HBe Ag对髓样树突状细胞Toll样受体4及NF-κB信号通路的影响

Effect of HBeAg on toll like receptor 4 and NF-κB signaling pathway in human dendritic cells

目的探讨HBe Ag对髓样树突状细胞(mDC)Toll样受体4(TLR4)及NF-κB信号通路的影响。方法将健康者外周血单核细胞诱导分化为未成熟mDC;加入HBe Ag体外刺激,经脂多糖(LPS)刺激获得成熟mDC。采用Western blot法检测TLR4、NF-κB信号通路蛋白相对表达量,MTS法检测mDC刺激同种异体淋巴细胞增殖能力。结果培养至第9天,HBe Ag刺激组TLR4、NF-κB相对表达量为0.12±0.01、0.75±0.12,分别低于对照组的0.27±0.03、1.20±0.13(均P<0.05)。在DC/淋巴细胞比例为1∶5、1∶10、1∶20的反应中,HBe Ag刺激组刺激指数为2.93±0.05、2.56±0.19、1.44±0.09,分别低于对照组的4.83±0.05、3.57±0.35、2.13±0.11(均P<0.05)。结论 HBe Ag抑制mDC TLR4信号通路蛋白的表达,减弱mDC刺激同种异体淋巴细胞增殖能力。

Objective To investigate the effect of HBeAg on the toll-like receptor 4(TLR4) and nuclear factor κB(NF-κB)signaling pathway in human monocyte-derived dendritic cells(m DCs). Methods The peripheral blood mononuclear cells were isolated from healthy volunteers, and then induced and proliferated to immature m DC in vitro. Mature m DCs were obtained after HBeAg and lipopolysaccharide stimulation. The expression of TLR4 and NF-κB proteins was detected wit Western blot, and the response of allogeneic lymphocytes was examined with MTS method. Results The expression of TLR4 and NF-κB proteins decreased in the HBeAg-stimulated groups( 0. 12 ± 0. 01, 0. 75 ± 0. 12) compared with that of control group(0.27 ±0.03, 1.20±0.13, P<0.05). In response to the DC/lymphocyte with ratio of 1:5, 1:10 and 1:20, the stimulation indexes of the HBeAg stimulation group was 2.93 ±0.05,2.56 ±0.19, 1.44 ±0.09 respectively, which were lower than that of the control group(4.83 ±0.05, 3.57±0.35, 2.13 ±0.11, P<0.05).Conclusion HBeAg can inhibit TLR4 and NF-κB signaling pathway of m DC, weaken the ability of stimulating T cell proliferation.