目的分析慢性乙型肝炎患者外周血树突状细胞(DC)上Toll样受体3(TLR3)的表达变化及其临床意义。方法随机筛选慢性乙型肝炎(CHB)患者20例,并以10例健康者作对照,分别从外周血分离单个核细胞,体外诱导培养DC。流式细胞仪(FCM)测定未成熟期DC(imDC)与成熟期DC(mDC)的表型;分别应用流式细胞仪和Western印迹法测定imDC与mDC上TLR3的表达率和受体蛋白的表达。结果在健康对照组,mDC表面分子CD80、CD86、组织相容性白细胞抗原(HLA)-DR、CD83的表达率分别为(82、35±8.67)%、(79.61±10.08)%、(92.79±8、48)%和(83.76±5.47)%,imDC表面分子表达率分别为(28、31±8.79)%、(31.17±11.23)%、(27.61±10.28)%和(23.46±11.53)%,各组间比较,差异均有统计学意义(P〈0.01);而在CHB组,mDC与imDC表面分子表达率分别为(42.46±9.22)%、(40.72±11.24)%、(48.57±12.51)%、(22.25±11.22)%和(18.57±10.22)%、(24.16±10.46)%、(17.87±10.38)%、(14.28±6.77)%,各组间比较,差异均无统计学意义(P〉0.05);健康者组mDC上表面分子的表达率与CHB患者组比较,各组间差异均有统计学意义,而两组受试者imDC上表面分子的表达率比较,差异均无统计学意义。健康对照imDC上TLR3的表达率明显高于mDC(P〈0.01),而CHB患者imDC与mDC上TLR3的表达率差异无统计学意义(P〉0.05);且CHB患者imDC上TLR3的表达率明显低于健康对照(P〈0.01)。结论CHB患者外周血DC上存在TLR3的表达下调,可能是HBV感染慢性化的重要原因。
Objective To investigate the expression of Toll-like receptor 3 (TLR3) on dendritic cells(DCs) derived from peripheral blood mononuclear cells(PBMCs) of chronic hepatitis B(CHB) patients and to explore the mechanism of sustained infection of hepatitis B virus (HBV). Methods Twenty CHB patients were randomly screened in the study, and ten healthy persons were recruited as controls. The monocytes isolated from peripheral blood of candidates were incubated with recombinant human granulocyte macrophage colony-stimulating factor (rhGM-CSF) and rhIL-4 to induce the DCs generation and proliferation. Then the phenotype of DCs was identified by microscope. The expressions of the phenotypes[histocompatihility leukocyte antigen(HLA)-DR, CD80, CD86, CD83] of immature and mature DCs were measured by flow cytometer. Furthermore, the expression of TLR3 on mature DCs(mDC) and immature DCs(imDC) was determined by flow cytometry and Western blot analysis respectively. Results As for healthy volunteers, the expressions of CD80, CD86, HLA-DR and CD83 on DCs at the 7th day, which were(82.35 ± 8. 67)%, (79.61 ±10.08)%, (92.79±8.48) % and (83.76± 5.47) % respectively, were significantly higher than those at the 5th day which were(28. 31 ± 8.79) %, (31.17± 11.23) %, (27.61 ±10.28) % and (23.46 ± 11.53) % respec tively(P 〈 0. 01). However, no difference was found between mDC and imDC of the CHB patients groups (P 〉 0.05). The expression of TLR3 on imDC was significantly higher than that on mDC at control group (P 〈 0.01), but no significant difference was shown in CHB patients(P 〉 0.05). And the expression of TLR3 on imDC in CHB patients group was significantly lower than that of control group(P 〈 0.01). Concision The expression of TLR3 on peripheral DCs of CHB patients is decreased compared to that of healthy persons, it might be one of the important pathogenesis mechanisms of HBV st,stained infection.