丙型肝炎病毒F蛋白体外免疫调节作用  

Immunomodulatory effects in vitro of F protein of hepatitis C virus

作　　者：郭亚光[1] 丁丰[1] 邵凌云[2] 沈华江[1] 

机构地区：[1]浙江省绍兴市第六人民医院肝病科312000 [2]复旦大学附属华山医院感染科,上海200040

出　　处：《国际流行病学传染病学杂志》2015年第4期232-236,共5页International Journal of Epidemiology and Infectious Disease

摘　　要：目的探讨HCVF蛋白对DC及T细胞的免疫调节作用。方法构建HCVF蛋白的表达菌株，并表达、纯化F蛋白；收集42例慢性丙型肝炎患者及30例健康对照，检测血清抗F蛋白抗体的阳性率；分离PBMC，并体外诱导分化DC细胞，用纯化的F蛋白刺激DC细胞，流式细胞术检测DC细胞表面CD69、CD80、CD95、CD95L的表达情况；用纯化的F蛋白刺激PBMC，ELISA方法检测细胞上清中IFNγ、IL-12、IL-4、IL-10的含量。结果成功克隆表达F蛋白，纯度较高（浓度0．8mg／mL，纯度大于90％）；42例慢性丙型肝炎患者m清中特异性抗F蛋白抗体阳性率为42．88％（18／42）；F蛋白可诱导DC细胞表面CD69、CD80、CD95L、CD95明显上调．且丙型肝炎患者这些分子的表达水平明最高于健康对照组（t=2．824、3．707、4．788、3．550，P均〈0．05）。F蛋白诱导丙型肝炎患者PBMC产生的Th1型细胞因子IFNγ、IL-12含量明显低于健康对照组（t=4．583、3．708，P均〈O．05），而Th2型细胞因子IL-4、IL-10含量明显高于健康对照组（t=3．483、2．872，P均〈0．05）。结论丙型肝炎F蛋白可诱导机体产生特异性抗体，上凋DC细胞表面活化及凋亡分子．并可促进Th1／Th2失衡。Objective To investigate the immunomodulatory effects ofF protein of hepatitis C virus to dentritic cells （DC） and T cells. Methods F gene of hepatitis C virus was cloned and F protein was expressed and purified. A total of 42 patients with chronic hepatitis C （CHC） and 30 healthy controls were enrolled in this study to detect the positive rate of serum anti-F protein. We separated the peripheral blood mononuclear cells （PBMC）, and induced to differentiate DC cells in vitro. DC was stimulated by F protein and the expression levels of CD69, CD80, CD95, CD95L were detected by flow cytometry. The PBMC were stimulated by purified F protein and the cytokines like IFNγ IL-12, IL-4, IL-10 in the culture supernatant were detected by ELISA. Results High purity （0.8 mg/mL, purity〉90% ） of F protein was produced in this study. The prevalence of anti-F protein in these 42 CHC patients was 42.88% （18/42）. CD69, CDS0, CD95, CD95L on DC was significantly up-regulated after stimulation by F protein, and the expression level of these markers were all higher in CHC patients comparing with healthy controls （t=2.824, 3.707, 4.788, 3.550 respectively, all P〈0.05）. Thl cytokines like IFNγ and IL-12 produced by PBMC in CHC patients after stimulation were much lower than healthy controls （t=4.583, 3.708 respectively, all P〈0.05）, but Th2 cytokines IL-4 and IL-10 were much higher than healthy controls （t=3.483, 2.872 respectively, all P〈0.05）. Conclusions HCV F protein is able to induce production of specific anti-F antibodies, up-regulate the expression of makers of activation and apoptosis, and promote the imbalance of Th1/Th2 cytokines.

关 键 词：肝炎病毒 F蛋白 DC细胞 T细胞 

分 类 号：R373.21[医药卫生—病原生物学]