传染性喉气管炎病毒基因疫苗免疫效应  

Immune Efficacy of DNA Vaccine of Avian Infectious Laryngotracheitis Virus

作　　者：王利丽[1] 王云峰[1] 孙惠玲[1] 杨春富[1] 都兴洋[1] 王玫[1] 步志高[1] 童光志[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室禽传染病研究室

出　　处：《畜牧兽医学报》2007年第2期175-183,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家"863"计划资助项目(2003AA213021)

摘　　要：以传染性喉气管炎病毒(ILTV)WG株gB和gD基因为目的基因构建真核表达质粒pCAGG-gB和pCAGG-gD。将75只SPF鸡随机分为7组,经肌肉注射接种pCAGG-gB、pCAGG-gD、pCAGG-gB+pCAGG-gD、pCAGG-gB+rFPV-ILTVgB以及rFPV-ILTVgB,并设有生理盐水和空载体(pCAGG)对照组。免疫2周后以500EID50ILTV WG株强毒进行攻击,以评价ILTV基因疫苗以及基因疫苗和重组病毒联合应用对SPF鸡的免疫保护作用。血清抗体的检测表明:在免疫后的2周,各免疫组的鸡只均产生了特异性的ILTV抗体,外周血CD4+T、CD8+T、TCRγδ+T细胞明显高于对照组,并且pCAGG-gB和pCAGG-gD分别诱导了高水平的IL-18和IFN-γ的表达。ILTV强毒攻击后,非免疫鸡全部发病,并在第2天出现死亡。而各质粒免疫组均获得了一定的保护(保护率44%),两种质粒联合免疫(pCAGG-gB和pCAGG-gD)和重组病毒(rFPV-ILTVgB)与质粒(pCAGG-gB)联合免疫的免疫组的保护率(56%)高于单独免疫组,产生高水平细胞因子表达的免疫组的保护率高于对照组。重组鸡痘病毒免疫组的保护率(69%)最高。这些结果表明ILTV的DNA疫苗能够诱导鸡体产生特异性免疫应答,并能对传染性喉气管炎强毒的攻击提供一定的免疫保护。The gB and gD gene of Infectious Laryngotracheitis virus （ILTV） WangGang （WG） strain were cloned into the eukaryotic expression vector pCAGGS and designated as pCAGG-gB, pCAGG-gD, respectively. Seventy-five 30-day-old SPF white Leghorn chickens were randomly di- vided into seven groups , five groups of them were immunized by four sites intramuscular injection with pCAGG-gB, pCAGG-gD, pCAGG-gB＋pCAGG-gD, pCAGG-gB＋rFPV-ILTVgB, rFPV-IL-TVgB, respectively. Another two groups were injected with sodium chloride solution and pCAGGS as control groups. Two weeks after the second immunization, chickens were challenged intralaryngealy with 500EID50 of highly virulent ILTV WG strain. The sera antibodies detection results showed that all chickens immunized with pCAGG-gB, pCAGG-gD, pCAGG-gB＋pCAGG-gD, pCAGG-gB ＋ rFPV-ILTVgB, rFPV-ILTVgB developed ILTV-specific antibodies 2 weeks post-inoculation. The percentages of CD4^＋ , CD8^＋ and TCRγδ^＋ T lymphocytes of these immunized groups were higher than that of control groups, and expression level of cytokine IFN-γ and IL-18 were higher in pCAGG-gB and pCAGG-gD groups than control groups. After challenge, chickens in control groups began to die at day 2 post-challenge. Chickens immunized with pCAGG-gB, pCAGG-gD had been partially protected （44%）, the group immunized with two plasmids （pCAGG-gB and pCAGG-gD） and the group immunized with plasmid associated with recombinant virus （pCAGG-gB ＋ rFPV-ILTVgB） had higher protection （56%）, the group immunized with recombinant virus （rFPV-ILTVgB） had the highest protection （69%）. These results indicated that DNA immunization can induce immune response and partially protect chickens from homologous ILT virus challenge.

关 键 词：鸡传染性喉气管炎病毒(ILTV) DNA疫苗 免疫效力 

分 类 号：S852.65[农业科学—基础兽医学]