher than those of the LPS group and blank control group[(28.57 ± 3.36) and (1.40 ± 0.99) ng/ml respectively, both P < 0.05].The IFN-γ level of the DCs conditioned by dust mite allergen was (25.31±7.23) pg/ml, significantly lower than that of the LP3 group[(163.41 ± 82.37) pg/ml, P < 0.05], however, significantly higher than that of the blank control group[(1.58 ± 0.53) pg/ml, P < 0.05].Conclusion The percentage of TIM-1 positive helper T lymphocytes of the AR patients are abnormally high.After stimulation of allergen The DCs increase the number of TIM-1 expressing naive Th cells and promote them to differentiate into Th2 cells.Promoting naive Th cells to differentiate into Th2 cells, TIM-1 expression induces Th2 cytokine-based immune responses.
