传染性法氏囊病病毒单克隆抗体的制备及其抗病毒活性的分析  

Preparation of the monoclonal antibodies against infectious bursal disease virus and their antiviral activities

作　　者：夏兴霞[1] 欧阳伟[1] 王永山[1] 诸玉梅[1] 潘群兴[1] 毕振威[1] 王晓丽[1] 吴晓悠[1] 董晨红[1] 

机构地区：[1]江苏省农业科学院兽医研究所农业部兽用生物制品工程技术重点实验室国家兽用生物制品工程技术研究中心/江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009

出　　处：《中国兽医科学》2014年第3期277-281,共5页Chinese Veterinary Science

基　　金：国家自然科学基金项目(31272537);江苏省自然科学基金项目(BK2010471;BK2012787);江苏省农业科技自主创新资金项目[CX(13)5034]

摘　　要：以传染性法氏囊病病毒(IBDV)病毒样颗粒重组VP2蛋白免疫BALB/c小鼠,制备免疫脾细胞,与SP2/0骨髓瘤细胞融合,获得7株分泌抗IBDV单克隆抗体(McAb)的杂交瘤细胞。用间接免疫荧光试验和病毒中和试验分别测定McAb的特异性和抗病毒活性,杂交瘤细胞株2C8分泌的McAb具有较高的抗IBDV活性,培养上清液中的抗体中和效价为210,诱生BALB/c小鼠腹水中的抗体中和效价达到107,抗体亚类为IgG1κ。将2C8-McAb制成注射液,经肌肉注射,在12h内,鸡血液循环中的鼠源McAb达到高峰,半衰期至少可持续14d。在IBD预防试验中,注射2C8-McAb的鸡可抵抗IBDV强毒感染。在IBD治疗试验中,对IBDV强毒感染发病鸡用2C8-McAb治疗,存活率为60%(18/30),第21天检测法氏囊组织IBDV全部为阴性;而非治疗对照组,发病鸡的存活率只有20%(6/30),第21天检测存活鸡法氏囊组织IBDV全部呈阳性。本试验结果表明,高中和活性的2C8-McAb对IBD具有良好的预防及治疗效果,能够显著降低死亡率,并且McAb治疗康复鸡不带毒。Seven hybridomas secreting monoclonal antibodies（McAb） against VP2 of infectious bursal disease virus（IBDV） were established by fusing SP2/0 with spleen cells from BALB/c mice immunized with virus-like particles being recombinant VP2 protein. The specificity and antiviral activity of the McAbs were identified using indirect immunofluoresence assay and virus neutralization test, respectively. The clone 2C8 was the best with the neutralization titer of 2^10 and 107 in the cultural supernatant and the ascitic fluid, respectively. The subtype of its immunoglobulin was IgG1κ When 2C8-McAb was intramuscularly injected into chicken,the titer of 2C8-McAb in the blood circulation reached the peak at hour 12 post-injection and its half-life could persist 14 d. The chicken injected with 2C8-McAb could effectively prevent against challenge of virulent IBDV. As the chicken infected with virulent IBDV were treated with the 2C8-McAb, the survival rate of the infected fowl was 60% （18/30）,and the survival chickens were all negative in the virus detection of the bursal tissues after 21 d. However,the control group with the survival rate was only 20~//00（6/30）,and IBDV were positive. The results ind significantly preventive and therapeutic effectiveness on IBD. out 2C8-McAb treatment, icated that 2C8-McAb had

关 键 词：传染性法氏囊病病毒(IBDV) 单克隆抗体(McAb) 抗病毒活性 

分 类 号：S852.659.4[农业科学—基础兽医学]