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酵母表达的经典猪瘟病毒糖蛋白 E2 诱导保护性免疫应答。

Yeast-expressed classical swine fever virus glycoprotein E2 induces a protective immune response.

机构信息

Graduate Institute of Veterinary Microbiology, College of Veterinary Medicine, National Chung Hsing University, 250 Kuo Kuang Road, Taichung 40227, Taiwan, ROC.

出版信息

Vet Microbiol. 2009 Nov 18;139(3-4):369-74. doi: 10.1016/j.vetmic.2009.06.027. Epub 2009 Jun 21.

Abstract

Classical swine fever (CSF) is an economically important swine disease worldwide. The glycoprotein E2 of classical swine fever virus (CSFV) is a viral antigen that can induce a protective immune response against CSF. A recombinant E2 protein was constructed using the yeast Pichia pastoris expression system and evaluated for its vaccine efficacy. The yeast-expressed E2 (yE2) was shown to have N-linked glycosylation and to form homodimer molecules. Four 6-week-old specified-pathogen-free (SPF) piglets were intramuscularly immunized with yE2 twice at 3-week intervals. All yE2-vaccinated pigs could mount an anamnestic response after booster vaccination with neutralizing antibody titers ranging from 1:96 to 1:768. Neutralizing antibody titers at 10 weeks post booster vaccination ranged from 1:16 to 1:64. At this time, the pigs were subjected to challenge infection with a dose of 1x10(5)TCID(50) (50% tissue culture infective dose) virulent CSFV strain. At 1 week post challenge infection, all of the yE2-immunized pigs were alive and without symptoms or signs of CSF. Neutralizing antibody titers at this time ranged from 1:4,800 to 1:12,800 and even to 1:51,200 one week later. In contrast, the control pigs continuously exhibited signs of CSF and had to be euthanized because of severe clinical symptoms at 6 days post challenge infection. All of the yE2-vaccinated pigs were E(rns) antibody negative and had seroconverted against E(rns) by post challenge day 11, suggesting that yE2 is a potential DIVA (differentiating infected from vaccinated animals) vaccine. The yeast-expressed E2 protein retains correct immunogenicity and is able to induce a protective immune response against CSFV infection.

摘要

古典猪瘟(CSF)是一种在全球范围内对养猪业具有重要经济意义的猪病。猪瘟病毒(CSFV)的糖蛋白 E2 是一种病毒抗原,可诱导针对 CSF 的保护性免疫反应。本研究使用毕赤酵母表达系统构建了重组 E2 蛋白,并评估了其疫苗效力。结果表明,酵母表达的 E2(yE2)具有 N 连接糖基化,并形成同源二聚体分子。4 头 6 周龄无特定病原体(SPF)仔猪间隔 3 周肌内免疫 yE2 两次。所有 yE2 疫苗接种的猪在加强免疫后都能产生回忆应答,中和抗体滴度范围为 1:96 至 1:768。加强免疫后 10 周的中和抗体滴度范围为 1:16 至 1:64。此时,这些猪用 1x10(5)TCID(50)(50%组织培养感染剂量)强毒猪瘟病毒株进行攻毒感染。攻毒感染后 1 周,所有 yE2 免疫的猪均存活且无 CSF 症状或体征。此时的中和抗体滴度范围为 1:4,800 至 1:12,800,甚至 1 周后为 1:51,200。相比之下,对照猪在攻毒感染后 6 天持续出现 CSF 症状和体征,不得不因严重的临床症状而被安乐死。所有 yE2 疫苗接种的猪均为 E(rns)抗体阴性,且在攻毒后第 11 天转为 E(rns)抗体阳性,表明 yE2 是一种潜在的区分感染动物和免疫动物的(DIVA)疫苗。酵母表达的 E2 蛋白保留了正确的免疫原性,并能诱导针对 CSFV 感染的保护性免疫反应。

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