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猪源共表达衣壳前体蛋白(P1)和口蹄疫病毒多表位肽的重组伪狂犬病病毒的构建及免疫反应特性分析

Construction and immune response characterization of a recombinant pseudorabies virus co-expressing capsid precursor protein (P1) and a multiepitope peptide of foot-and-mouth disease virus in swine.

作者信息

He Yannan, Qian Ping, Zhang Keshan, Yao Qingxia, Wang Dang, Xu Zhuofei, Wu Bin, Jin Meilin, Xiao Shaobo, Chen Huanchun

机构信息

State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, Hubei 430070, PR China.

出版信息

Virus Genes. 2008 Apr;36(2):393-400. doi: 10.1007/s11262-008-0204-6. Epub 2008 Feb 2.

Abstract

Foot-and-mouth disease (FMD) is the most contagious and devastating disease of livestock. Our previous studies demonstrated that TK(-)/gG(-)/P1(+), a recombinant expressing the FMDV capsid precursor protein (P1) based on attenuated pseudorabies virus (PRV) TK(-)/gG(-), could be used as a recombinant vaccine to protect pigs against both pseudorabies and FMD. However, because the P1 expression cassette is inserted into the gG locus of the genome of PRV TK(-)/gG(-), this bivalent vaccine cannot be used in conjunction with the PRV gE-ELISA, an extensively used discriminatory serological test in eradication programs for pseudorabies, which limits the clinical use of this bivalent vaccine. To circumvent this shortcoming, in this study, an expression cassette containing synthetic multiepitope gene "FHG" consisting of six potential B cell epitopes and two potential T cell epitopes of FMDV, under the control of CMV promoter, was further inserted into the gE/gI locus of genome of TK(-)/gG(-)/P1(+), resulting in a new recombinant FHG/P1/PRV. The immunogenicity of FHG/P1/PRV was evaluated and compared with TK(-)/gG(-)/P1(+) in piglets. Our results clearly showed that FHG/P1/PRV performed better than or comparable with TK(-)/gG(-)/P1(+), as demonstrated by comparable PRV-specific neutralizing antibodies, enhanced FMDV-specific neutralizing antibodies, and cellular immune responses. More importantly, no gE- and gG-specific antibodies could be detected in pigs immunized with FHG/P1/PRV. These data indicate that FHG/P1/PRV is a promising bivalent vaccine candidate with more extensive potential application than TK(-)/gG(-)/P1(+) against both pseudorabies and FMD.

摘要

口蹄疫(FMD)是家畜中传染性最强、危害最大的疾病。我们之前的研究表明,基于减毒伪狂犬病病毒(PRV)TK(-)/gG(-)构建的表达口蹄疫病毒(FMDV)衣壳前体蛋白(P1)的重组体TK(-)/gG(-)/P1(+),可作为重组疫苗用于保护猪免受伪狂犬病和口蹄疫的侵害。然而,由于P1表达盒插入到了PRV TK(-)/gG(-)基因组的gG位点,这种二价疫苗不能与PRV gE-ELISA联合使用,而PRV gE-ELISA是伪狂犬病根除计划中广泛使用的鉴别性血清学检测方法,这限制了这种二价疫苗的临床应用。为了克服这一缺点,在本研究中,一个包含由FMDV的六个潜在B细胞表位和两个潜在T细胞表位组成的合成多表位基因“FHG”的表达盒,在CMV启动子的控制下,进一步插入到TK(-)/gG(-)/P1(+)基因组的gE/gI位点,产生了一种新的重组体FHG/P1/PRV。对FHG/P1/PRV的免疫原性进行了评估,并与仔猪中的TK(-)/gG(-)/P1(+)进行了比较。我们的结果清楚地表明,FHG/P1/PRV的表现优于TK(-)/gG(-)/P1(+)或与之相当,表现为相当的PRV特异性中和抗体、增强的FMDV特异性中和抗体以及细胞免疫反应。更重要的是,在用FHG/P1/PRV免疫的猪中未检测到gE和gG特异性抗体。这些数据表明,FHG/P1/PRV是一种有前景的二价疫苗候选物,与TK(-)/gG(-)/P1(+)相比,在抗伪狂犬病和口蹄疫方面具有更广泛的潜在应用。

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