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作为一种具有成本效益的疫苗生产替代方案,在大肠杆菌中表达的嵌合木瓜环斑病毒样颗粒密集携带的猪圆环病毒2型保护性表位。

Porcine circovirus type 2 protective epitope densely carried by chimeric papaya ringspot virus-like particles expressed in Escherichia coli as a cost-effective vaccine manufacture alternative.

作者信息

Aguilera Brenda Eugenia, Chávez-Calvillo Gabriela, Elizondo-Quiroga Darwin, Jimenez-García Mónica Noemí, Carrillo-Tripp Mauricio, Silva-Rosales Laura, Hernández-Gutiérrez Rodolfo, Gutiérrez-Ortega Abel

机构信息

Unidad de Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco A.C., Normalistas 800, Colinas de la Normal, Guadalajara, Jalisco 44270, México.

Departamento de Ingeniería Genética, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, km 9.6 Libramiento Norte, Carretera Irapuato-León, Irapuato, Guanajuato 36821, México.

出版信息

Biotechnol Appl Biochem. 2017 May;64(3):406-414. doi: 10.1002/bab.1491. Epub 2016 Apr 20.

Abstract

Porcine circovirus type 2 (PCV2) still represents a major problem to the swine industry worldwide, causing high mortality rates in infected animals. Virus-like particles (VLPs) have gained attention for vaccine development, serving both as scaffolds for epitope expression and immune response enhancers. The commercial subunit vaccines against PCV2 consist of VLPs formed by the self-assembly of PCV2 capsid protein (CP) expressed in the baculovirus vector system. In this work, a PCV2 protective epitope was inserted into three different regions of papaya ringspot virus (PRSV) CP, namely, the N- and C-termini and a predicted antigenic region located near the N-terminus. Wild-type and chimeric CPs were modeled in silico, expressed in Escherichia coli, purified, and visualized by transmission electron microscopy. This is the first report that shows the formation of chimeric VLPs using PRSV as epitope-presentation scaffold. Moreover, it was found that PCV2 epitope localization strongly influences VLP length. Also, the estimated yields of the chimeric VLPs at a small-scale level ranged between 65 and 80 mg/L of culture medium. Finally, the three chimeric VLPs induced high levels of immunoglobulin G against the PCV2 epitope in immunized BALB/c mice, suggesting that these chimeric VLPs can be used for swine immunoprophylaxis against PCV2.

摘要

猪圆环病毒2型(PCV2)仍然是全球养猪业面临的一个主要问题,可导致感染动物的高死亡率。病毒样颗粒(VLP)在疫苗开发中受到关注,既作为表位表达的支架,也作为免疫反应增强剂。针对PCV2的商业亚单位疫苗由在杆状病毒载体系统中表达的PCV2衣壳蛋白(CP)自组装形成的VLP组成。在这项工作中,将一个PCV2保护性表位插入番木瓜环斑病毒(PRSV)CP的三个不同区域,即N端和C端以及位于N端附近的一个预测抗原区域。对野生型和嵌合CP进行了计算机模拟建模,在大肠杆菌中表达、纯化,并通过透射电子显微镜观察。这是第一份显示使用PRSV作为表位呈递支架形成嵌合VLP的报告。此外,发现PCV2表位定位强烈影响VLP长度。此外,小规模培养时嵌合VLP的估计产量在65至80 mg/L培养基之间。最后,三种嵌合VLP在免疫的BALB/c小鼠中诱导产生了高水平的针对PCV2表位的免疫球蛋白G,表明这些嵌合VLP可用于猪针对PCV2的免疫预防。

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