目的 研究慢性HBV感染时,DC-SIGN在树突状细胞(DC)成熟和活化中介导的作用. 方法 将α-甘露糖苷酶抑制剂-基夫碱作用于HepG2.2.15细胞,收获上清中的高甘露糖型HBV颗粒,并于第5 d加入到外周血单核细胞衍生的DC培养基中,培养至第7d,采用流式细胞仪检测DC表面CD1a、CD83、CD80、CD86、HLA-DR分子的表达,MTT法检测DC刺激淋巴细胞增殖的能力,ELISA法检测DC分泌IL-12的水平. 结果 高甘露糖型HBV组,与天然的HBV组相比,DC表面CD1a、CD83、CD80、CD86、HLA-DR分子的表达增加,分泌IL-12的水平升高,刺激同种异体淋巴细胞增殖的能力亦明显增强,且上述效应均可被DC-SIGN特异性抗体所阻断. 结论 DC-SIGN识别高甘露糖型HBV后可以促进DC的成熟和活化,天然的HBV可能利用α-甘露糖苷酶参与的去甘露聚糖修饰来逃避DC-SIGN的识别,从而诱导DC功能的缺陷.%Objective To investigate the role of DC-SIGN on the maturation and activation of dendritic cells (DC) in chronic HBV infection.Methods Highly mannosylated HBV obtained by treating HepG2.2.15 cells with the a-mannosidase I inhibitor kifunensine were added to the medium of DC derived from peripheral blood mononuclear cells on day 5 after culture.The expression rates of CDla, CD83, CD80, CD86 and HLA-DR of DCs were assayed by flow cytometry analysis, the stimulating capacity of DC was detected by MTT assay and the levels of IL-12 released by DCs were measured by ELISA on day 7.Results Compared with native HBV group, the expression of CDla, CD83, CD80, CD86 and HLA-DR of DC were significantly improved, levels of IL-12were increased, and capacity of stimulating lymphocyte proliferation was enhanced in highly mannosylated HBV group.Furthermore these effects could be all blocked by DC-SIGN specific antibody.Conclusion DC-SIGN can promote the maturation and activation of DC after recognizing highly mannosylated HBV, native HBV may exploit demannosylation as a way to escape recognition by DCSIGN and thereby induce DC dysfunction.
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