Objective To observe the effect of interferon (IFN) λ1 on the function of peripheral blood mononuclear cell (PBMC) derived dendritic cells (DC) in adolescents with chronic hepatitis B virus (HBV) infection.Methods A total of 34 adolescent patients with chronic HBV infection including 16 cases in immune clearance phase and 18 cases in immune tolerant phase,and 10 healthy youths were enrolled in the study.PBMC from all samples were isolated and then cultured according to the following 3 groups.IFN-λ1 group:cultured with IFN-λ1 only; normal group:cultured with recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) and recombinant human interleukin-4 (rhIL-4); combination group:cultured with IFN-λ1,rhGM-CSF and rhIL-4.The shape of DC was observed and the phenotypic patterns of CD83,CD80,CD86 and human leucocyte antigen (HLA)-DR were characterized by flow cytometry.The levels of interleukin 12 (IL-12) and IFN-γ in the supernatant produced by DC were analyzed with enzyme-linked immunosorbent assay (ELISA).Independent-samples t test was used to compare means between two groups and one-way ANOVA was used to compare means among multiple groups.Results In the IFN-λ1 group,the shape of adherent cells showed no change after 7 days of culture.In the normal group and combination group,typical morphology of mature state DC was observed.The expressions of phenotypic patterns of CD83,CD80,CD86 and HLA-DR in DC from immune tolerant phase patients were lower than those from healthy control youths and immune clearance phase patients (F=14.82-46.32,all P＜0.01).Co-stimuation of IFN-λ1 with rhGM-CSF and rhIL-4 up-regulated the expressions of phenotypic patterns on DC.Among patients both in immune tolerant phase and immune clearance phase,the expressions of CD83,CD80,CD86 and HLA-DR in the combination group were higher than those in normal group (t=3.16-5.69,all P＜0.01).In the normal group,the levels of IL-12 and IFN-γ in culture supernatants from healthy aldolescents were (287.25 ± 19.78) pg/mL and (56.38 ± 15.27)pg/mL,respectively; those from patients in immune tolerant phase were (198.42±22.28) pg/mL and (29.36±8.45) pg/mL,respectively; those from patients in immune clearance phase were (256.12±18.43) pg/mL and (38.38±9.72) pg/mL,respectively (F=69.22 for IL-12,F=20.29 for IFN-γ,both P＜0.01).IFN-λ1 combined with rhGM-CSF and rhIL-4 significantly up-regulated the secretion of IL-12 and IFN-γ by matured DC from patients both in immune tolerant phase and immune clearance phase (t=4.69-8.55,all P＜0.01).Conclusions In adolescent patients with chronic HBV infection,the DC function is markedly impaired in those in immune tolerant phase,compared with patients in immune clearance phase and healthy youths.IFN-λ1 can promote the maturation and phenotypic pattern expression of PBMC derived DC in adolescents with chronic HBV infection both in immune tolerant phase and immune clearance phase.%目的 探讨干扰素λ1对慢性HBV感染青少年患者外周血单个核细胞(PBMC)来源树突状细胞(DC)功能的影响.方法 选取慢性HBV感染青少年患者34例,其中免疫耐受期18例,免疫清除期16例；设健康对照者10名.采集外周血分离PBMC,分3组体外培养PBMC诱导成熟DC,干扰素λ1培养组:仅加入干扰素λ1；常规培养组:加入重组人IL-4和重组人粒细胞巨噬细胞集落刺激因子(GM-CSF)；联合培养组:加入IFN-λ1、IL-4和GM-CSF共同培养.7d后观察各组DC形态,用流式细胞仪检测CD80、CD83、CD86和人白细胞抗原DR(HLA-DR)表达情况,ELISA法检测上清液INF-γ和IL-12水平.多组间比较应用one way ANOVA,两组比较采用t检验.结果 DC培养7d后,干扰素λ1培养组未见贴壁细胞有形态改变,常规培养组和联合培养组可见典型的成熟DC形态.常规培养组和联合培养组免疫耐受期患者DC表面分子CD80、CD83、CD86和HLA-DR表达量均低于健康对照者和免疫清除期患者(F值14.82～46.32,均P＜0.01)；经干扰素λ1联合诱导能促进免疫耐受期和免疫清除期患者DC高表达上述表面分子,与常规培养组比较,差异有统计学意义(t值3.16～5.69,均P＜0.01)；健康对照者、免疫耐受期和免疫清除期患者常规培养组DC培养上清液IL-12水平分别为(287.25±19.78)、(198.42±22.28)和(256.12±18.43)pg/mL; IFN-γ水平分别为(56.38±15.27)、(29.36±8.45)和(38.38±9.72) pg/mL(F值分别为69.22和20.29,均P＜0.01).干扰素λ1联合诱导能促进免疫耐受期和免疫清除期患者DC培养上清液IL-12和IFN-γ水平(t值4.69～8.55,均P＜0.01).结论 HBV慢性感染青少年免疫耐受期患者DC功能较健康人和免疫清除期患者明显低下,IFN-λ1可促进免疫耐受期和免疫清除期患者PBMC来源DC成熟和共刺激分子表达.
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