干扰素抗病毒应答与慢性乙型肝炎患者外周血树突状细胞功能的关系

目的:探讨干扰素抗病毒应答与慢性乙型肝炎患者外周血树突状细胞(DCs)功能的关系。方法:分别采集23例慢性乙型肝炎患者干扰素治疗前和治疗满4mo时的抗凝外周静脉血,分离外周血单个核细胞(PBMCs),在重组人白细胞介素4和重组人粒细胞-巨噬细胞集落刺激因子的作用下培养7 d使DCs增殖、成熟。以间接免疫荧光流式细胞技术检测DCs的表型;以ELISA法检测DCs单独培养上清液中IL-12的水平;用DCs与HBsAg共同孵育,丝裂霉素C处理后,再与自体PBMCs共同培养,加入H-TDR,收集细胞测定cpm值。实验中以8例正常健康人作为对照。结果:干扰素完全应答组DCs表面CD,HLA-DR和ICAM-1的表达较治疗前均显著增加(均为P<0.05),且CD,CD,HLA-DR和ICAM-1的增高与无应答组相比具有显著性意义(分别为P<0.05、P<0.01、P<0.01和P<0.05);完全应答组DCs分泌IL-12的水平在治疗后显著增加(P<0.01);DCs的抗原提呈作用在治疗前、后,完全应答组比无应答组均显著增强(P<0.01和P<0.001),而部分应答与无应答组之间无显著性差异(P>0.05)。结论:慢性乙肝患者干扰素的抗病毒应答与外周血DCs的功能状态有关,干扰素能显著促进患者DCs功能的改善,进而可能增加患者对治疗的应答。

Study on the relationship between therapy response to interferon and the functional status of peripheral blood dendritic cells from patients with chronic hepatitis B

Objective: To study the relationship between therapy response to interferon and the functional status of peripheral blood dendritic cells (DCs) from patients with chronic hepatitis B. Methods: The peripheral blood samples were obtained from 23 patients at the time of interferon pretreatment and posttreatment for 4 months, respectively. The peripheral blood mononuclear cells (PBMCs) were isolated and cultured, and recombinant human IL-4 and GM-CSF were added to the cuLtures. After cuLtured for 7 days, DCs were harvested for the following studies. CD CD , HLA-DR as weLL as ICAM-1 on the membrane of DCs were detected by indirect immunofiuorescent flow cytometry. DCs were cultured for another 3 days, and the supernatants were collected and the levels of IL-12 were detected with ELISA. DCs were incubated with HBsAg, following treatment with mitomycin C, then DCs were collected and mixed with autogenous PBMCs and cocultured. At last, H TDR was added to the culture, and the cpm value was detected for all the harvested cells. 8 healthy individuals were regarded as controls. Results: in interferon complete responder group, the levels of CD , HLA-DR and ICAM-1 on DCs significantly increased in posttreatment group compared with those in pretreatment one (P<0.05 in all). Likewise, the levels of CD , CD , HLA-DR and ICAM-1 on DCs in complete responder group were significantly higher than those in nonresponder one, respectively (P0.05, P<0.01, P<0.01 and P<0.05). In complete responder group, the level of IL-12 secreted by DCs after treatment markedly increased than that before treatment (P<0.01). Regarding to the capacities of DCs antigen presenting in complete responder group, both in pretreatment group and in posttreatment one, they were significantly stronger than those in nonresponder group, respectively P<0.01 and P<0.001 . There was no significant difference between the partial responder group and the nonresponder one P>0.05 . Conclusions: The resuLts indicate that there may be a close relationship in the response to interferon therapy with the functional status of DCs from patients with chronic hepatitis B. Interferon therapy may markedly improve the functional status of DCs, and lead to facilitate the response to the antiviral treatment.