鸡传染性支气管炎病毒S1基因和鸡II型干扰素基因在重组鸡痘病毒中的共表达  

Co-expression of S1 Gene of Infectious Bronchitis Virus and Chicken Type II Interferon Gene in Recombinant Fowlpox Virus

作　　者：孙永科[1] 田占成[1] 王云峰[1] 童光志[1] 智海东[1] 刘胜旺[1] 王玫[1] 杨增岐[2] 

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,哈尔滨150001 [2]西北农林科技大学动物科技学院,杨凌712100

出　　处：《畜牧兽医学报》2005年第8期800-806,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家"863"计划资助项目(2003AA213020)

摘　　要：将鸡II型干扰素(ChIFNγ)基因和鸡传染性支气管炎病毒S1基因同时插入到鸡痘病毒转移载体中,构建含有这2个基因的鸡痘病毒转移载体pSY-ChIFNγS1。采用脂质体法将该质粒转染鸡痘病毒感染的鸡胚成纤维细胞(CEF),经过8轮蓝斑筛选,得到纯化的能够同时表达鸡II型干扰素和鸡传染性支气管炎病毒S1基因的重组鸡痘病毒rFPV-ChIFNγS1。rFPV-ChIFNγS1接种28日龄的SPF鸡1周后,可以检测到针对传染性支气管炎病毒S1基因的ELISA抗体,重组病毒接种鸡的CD4+、CD8+和γδTCR阳性T淋巴细胞的百分比含量显著高于非免疫对照鸡(P<0.05);免疫4周后用传染性支气管炎LX4强毒株攻击,rFPV-ChIFNγS1免疫组仅有1只出现轻微的呼吸道症状(1/16),而非免疫对照组则有16只发病(16/16),并有2只出现死亡(2/16),表明rFPV-ChIFNγS1对接种鸡可以产生良好的保护效果。In this study, the chicken type Ⅱ interferon gene（ChIFNγ） and infectious bronchitis virus （IBV） S1 gene were inserted into fowlpox virus transferring vector, for constructing pSY-ChIFNγS1 which contained these two genes. Then by lipofectin, the pSY-ChIFNγS1 was transfected into the chicken embryo fibroblasts （CEF） which was pre-infected with wide type fowlpox virus. After eight times selections of blue plagues on the CEF overlaid using X-gal as substrate, we obtained the purified rFPV-ChIFNγS1 which could express the S1 and ChIFNγ, protein at the same time. Special antibody to IBV S1 protein can be identified by ELISA about one week after immunization of 28 days SPF chickens with rFPV-ChIFNγS1. The percentages of CD4^＋ , CD8^＋ and γδTCR positive T lymphocyte in peripheral blood of immune group are remarkably higher than that of Mock. Four weeks after immunization, the two groups were challenged with violent IBV LX4 strain, and only 1/16 chicken appeared light respiratory symptom in rFPV-ChIFNγS1 immunized group, but 16/16 chickens appeared in Mock, and 2/16 chickens died. The result of animal trail showed, rFPV-ChIFNγS1 can protect immunized chickens well.

关 键 词：传染性支气管炎病毒 鸡Ⅱ型干扰素 重组鸡痘病毒 

分 类 号：S852.65[农业科学—基础兽医学] Q784[农业科学—兽医学]