丙型肝炎病毒内源性靶向基因疫苗对荷瘤小鼠抑瘤效果的初步研究  

A study on the inhibitory effect on transplanted tumor growth expressing HCV NS3 protein by HCV minigene vaccine based on invariant chain substitution in mice

作　　者：高明 王海平 周勇 王嘉军 王燕宁[2] 王全立 

机构地区：[1]输血医学研究所,北京100850 [2]兰州军区兰州总医院输血科

出　　处：《中华肝脏病杂志》2005年第11期801-804,共4页Chinese Journal of Hepatology

基　　金：国家自然科学基金(30200245)

摘　　要：目的研究基于恒定链的丙型肝炎病毒(HCV)NS3内源性靶向巨噬细胞表面分子Ia的分子基因疫苗对HCV NS3荷瘤小鼠的抑瘤效果,探索HCV感染基因治疗的可行途径。方法BALB/c小鼠皮下接种稳定表达HCV NS3的小鼠骨髓瘤SP2/0-NS3细胞,3 d后于股四头肌注射疫苗进行免疫治疗,2周后进行一次加强免疫。BALB/c小鼠共64只,随机分为等渗盐水对照组、空质粒pCI-neo对照组、pHCV- NS3非靶向治疗组和pHCV-NS3-Thl靶向治疗组,观察成瘤时间、肿瘤大小和小鼠的60 d存活率。结果等渗盐水、pCI-neo、pHCV-NS3、pHCV-NS3-Th1组小鼠平均成瘤时间分别为(16.17±2.55)d、(14.40 ±1.82)d、(16.75±2.36)d、(24.00±5.57)d;成瘤率分别为100.0%(13/13)、100.0%(14/14)、57.1%(8/14)和46.7%(7/15);60 d存活率分别为0、0、50.0%(7/14)和53.3%(8/15)。对四个时间点成瘤小鼠肿瘤大小比较的结果显示,pHCV-NS3-Th1组成瘤小鼠的肿瘤直径最小,与其他组相比,差异均有统计学意义(P<0.01)。结果显示,成瘤小鼠的平均存活时间比等渗盐水组延长6 d,比pCI-neo组延长12 d(P<0.05),比pHCV-NS3组延长6 d。结论pHCV-NS3-Th1具有延迟HCV-NS3荷瘤小鼠肿瘤形成、限制肿瘤生长、降低成瘤率、病死率以及提高存活率的作用。基于恒定链的内源性靶向性基因疫苗有望作为HCV感染防治的候选治疗性疫苗。Objectives To establish a transplanted tumor producing HCV NS3 protein in mice and study the therapeutic effect of minigene vaccine based on invariant chain substitution. Methods SP2/0-NS3 cells expressing HCV NS3 antigen were injected subcutaneously into BALB/c mice. After three days of inoculation, different therapeutical reagents were injected intramuscularly into different groups of mice. The boost immunization was carried out two weeks after the first immunization. The efficiency of HCV NS3 Thl minigene vaccine was estimated after 60 days observation. Results For saline, pCI-neo, pHCV-NS3 and pHCV-NS3-Thl treated groups, the induction period needed for tumor growth was 16.17 ± 2.55, 14.40 ± 1.82, 16.75 ± 2.36, and 24.00 ± 5.57 days （ t =2.623, P =0.034 vs saline, t=3.713, P=0.010 vs pCI-neo and t =2.425, P=0.045 vs pHCV-NS3） respectively. The tumorigenesis rates were 100%, 100%, 57.1% （8/14, χ^2= 6.190, P = 0.013 vs saline and χ^2= 6.608, P = 0.010 vs pCI-neo） and 46.7% （7/15, χ^2 = 9.707, P = 0.002 vs saline and χ^2 = 10.311, P = 0.001 vs pCI-neo ） respectively. The survival rates were 0, 0, 50.0% （7/14, χ^2 = 5.787, P = 0.016 vs saline and χ^2= 9.333, P = 0.002 vs pCI-neo） and 53.3% （8/15, χ^2= 6.651, P = 0.010 vs saline and χ^2= 10.311, P = 0.001 vs pCI-neo） respectively. The average tumor diameter of the pHCV-NS3-Thl treated group was significantlysmaller compared with the control groups and the pHCV-NS3 treated group （P =0.001）. Moreover, the average survival time of tumor-bearing mice immunized with pHCV-NS3-Thl was 6 days longer compared with the saline treated group, 12 days longer compared with the pCI-neo treated group （P =0.001）, and 6 days compared with the pHCV-NS3 treated group. Conclusion HCV NS3 Thl epitope vaccine might be a potential biotherapy candidate against HCV infection.

关 键 词：肝炎病毒 丙型 表位 基因 MHCⅡ类 丙型肝炎病毒 荷瘤小鼠 基因疫苗 抑瘤效果 靶向性 

分 类 号：R392[医药卫生—免疫学]