中华传染病杂志
中華傳染病雜誌
중화전염병잡지
CHINESE JOURNAL OF INFECTIOUS DISEASES
2014年
2期
94-99
,共6页
燕飞%汤永志%潘柯传%朱坚胜%朱敏%陈华忠%赵海红%刘均艳%陈景丹
燕飛%湯永誌%潘柯傳%硃堅勝%硃敏%陳華忠%趙海紅%劉均豔%陳景丹
연비%탕영지%반가전%주견성%주민%진화충%조해홍%류균염%진경단
树突状细胞%肝炎表面抗原,乙型%Toll样受体4%核因子-κB%丝裂原激活蛋白激酶类%信号传导
樹突狀細胞%肝炎錶麵抗原,乙型%Toll樣受體4%覈因子-κB%絲裂原激活蛋白激酶類%信號傳導
수돌상세포%간염표면항원,을형%Toll양수체4%핵인자-κB%사렬원격활단백격매류%신호전도
Dendritic cells%Hepatitis B surface antigen%Toll-like receptor 4%NF-κB%Mitogenactivated protein kinase%Signal transduction
目的 探讨HBsAg对脂多糖刺激成熟的单核细胞来源的树突状细胞(MoDC)功能和细胞表面TLR4信号通路的影响.方法 将健康者PBMC诱导分化为未成熟MoDC,加入不同浓度HBsAg体外刺激,再经脂多糖刺激获得成熟MoDC.Western印迹检测各组TLR4、核因子-κB、丝裂原激活蛋白激酶信号通路蛋白表达,ELISA检测细胞因子,细胞增殖与毒性检测试剂盒检测刺激同种异体淋巴细胞增殖的能力.两组间比较采用t检验,多组间比较采用单因素方差分析.结果 未经HBsAg刺激的对照组TLR4、核因子-κB、pp38、磷酸化c-Jun氨基末端激酶1/2 (pJNK1/2)蛋白分别为0.43±0.14、0.69±0.27、0.21±0.09和0.44±0.13,5μg/mL HBsAg刺激后分别增至1.22±0.39、1.37±0.40、0.74±0.17和0.92±0.15(t值分别为-3.14、-3.09、-5.07和-2.93,均P＜0.05);但细胞外信号调节激酶1/2表达下降(2.43±0.80比1.13±0.23,t=3.18,P＜0.05).5μg/mL HBsAg刺激后的MoDC刺激同种异体T淋巴细胞,其刺激指数高于对照组,且随MoDC/淋巴细胞比值增高而增高(t值分别为-5.72、-4.93和-4.96,均P＜0.05).5μg/mL HBsAg刺激后MoDC分泌IL-12、TNF-α分别为(637.8±19.2)和(710.6±47.4)pg/mL,对照组分别为(597.8±15.4)和(568.5±35.0)pg/mL(t值分别为-5.07和-6.70,均P＜0.05).结论 HBsAg可激活MoDC表面TLR4信号通路,促进细胞因子分泌,增强刺激T淋巴细胞增殖的能力.
目的 探討HBsAg對脂多糖刺激成熟的單覈細胞來源的樹突狀細胞(MoDC)功能和細胞錶麵TLR4信號通路的影響.方法 將健康者PBMC誘導分化為未成熟MoDC,加入不同濃度HBsAg體外刺激,再經脂多糖刺激穫得成熟MoDC.Western印跡檢測各組TLR4、覈因子-κB、絲裂原激活蛋白激酶信號通路蛋白錶達,ELISA檢測細胞因子,細胞增殖與毒性檢測試劑盒檢測刺激同種異體淋巴細胞增殖的能力.兩組間比較採用t檢驗,多組間比較採用單因素方差分析.結果 未經HBsAg刺激的對照組TLR4、覈因子-κB、pp38、燐痠化c-Jun氨基末耑激酶1/2 (pJNK1/2)蛋白分彆為0.43±0.14、0.69±0.27、0.21±0.09和0.44±0.13,5μg/mL HBsAg刺激後分彆增至1.22±0.39、1.37±0.40、0.74±0.17和0.92±0.15(t值分彆為-3.14、-3.09、-5.07和-2.93,均P＜0.05);但細胞外信號調節激酶1/2錶達下降(2.43±0.80比1.13±0.23,t=3.18,P＜0.05).5μg/mL HBsAg刺激後的MoDC刺激同種異體T淋巴細胞,其刺激指數高于對照組,且隨MoDC/淋巴細胞比值增高而增高(t值分彆為-5.72、-4.93和-4.96,均P＜0.05).5μg/mL HBsAg刺激後MoDC分泌IL-12、TNF-α分彆為(637.8±19.2)和(710.6±47.4)pg/mL,對照組分彆為(597.8±15.4)和(568.5±35.0)pg/mL(t值分彆為-5.07和-6.70,均P＜0.05).結論 HBsAg可激活MoDC錶麵TLR4信號通路,促進細胞因子分泌,增彊刺激T淋巴細胞增殖的能力.
목적 탐토HBsAg대지다당자격성숙적단핵세포래원적수돌상세포(MoDC)공능화세포표면TLR4신호통로적영향.방법 장건강자PBMC유도분화위미성숙MoDC,가입불동농도HBsAg체외자격,재경지다당자격획득성숙MoDC.Western인적검측각조TLR4、핵인자-κB、사렬원격활단백격매신호통로단백표체,ELISA검측세포인자,세포증식여독성검측시제합검측자격동충이체림파세포증식적능력.량조간비교채용t검험,다조간비교채용단인소방차분석.결과 미경HBsAg자격적대조조TLR4、핵인자-κB、pp38、린산화c-Jun안기말단격매1/2 (pJNK1/2)단백분별위0.43±0.14、0.69±0.27、0.21±0.09화0.44±0.13,5μg/mL HBsAg자격후분별증지1.22±0.39、1.37±0.40、0.74±0.17화0.92±0.15(t치분별위-3.14、-3.09、-5.07화-2.93,균P＜0.05);단세포외신호조절격매1/2표체하강(2.43±0.80비1.13±0.23,t=3.18,P＜0.05).5μg/mL HBsAg자격후적MoDC자격동충이체T림파세포,기자격지수고우대조조,차수MoDC/림파세포비치증고이증고(t치분별위-5.72、-4.93화-4.96,균P＜0.05).5μg/mL HBsAg자격후MoDC분비IL-12、TNF-α분별위(637.8±19.2)화(710.6±47.4)pg/mL,대조조분별위(597.8±15.4)화(568.5±35.0)pg/mL(t치분별위-5.07화-6.70,균P＜0.05).결론 HBsAg가격활MoDC표면TLR4신호통로,촉진세포인자분비,증강자격T림파세포증식적능력.
Objective To investigate the effects of hepatitis B surface antigen (HBsAg) on the tolllike receptor 4 (TLR4) signaling pathway of lipopolysaccharide (LPS)-stimulated human monocytederived dendritic cells (MoDC).Methods The peripheral blood mononuclear cells (PBMC) were isolated from healthy volunteers,and then induced and proliferated to immature MoDC in vitro.Mature MoDC were obtained by HBsAg and LPS stimulation.The expressions of TLR4,nuclear factor κB (NF-κB) and mitogen-activated protein kinase (MAPK) protein were determined by Western blotting assay,and the concentrations of cytokines in the supermatant were detected by enzyme-linked immunosorbent assay (ELISA).In addition,the capacity of MoDC to stimulate allogenic lymphocyte was measured by MTS assay.Statistical analyses were performed by t test and analysis of variance.Results The expressions of TLR4,NF-κB,phosphorylation p38 (pp38),phosphorylation c-Jun N-terminal kinase 1/2 (pJNK1/2) protein were upregulated in the HBsAg-stimulated groups with 5 μg/mL HBsAg compared with those of control group (0.43±0.14 vs 1.22±0.39,0.69±0.27 vs 1.37±0.40,0.21±0.09 vs 0.74±0.17,and 0.44±0.13 vs 0.92±0.15,t=-3.14,-3.09,-5.07,-2.93,respectively,all P＜0.05),while the expression of phosphorylation extracellular signal-regulated kinase 1/2 (pERK 1/2) was downregulated in HBsAg stimulated groups,which were compared with that of control group (2.43±0.80 vs 1.13±0.23,t=3.18,P＜0.05).The capacity of MoDC treated with 5 μg/mL HBsAg to stimulate allogenic T lymphocytes was enhanced,and the stimulation index increased with DC-to-lymphocyte ratio (t=-5.72,-4.93,-4.96,P＜0.05).In addition,the 5 μg/mL HBsAg-pulsed MoDC were found to have a stronger capacity to produce interleukin (IL)-12 and tumor necrosis factor-α (TNF-α) compared with those of control group ([637.8±19.2] pg/mL vs [597.8±15.4] pg/mL and [710.6±47.4] pg/mL vs [568.5± 35.0] pg/mL,respectively; t=-5.07,-6.70,both P＜0.05).Conclusion HBsAg can activate TLR4 signaling pathway of MoDC,promote the secretion of cytokines and enhance the ability of MoDC stimulating T cell proliferation.
