Sánchez O, Barrera M, Farnós O, Parra N C, Salgado E R, Saavedra P A, Meza C D, Rivas C I, Cortez-San Martín M, Toledo J R
Department of Pharmacology, School of Biological Sciences, Universidad de Concepción, Concepción, Chile.
Biotechnology and Biopharmaceuticals Laboratory, Department of Pathophysiology, School of Biological Sciences, Universidad de Concepción, Concepción, Chile Prometeo Project, Secretaría de Educación Superior, Quito, Ecuador.
Clin Vaccine Immunol. 2014 Dec;21(12):1628-34. doi: 10.1128/CVI.00416-14. Epub 2014 Oct 1.
Subunit recombinant vaccines against classical swine fever virus (CSFV) are a promising alternative to overcome practical and biosafety issues with inactivated vaccines. One of the strategies in evaluation under field conditions is the use of a new marker E2-based vaccine produced in the milk of adenovirally transduced goats. Previously we had demonstrated the efficacy of this antigen, which conferred early protection and long-lasting immunity in swine against CSFV infection. Here, we have used a simpler downstream process to obtain and formulate the recombinant E2 glycoprotein expressed in the mammary gland. The expression levels reached approximately 1.7 mg/ml, and instead of chromatographic separation of the antigen, we utilized a clarification process that eliminates the fat content, retains a minor amount of caseins, and includes an adenoviral inactivation step that improves the biosafety of the final formulation. In a vaccination and challenge experiment in swine, different doses of the E2 antigen contained within the clarified whey generated an effective immune response of neutralizing antibodies that protected all of the animals against a lethal challenge with CSFV. During the immunization and after challenge, the swine were monitored for adverse reactions related to the vaccine or symptoms of CSF, respectively. No adverse reactions or clinical signs of disease were observed in vaccinated animals, in which no replication of CSFV could be detected after challenge. Overall, we consider that the simplicity of the procedures proposed here is a further step toward the introduction and implementation of a commercial subunit vaccine against CSF.
针对经典猪瘟病毒(CSFV)的亚单位重组疫苗是克服灭活疫苗实际应用和生物安全问题的一种有前景的替代方案。在田间条件下评估的策略之一是使用一种基于新型标记E2的疫苗,该疫苗由腺病毒转导的山羊乳汁生产。此前我们已证明这种抗原的效力,它能为猪提供针对CSFV感染的早期保护和持久免疫力。在此,我们采用了一种更简单的下游工艺来获取和配制在乳腺中表达的重组E2糖蛋白。表达水平达到约1.7毫克/毫升,并且我们没有采用抗原的色谱分离方法,而是利用了一种澄清工艺,该工艺可去除脂肪含量,保留少量酪蛋白,并包括一个腺病毒灭活步骤,从而提高最终制剂的生物安全性。在猪的疫苗接种和攻毒实验中,澄清乳清中含有的不同剂量E2抗原产生了有效的中和抗体免疫反应,保护所有动物免受CSFV的致死性攻毒。在免疫期间和攻毒后,分别监测猪与疫苗相关的不良反应或CSF症状。在接种疫苗的动物中未观察到不良反应或疾病临床症状,攻毒后也未检测到CSFV的复制。总体而言,我们认为这里提出的程序简单性是朝着引入和实施针对CSF的商业亚单位疫苗迈出的又一步。