[1]石晓玉,张元鹏,乔绪稳,等.重组脑膜炎球菌PorB蛋白对猪伪狂犬弱毒疫苗的免疫增强效力[J].江苏农业学报,2017,(02):353-360.[doi:doi:10.3969/j.issn.1000-4440.2017.02.018]
　SHI Xiao-yu,ZHANG Yuan-peng,QIAO Xu-wen,et al.Immune enhancement to porcine pseudorabies virus(PRV) vaccine of recombinant Neisseria meningitides PorB protein[J].,2017,(02):353-360.[doi:doi:10.3969/j.issn.1000-4440.2017.02.018]
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重组脑膜炎球菌PorB蛋白对猪伪狂犬弱毒疫苗的免疫增强效力()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2017年02期
页码:
353-360
栏目:
畜牧兽医·水产养殖
出版日期:
2017-04-30

文章信息/Info

Title:
Immune enhancement to porcine pseudorabies virus(PRV) vaccine of recombinant Neisseria meningitides PorB protein
作者:
石晓玉12; 张元鹏2; 乔绪稳2; 于晓明2; 陈瑾2; 姜平1; 郑其升2; 侯继波2
(1.南京农业大学动物医学院，江苏南京210095；2.国家兽用生物制品工程技术研究中心，江苏南京210014）
Author(s):
SHI Xiao-yu12; ZHANG Yuan-peng2; QIAO Xu-wen2; YU Xiao-ming2; CHEN Jin2; JIANG Ping1; ZHENG Qi-sheng2; HOU Ji-bo2
（1.College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China;2.National Veterinary Biological Medicine Engineering Research Center, Nanjing 210014, China）
关键词:
脑膜炎球菌PorB蛋白; 猪伪狂犬弱毒疫苗; 免疫增强
Keywords:
recombinant PorB; pseudorabies virus(PRV) attenuated vaccine; immune-enhancing activity
分类号:
S852.4
DOI:
doi:10.3969/j.issn.1000-4440.2017.02.018
文献标志码:
A
摘要:
探讨原核表达的重组脑膜炎球菌外膜蛋白PorB对猪伪狂犬弱毒活疫苗免疫的增强效果。将GenBank公布的脑膜炎球菌外膜蛋白PorB基因克隆到原核表达载体pQZ Ⅲ中，命名为pQZ-PorB质粒，转入大肠杆菌BL21中表达重组蛋白。采用SDS-PAGE、Western-blot技术和Ni2+-NTA柱鉴定并纯化重组蛋白。将每份含100 μg剂量的重组PorB蛋白与猪伪狂犬弱毒疫苗混合后分别滴鼻免疫3日龄仔猪和肌肉注射免疫50日龄猪，同时设PBS对照组和免疫前对照组。免疫28 d后测定IgG和IgA抗体水平、猪外周血淋巴细胞增殖情况以及细胞因子（IFN-γ、IL-10、IL-6）的表达水平。SDS-PAGE和Western-blot结果表明，PorB基因在大肠杆菌中表达，表达的重组PorB蛋白是可溶性的，Ni2+-NTA柱能够有效纯化重组蛋白。重组PorB蛋白不但能够刺激免疫猪在较短时间内产生的较高血清IgG抗体、黏膜IgA抗体和中和抗体，而且可以通过诱导机体产生IFN-γ和IL-6来促进细胞免疫反应。在猪伪狂犬弱毒疫苗免疫中，重组PorB蛋白可以作为一种优良的免疫增强剂。
Abstract:
In order to explore the immune-enchancing activity to swine pseudorabies virus(PRV) attenuated vaccine of Neisseria meningitides PorB, Escherichia coli codon-optimized mature PorB gene was synthetized and cloned into the prokaryotic expression vector pQZ III. The recombinant PorB protein was expressed in E. coli BL21(DE3). The recombinant protein was confirmed and purified by SDS-PAGE, Western blot and Ni-NTA affinity chromatography. The PorB protein (100 μg) was mixed with live attenuated PRV vaccine, and used to immunize intranasally 3-day-aged piglets and intramuscularly 50-day-aged pig, respectively. PBS and pre-immune control group were set. The levels of IgG, IgA , peripheral blood lymphocyte proliferation and cytokines were detected at 28 d postimmunization. The recombinant protein PorB not only stimulated swines to produce high levels of serum IgG antibody, mucosal IgA antibody and neutralizing antibody in a relatively short period of time, but also facilitated immune responces by inducing the production of IFN-γ and IL-6. Results indicate that recombinant PorB may be effective as an adjuvant with PRV attenuated vaccines.

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备注/Memo

备注/Memo:
收稿日期：2016-07-25 基金项目：公益性行业(农业)科研专项(201303046)；江苏省农业自主创新专项[CX(14)2089] 作者简介：石晓玉（1990-），女，河南濮阳人，硕士研究生，主要从事新型猪用疫苗研究。（Tel）15951979330;（E-mail）1334318532@qq.com 通讯作者：姜平，（Tel）025-84395504；（E-mail）jiangp@njau.edu.cn。郑其升，（Tel）025-84392088；（E-mail）njcvc1302@163.co
更新日期/Last Update: 2017-05-02