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当前位置: 首页 > 文章 > 牡丹籽蛋白酶解产物对禽多杀性巴氏杆菌ptfA基因DNA疫苗免疫效果的影响 中国预防兽医学报 2019,41 (1) 1053-1058
Position: Home > Articles > The effect of peony seed proteolysis product on the immune efficacy of ptfA gene DNA vaccine of avian Pasteurellamultocida Chinese Journal of Preventive Veterinary Medicine 2019,41 (1) 1053-1058

牡丹籽蛋白酶解产物对禽多杀性巴氏杆菌ptfA基因DNA疫苗免疫效果的影响

作  者：
杜珍奇;宫强;牛明福;秦翠丽;任国艳;马丽苹;伍家发;张彬
单  位：
河南科技大学 食品与生物工程学院/食品微生物河南省工程技术研究中心
关键词：
禽多杀性巴氏杆菌;牡丹籽蛋白酶解产物;ptfA基因DNA疫苗;免疫效果
摘  要：
为研究牡丹籽蛋白酶解产物对禽多杀性巴氏杆菌(Pm)ptfA基因DNA疫苗免疫效果的影响,本研究以牡丹籽蛋白酶解产物为佐剂制备佐剂-DNA疫苗,分别以DNA疫苗和佐剂-DNA疫苗免疫鸡,同时设置牡丹籽蛋白酶解产物灌胃-DNA疫苗免疫组以及弱毒疫苗、pcDNA3.1(+)空载体和PBS对照组.每两周免疫一次,共免疫3次.间接ELISA检测免疫后鸡血清特异性抗体水平,MTT法检测免疫鸡外周血淋巴细胞增殖情况,双抗夹心ELISA检测鸡IL-2和IL-4的分泌情况.三免2周后以强毒菌株CVCC474(5LD50/只)攻毒,计算强毒攻击后各组鸡的存活数及保护率.结果显示,佐剂-DNA疫苗组和灌胃组鸡的血清特异性抗体水平与DNA疫苗组相比无显著差异(p>0.05).三免后,佐剂-DNA疫苗组和灌胃组鸡的SI值和IL-4水平显著高于DNA疫苗组(p<0.05).二免和三免后,佐剂-DNA疫苗组和灌胃组鸡的IL-2水平明显高于DNA疫苗组(p<0.05),且为免疫鸡提供的保护率高于DNA疫苗,其中以牡丹籽蛋白酶解产物为佐剂时效果较好,但均不及弱毒疫苗.表明预先灌胃牡丹籽蛋白酶解产物或以其为佐剂均可在一定程度上增强ptfA基因DNA疫苗的免疫效果.本研究为PmDNA疫苗的研究及牡丹籽蛋白酶解产物在疫苗佐剂中的应用提供一定的参考.
译  名：
The effect of peony seed proteolysis product on the immune efficacy of ptfA gene DNA vaccine of avian Pasteurellamultocida
作  者：
DU Zhen-qi;GONG Qiang;NIU Ming-fu;QIN Cui-li;REN Guo-yan;MA Li-ping;WU Jia-fa;ZHANG Bin;College of Food and Bioengineering/Henan Engineering and Technology Research Center of Food Microbiology,Henan University of Science and Technology;
关键词：
avian Pasteurella multocida;;peony seed proteolysis produc;;ptf A gene DNA vaccine;;immune efficacy
摘  要：
To study the effect of peony seed proteolysis product on the immune efficacy of ptfA gene DNA vaccine of avian Pasteurella multocida, the adjuvant-DNA vaccine gene was prepared with peony seed proteolysis product as adjuvant. To evaluate the immune efficacy of adjuvant-DNA vaccine, chickens were vaccinated with DNA vaccine, adjuvant-DNA vaccine, with and peony seed proteolysis by intragastric administration plus DNA vaccine(gavage group), respectively. Chickens in control group were vaccinated with attenuated live vaccine, pcDNA 3.1(+) empty vector and PBS, respectively. All groups were boosted twice,and each vaccination is in two weeks interval. The serum antibodies were detected by indirect ELISA. The peripheral blood lymphocyte(PBL) proliferation was tested by MTT. IL-2 and IL-4 secreted by PBL were detected by double-antibodies-sandwichELISA. Chickens were challenged with avian Pasteurella multocida on week 2 post the third immunization, the survival number and protection rate were counted. The results of ELISA showed there were no difference in the antibody levels among the adjuvant-DNA vaccine group, gavage group and DNA vaccine groups(p>0.05). The SI values and levels of IL-4 in adjuvant-DNA vaccine group and gavage group were significantly higher than that of DNA vaccine groups after the third immunization(p<0.05).The levels of Il-2 secreted by PBL in the adjuvant-DNA vaccine group and gavage group were significantly higher than that of DNA vaccine groups after the second and the third immunization(p<0.05). The protection rate of adjuvant-DNA vaccine group and gavage group were higher than that of the DNA vaccine group, especially the adjuvant-DNA vaccine was better. These results indicated the immune efficacy of ptfA gene DNA vaccine could be enhanced by pre-administration of peony seed proteolysis product or using it as adjuvant. These findings will to lay the foundation for the development of DNA vaccine against avian Pasteurella multocida and the application of peony seed proteolysis product in the vaccine as adjuvant.

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