治疗性乙型肝炎疫苗的免疫效应

Immune response for phase I clinical trial of a hepatitis B immunogenic complex therapeutic vaccine, YIC

目的观察治疗性乙型肝炎（乙肝）疫苗（重组乙肝表面抗原和高效价抗乙肝免疫球蛋白组成的免疫复合物，简称乙肝疫苗）Ⅰ期临床研究中诱生的免疫应答作用。方法用间接酶联免疫吸附试验（ELISA）测定乙肝疫苗诱生的抗乙型肝炎表面抗原（HBSAg）抗体免疫球蛋白亚类；用。HTdR掺入法进行非特异性和特异性淋巴细胞转化试验；用流式细胞术检测细胞因子，使用BD-CBA试剂盒。结果所有志愿者中，乙肝疫苗诱导产生抗HBSAg抗体，其IgG免疫球蛋白亚类结果显示，所有受试者（20／20）为IgG1、IgG3双阳性和IgG2和IgG4双阴性，IgG1、IgG3双阳性者中IgG1均强于IgG3，但每个受试者之间强弱程度有所不同。淋巴细胞转化试验结果显示，80％（4／5）的志愿者重组HBSAg特异性淋转刺激指数（SI）大于2．0，植物血凝素非特异性淋转刺激指数和重组HBSAg特异性淋转刺激指数无相关性。细胞因子检测结果显示，第一次注射乙肝疫苗后42d，干扰素v，白细胞介素（IL）-2、IL-4、IL-6、IL-10和肿瘤坏死因子α和注射前相比，没有显著变化，而在71d时所有志愿者（8／8）干扰素γ和7名志愿者（7／8）IL-2高于注射前（P值〈0．01或P〈0．05）；而其他4种细胞因子在注射后71d没有显著性变化（P值均〉0．05）。结论治疗性乙型肝炎疫苗在健康受试者中有效地诱导出特异性体液免疫和细胞免疫。

Objective A hepatitis B immunogenic complex therapeutic vaccine, yeast-derived recombinant HBsAg combined with human anti-HBs immunoglobulin （YIC）, was evaluated for safety and immune response in phase Ⅰ clinical trial. Methods The subtypes IgG1, IgG2, IgG3 and IgG4 of serum anti-HBs collected from 20 immunized subjects were analyzed by ELISA. The lymphocyte proliferation assay was carried out in five subjects and was analyzed by ^3Hthymidine incorporation. The assays for IFN7, IL-2, IL-4,IL-6, IL-10 and TNFα were measured using Human Cytometric Bead Array Kit with FACSCalibur. Results The results showed that the subtypes of anti-HBs antibodies induced by 30, 60 and 90 p.g YIC-immunized groups among all of the adult volunteers （20/20） were IgG1 and IgG3. The level of IgGl was higher than that of IgG3 in each volunteer but the strength was different from each other. The rHBsAg-stimulated lymphocyte proliferation induced by three injections of 90 I.tg of YIC showed that the stimulation index was more than 2.0 in four out of the five individuals （4/5）, ranging from 2.70 to 4.75. PHA-stimulated lymphocyte proliferation was not related to rHBsAg-stimulated lymphocyte proliferation. In the 60μg YIC-immunized group there was no significant difference between the levels of IFN7, IL-2, IL-4, IL-6 and IL-10 at day 0 and day 42. At day 71, in comparison to day 0, the level of IFN7 was higher in all eight subjects studied （ P = 0.015 ） and the level of IL-2 was also increased in seven out of eight subjects （ P = 0.002 ）. In contrast, the levels of IL-4, IL-6, IL-10 and TNFα showed no significant difference in all the subjects （ P-values： 0.298, 0.976, 0.202 and 0.996）. Conclusion Our results indicate that this hepatitis B immunogenic complex therapeutic vaccine （YIC） can induce a potent anti-HBs response.