泰山松花粉多糖对PRRSV GP5亚单位疫苗免疫增强作用的研究  

Research on immunological enhancement effect of Taishan Pinus massoniana pollen polysaccharides on subunit vaccine of porcine reproductive and respiratory syndrome virus glycoprotein 5(GP5)

作　　者：袁艳梅 彭军[1] 胡莉萍[2] 朱丽君[1] 王淑娟[1] 董雯雯[1] 马宁宁[1] 周建波[1] 卢亚平 李宗瑞 魏凯[1] 王海荣[1] 朱瑞良[1] 

机构地区：[1]山东农业大学动物科技学院/山东省动物生物技术与疫病防治重点实验室,山东泰安271018 [2]山东省动物疫病预防与控制中心,山东济南250022

出　　处：《中国预防兽医学报》2017年第9期740-745,共6页Chinese Journal of Preventive Veterinary Medicine

基　　金：十三五国家重点研发计划畜禽重大疫病防控与高效安全养殖综合技术研发重点专项项目(2017YFD0500706);山东省重点研发计划项目(2016GNC110013)

摘　　要：为研究泰山松花粉对猪繁殖与呼吸综合征病毒(PRRSV)GP5的免疫增强作用,本研究以原核表达的高致病性PRRSV GP5蛋白为抗原,添加不同剂量的天然提取的泰山松花粉多糖(TPPPS)作为免疫佐剂配制PRRSV亚单位疫苗。选取72只SPF昆明小鼠随机分为6组(Ⅰ-Ⅵ),12只/组。Ⅰ组免疫纯化的GP5亚单位疫苗,Ⅱ-Ⅳ组分别免疫不同浓度的TPPPS(20 mg/mL、40 mg/mL、60 mg/mL)佐剂GP5亚单位疫苗,V组免疫弗氏不完全佐剂GP5亚单位疫苗,VI组注射PBS;1周后各组均加强免疫1次,加强免疫后,每周每组随机挑选3只小鼠采集外周血,分别检测小鼠血清抗体效价、T淋巴细胞亚群比例、T淋巴细胞转化率和细胞因子的含量,评价TPPPS对PRRSV-GP5蛋白诱导的免疫应答反应的调节作用。结果显示,所有疫苗免疫组小鼠的上述免疫指标比对照组均有显著提高,而添加TPPPS和弗氏佐剂的疫苗组各项指标均显著高于单独GP5蛋白免疫组。3个浓度的TPPPS佐剂中,60 mg/mL浓度的免疫增强效果最显著。本研究表明,重组表达的PRRSV GP5蛋白能够诱导动物机体显著的免疫应答,TPPPS佐剂对PRRSV GP5蛋白呈现显著的免疫增强效果。本研究为研制PRRSV亚单位疫苗提供了参考。To evaluate the immunomodulation effects of Taishan Pinus massoniana pollen polysaccharide （TPPPS） used as adjuvant on a subunit vaccine prepared by glycoprotein 5 （GPS） which was expressed through prokaryotic expression system 72 SPF Kunming mice （weight of about 20 g） were divided into 6 groups （I-VI） randomly. Each mouse in group I-VI was immunized with purified GP5 subunit vaccine, 20 mg/mL, 40 mg/mL, and 60 mg/mL of TPPPS adjuvant GP5 subunit vaccines, the Freund＇s incomplete adjuvant GP5 subunit vaccine, and PBS respectively. Booster immunization was conducted at 7 days post the first immunization. The titers of serum antibody, percentages of T lymphocyte, lymphocyte transformation rate, concentrations of cytokines of the mice in different groups were tested. The results showed that all immune indexes of mice in Freund＇s adjuvant group and TPPPS adjuvant groups were higher than the pured GP5 subunit vaccine group. Among three concentrations of TPPPS, 60 mg/mL TPPPS showed optimal effects. Our findings indicated that TPPPS adjuvant showed significant immune enhancement effect on recombinant PRRSV GP5 protein, and PRRSV GP5 protein could induce strong immune response. This study provided significant reference for the research of PRRSV subunit vaccine which is of better ability of inducing neutralizing protection effect.

关 键 词：泰山松花粉多糖 免疫增强作用 猪繁殖与呼吸综合征病毒 GP5蛋白 亚单位疫苗 

分 类 号：S852.61[农业科学—基础兽医学]