携带禽流感病毒DNA疫苗的减毒鼠伤寒沙门菌的免疫效力  

Immune efficacy of the attenuated Salmonella typhimurium harbouring DNA vaccine against avian influenza virus

作　　者：潘志明[1] 唐丽华[1] 程宁宁[1] 黄金林[1] 胡青海[1] 焦新安[1] 刘秀梵[1] 

机构地区：[1]扬州大学江苏省人兽共患病学重点实验室,江苏扬州225009

出　　处：《中国兽医科学》2007年第2期125-130,共6页Chinese Veterinary Science

基　　金：国家自然科学基金项目(30425031);全国优秀博士学位论文作者专项资金资助项目(FANEDD200358);江苏省自然科学基金项目(BK2003043);扬州大学"新世纪人才工程"优秀青年骨干教师资助项目(2004年度)

摘　　要：将从重组质粒pVAX1-HA中扩增的HA基因和从pVAX1-HA-CpG中切出的HA-CpG基因分别克隆入pcDNA3.1(+)载体中,获得重组质粒pcDNA3.1-HA和pcDNA3.1-HA-CpG。将上述重组质粒分别转染COS-7细胞,经间接免疫荧光试验检测发现HA基因可在COS-7细胞中表达。将重组质粒转入减毒鼠伤寒沙门菌SL7207,构建成携带DNA疫苗的重组沙门菌SL7207(pcDNA3.1-HA)和SL7207(pcDNA3.1-HA-CpG)。将重组菌分别以5×109CFU口服免疫1日龄鸡2次,重组菌免疫组产生的小肠黏膜抗体效价与空载体组及油乳剂灭活疫苗组间存在显著差异。攻毒免疫保护结果表明,SL7207(pcDNA3.1-HA-CpG)免疫组与空载体组之间存在显著差异,而SL7207(pcDNA3.1-HA)免疫组则与空载体组间无显著差异,说明CpG可增强DNA疫苗的免疫效果。The hemagglutinin protein（HA） gene of avian influenza virus was amplified from pVAX1- HA by PCR and sub-cloned into pcDNA 3.1（＋） vector. HA-CpG gene was also cloned into the pcDNA 3. 1（q-）. The two recombinant plasmids were transfected respectively into COS-7 cells, and identified for the transient expression of HA gene by indirect immunofluorescent assay. Finally, the two recombinant plasmids were transformed into the attenuated Salmonella typhimurium SL7207, and the recombinants were designated as SL7207（pcDNA3. 1-HA） and SL7207（pcDNA3. 1-HA-CpG）, respectively. One-day-old commercial ISA brown chickens were immunized with one of the 2 recombinant bacteria at 5 × 10^9 CFU, respectively and boosted with the same dose two weeks later. No significant difference was observed in body weight among those groups. Intestinal mucosal immune response was observed in chickens immunized with one of the 2 recombinant bacteria on day 35, and their levels were significantly higher than those of the negative and positive controls. Compared with the negative control group, protective immunity induced by SL7207（pcDNA3.1-HA-CpG） provided significant protection, but SL7207（pcDNA3.1-HA） did not, indicating that the combination of CpG can enhance the immune response of the DNA vaccine delivered by the attenuated Salmonella.

关 键 词：禽流感病毒 减毒鼠伤寒沙门菌 DNA疫苗 CPG 免疫效力 

分 类 号：S852.659.5[农业科学—基础兽医学]