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融合猪CD154的E2糖蛋白作为新型嵌合亚单位疫苗预防经典猪瘟病毒在怀孕母猪中垂直传播的效果。

Efficacy of E2 glycoprotein fused to porcine CD154 as a novel chimeric subunit vaccine to prevent classical swine fever virus vertical transmission in pregnant sows.

作者信息

Muñoz-González Sara, Sordo Yusmel, Pérez-Simó Marta, Suárez Marisela, Canturri Albert, Rodriguez Maria Pilar, Frías-Lepoureau María Teresa, Domingo Mariano, Estrada Mario Pablo, Ganges Llilianne

机构信息

IRTA, Centre de Recerca en Sanitat Animal (CReSA, IRTA-UAB), Campus de la Universitat Autònoma de Barcelona, 08193 Barcelona, Spain.

Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), Havana, Cuba.

出版信息

Vet Microbiol. 2017 Jun;205:110-116. doi: 10.1016/j.vetmic.2017.05.003. Epub 2017 May 4.

Abstract

Here we evaluated the effect of double vaccination with a novel subunit marker vaccine candidate based in the CSFV E2 glycoprotein fused to the porcine CD154 to prevent CSFV vertical transmission. A lentivirus-based gene delivery system was used to obtain a stable recombinant HEK 293 cell line for the expression of E2 fused to porcine CD154 molecule. Six pregnant sows were distributed in two groups and at 64days of gestation animals numbered 1-4 (group 1) were vaccinated via intramuscular inoculation with 50μg of E2-CD154 subunit vaccine. Animals from group 2 (numbered 5 and 6, control animals) were injected with PBS. Seventeen days later sows from group 1 were boosted with the same vaccine dose. Twenty-seven days after the first immunization, the sows were challenged with a virulent CSFV Margarita strain and clinical signs were registered. Samples were collected during the experiment and at necropsy to evaluate immune response and virological protection. Between 14 and 18days after challenge, the sows were euthanized, the foetuses were obtained and samples of sera and tissues were collected. E2-CD154 vaccinated animals remained clinically healthy until the end of the study; also, no adverse reaction was shown after vaccination. An effective boost effect in the neutralizing antibody response after the second immunization and viral challenge was observed and support the virological protection detected in these animals after vaccination. Protection against CSFV vertical transmission was found in the 100% of serums samples from foetus of vaccinated sows. Only two out of 208 samples (0.96%) were positive with Ct value about 36 corresponding to one tonsil and one thymus, which may be non-infective viral particles. Besides, its DIVA potential and protection from vertical transmission, the novel CSFV E2 bound to CD154 subunit vaccine, is a promising alternative to the live-attenuated vaccine for developing countries.

摘要

在此,我们评估了一种基于猪瘟病毒(CSFV)E2糖蛋白与猪CD154融合的新型亚单位标记疫苗候选物进行双重疫苗接种对预防CSFV垂直传播的效果。使用基于慢病毒的基因递送系统获得稳定的重组人胚肾293(HEK 293)细胞系,用于表达与猪CD154分子融合的E2。六头怀孕母猪分为两组,在妊娠64天时,第1组编号为1 - 4的动物通过肌肉注射接种50μg E2 - CD154亚单位疫苗。第2组(编号为5和6,对照动物)的动物注射磷酸盐缓冲液(PBS)。17天后,第1组母猪用相同剂量的疫苗进行加强免疫。首次免疫27天后,母猪用强毒CSFV玛格丽塔毒株进行攻毒,并记录临床症状。在实验期间和尸检时采集样本,以评估免疫反应和病毒学保护。攻毒后14至18天,对母猪实施安乐死,获取胎儿,并采集血清和组织样本。接种E2 - CD154的动物在研究结束前一直保持临床健康;此外,接种疫苗后未出现不良反应。观察到第二次免疫和病毒攻毒后中和抗体反应有有效的加强作用,并支持在这些动物接种疫苗后检测到的病毒学保护。在接种疫苗母猪所产胎儿的100%血清样本中发现了针对CSFV垂直传播的保护作用。208个样本中只有2个(0.96%)呈阳性,其Ct值约为36,对应一个扁桃体和一个胸腺,这可能是无感染性的病毒颗粒。此外,除了其鉴别诊断能力和对垂直传播的保护作用外,新型CSFV E2与CD154亚单位疫苗对于发展中国家而言,是减毒活疫苗的一个有前景的替代选择。

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