乙型肝炎病毒囊膜中蛋白与白细胞介素18联合基因免疫  

Preliminary study on hepatitis B virus nuclei acid vaccine with interleukin-18 as co-stimulator

作　　者：董菁[1] 成军[1] 王勤环[2] 刘妍[1] 王刚[1] 施双双[1] 李克[1] 邵得志[1] 斯崇文[2] 

机构地区：[1]解放军第三○二医院传染病研究所基因治疗研究中心,北京100039 [2]北京大学第一医院感染疾病科

出　　处：《中华传染病杂志》2002年第3期148-151,共4页Chinese Journal of Infectious Diseases

基　　金：全军"九五"公关课题资助项目 ( 98D0 6 3);教育部博士点基金资助项目 ( 992 4 )

摘　　要：目的　构建HBV囊膜中蛋白核酸疫苗表达载体并免疫小鼠 ,观察白细胞介素 18(IL 18)对基因免疫的辅助作用。方法　构建质粒 pVR10 12 M、pcDNA 3 .1- IL 18,肌内注射法免疫 2 5只Balb/c小鼠 ,3组小鼠分别注射 10 0 μgpVR10 12 ,pVR10 12 M ,pVR10 12 M和 pcDNA 3 .1- IL 18质粒 ,每 2周 1次 ,共 3次。每次免疫 2周后眼眶采血 ,检测血清抗 HBs,第 3次免疫后 2周应用乳酸脱氢酶检测法验证特异性细胞杀伤率。结果　经注射上述质粒后 ,可观察到注射 pVR10 12 M组的小鼠随免疫次数的增加 ,抗 HBs阳性率、抗体滴度均逐步增高 ;同时联用质粒 pcDNA 3 .1- IL 18的小鼠抗 HBs阳性率、抗体滴度均较单独应用 pVR10 12 M组为低 (第 3次免疫后抗 HBs阳性率、抗体滴度两组比较P <0 .0 5 )。细胞毒性实验证实联用组的细胞杀伤率为 (89.0 2± 15 .5 4) % ,较单独应用pVR10 12 M组 (83 .0 8± 14 .0 2 ) %为高 ,但两组之间差异无显著性。 结论　注射质粒 pVR10 12 M可诱导小鼠产生足量的抗 HBs ,并可检测出特异性细胞免疫反应 ;联合应用IL 18对特异性体液免疫有抑制作用 。Objective To construct plasmid pVR1012 M as nuclei acid vaccine for hepatitis B,was constructed to immunize mice with or without plasmid pcDNA 3.1 - IL 18 to identify the effect of Interleukin 18(IL 18). Methods Polymerase chain reaction method was used to amplify the PreS2 and S region of HBV and reconstruct plasmid pVR1012 M as nuclei acid vaccine. Plasmid pcDNA 3.1 - IL 18 was used as a co stimulator. Twenty five Balb/c mice were divided into 3 groups, group 1 immunized with 100 μg plasmid pVR1012 group 2 pVR1012 M,Group 3,pVR1012 M with pcDNA 3.1 - IL 18, respectively, every 2 weeks for 3 times. Anti HBs were detected in serum 2 weeks after each injection. Lactated ehydrogenase (LDH) cytotoxicity assay was done to analyze the cytotoxic T lymphocytes funciton. Results The positive rate and the antibody titer of serum from mice injected pVR1012 M increasing gradually with the increasing frequency of inoculation, while those from mice injected pVR1012 M and pcDNA 3.1 - IL 18(joint group) were lower than those injected with pVR1012 M alone (Difference after 3rd inoculation was significant, P<0.05) . LDH assay results indicated that the cytotoxicity percent of joint group was higher than that of pVR1012 M group, but without significant difference. Conclusions pVR1012 M can induce both humoral and celluar immunity response specific for HBV in mice. IL 18 may down regulate the humoral and upregulate the cellular immunity response.

关 键 词：乙型肝炎病毒 囊膜 白细胞介素18 基因免疫 蛋白核酸疫苗 动物实验 

分 类 号：R392.1[医药卫生—免疫学]