抗HBV-DC治疗e抗原阳性慢性HBV感染者的初步临床研究

Clinical Research on Anti-HBV Dendritic Cell on the HBeAg Positive Patients with Chronic HBV Infection

目的观察HBsAg致敏患者自体外周血单个核细胞来源的树突状细胞治疗e抗原阳性慢性乙型肝炎病毒感染者的近期临床效果。方法对9例轻度慢性乙型肝炎患者及15例慢性HBV携带者进行临床试验。取外周静脉血25mL,用密度梯度离心及贴壁法获得单核细胞,以粒细胞-巨噬细胞集落刺激因子和白细胞介素4诱导扩增出DC,并予HBsAg致敏DC。第7天收获DC静脉回输,每周1次,平均4次/例。治疗前后检测HBV标志物定量、HBV-DNA定量及肝功能,两次检测平均间隔37d。结果好转23例,无效1例,有效率为95.83%。治疗前后HBsAg为(235.37±87.26)ng/mL和(90.10±82.30)ng/mL(t=7.66,P<0.001),平均下降62%;HBeAg为(2.74±1.74)Ncu/mL和(1.65±1.37)Ncu/mL(t=4.08,P<0.001),平均下降48%;HBeAb为(0.23±0.45)Ncu/mL和(0.75±0.97)Ncu/m(Lt=3.22,P<0.01);3例HBeAb由阴转阳,1例发生HBeAg/HBeAb血清学转换;HBV-DNA定量为(1.30±1.45)×108copy/mL和(0.53±1.27)×108copy/mL(t=2.20,P<0.05),平均下降1.04log。结论抗HBV-DC静脉回输可有效抑制e抗原阳性慢性HBV感染者的病毒复制,减少病毒载量,降低或清除病毒抗原,促进HBeAb的产生并促进HBeAg/HBeAb的血清学转换。慢性HBV携带者和CHB患者均可产生应答反应,是一种安全、有效的治疗e抗原阳性慢性HBV感染者的方法,显示出良好的应用前景。

Objective To observe the clinical effect of autalogous HBsAg-loaded dendritic cells on the HBeAg positive patients with chronic HBV infection. Methods Monocytes were isolated from peripheral blood of 9 chronic hepatitis B and 15 chronic HBV carriers by Ficoll-Hypaque density gradient centrifugation and cultured with plastic-adherence method. DCs were induced and proliferated from the monocytes with granulocyte-macrophage clony stimulating factor and interleukin-4 for 7 days. After being incubated with HBsAg, DCs were injected to patients＇ vein for average 4 times at one week＇s interval. HBV markers, HBV DNA level, and liver function tests in the serum of patients were tested before and after treatment. Results 23 of the 24 （95.83%） patients responded clinically. The HBsAg leveis were （235.37 ± 87.26） ng/mL and （90.10 ± 82.30） ng/mL （t = 7.66,P 〈0.001）, the HBeAg levels were （2.74 ± 1.74） Ncu/mL and （1.65 ± 1.37） Ncu/mL（t = 4.08,P 〈0.001）, the HBeAb levels were （0.23 ± 0.45） Ncu/mL and （0.75± 0.97） Ncu/mL（t = 3.22, P 〈0.01）, and the HBV DNA levels were （1.30 ± 1.45） ×10^8 copy/mL and （0.53± 1.27） ×10^8 copy/mL（ t = 2.20, P 〈0.05 ） before and after treatment, respectively. The HBeAg of 3 patients was cleared, and the HBeAg/HBeAb seroconversion occurred in one patient. Conclusion The autologous anti-HBV DCs can effectively suppress HBV replication, reduce virus and virus antigen load in serum, eliminate HBeAg, increase HbeAb level and promote HBeAg/HBeAb seroconversion. CHB patients and chronic HBV carrier also are more effective and more safe treatmen for anti-HBV DCs.