乙肝疫苗负载的树突状细胞诱导慢性乙型肝炎患者特异性免疫应答

Proliferation of peripheral blood dendritic cell in patients with chronic hepatitis B and immunological response of injection of dendritic cells

目的 探讨自体乙肝疫苗致敏的树突状细胞(DC)回输慢性乙型肝炎(CHB)患者后的免疫应答效果。方法取患者外周血20ml分离单个核细胞,加入重组人粒细胞-巨噬细胞集落刺激因子(rGM-CSF)和白细胞介素4(IL-4)进行DC体外扩增,于培养第5天加入50μg／ml乙型肝炎疫苗,7天收获细胞。34例慢性乙肝患者根据年龄和感染病毒时间分为治疗1、2、3组,皮内回输DC;20例对照组CHB注射等量生理盐水,与治疗组均每周一次,连续8次。于回输DC后2周检测患者血清HBV-DNA含量及外周血CD3+、CD4+、CD8+T淋巴细胞。结果 3个治疗组患者回输DC后血清HBV-DNA含量拷贝数均较对照组及回输前显著降低(P<0.01),总应答率58.8％(20／34),对照组输注前后没有明显变化(P>0.05);治疗1组与3组相比HBV-DNA定量拷贝数降低幅度有显著性差异(P<0.01),治疗1组与2组、2组与3组相比均无显著性差异(P>0.05)。回输DC后患者CD4+细胞较回输前增高显著(P<0.01),CD3+、CD8+细胞增高不明显(P>0.05)。结论 经乙型肝炎疫苗致敏的DC回输CHB患者后可产生明显应答,且应答程度与患者感染病毒时间和／或年龄有一定关系。

Objective To investigate the immunological effect of DCs on patients with chronic hepatitis B. Methods Peripheral blood of patients with chronic hepatitis B.DCs prodiferated from PBMCs by adding rhGM - CSF and IL-4 were cultured for seven davs.On the fifth day, DCs were added to 50μg/ml hepatitis B vaccine. 34 patients with hepatitis B injected by DCs, regarded as 3 observation groups, for eight times wddkly, while the 20 patients in control group given equal volum NS. HBV - DNA contents in the serum and T cell of CE3+ , CD4 + , CD8 + in the peripheral blood of patients were tested after the injection course. Result After treatment of DCs, the copies of HBV - DNA in the serum decreased greatly(P<0.01) and the total effective rate is 58.8% (20/34) .No significant changes were found after injection of DCs in the control group( P > 0.05) . There was great difference in the copies of HBV - DNA between the first observation group and the third observation group( P < 0.01). The others were not significant different (P > 0.05). After injection of DCs, T cells of CD4+ decreased greatly(P < 0.01), while T cells of CD3+ , CD8 + were not significant different( P > 0.05) . Conclusion The patients with HBV have significant response by the injection of DCs. And degree of response may be related to the time of infection virus and/or the age of patient.