Chia Min-Yuan, Hsiao Shih-Hsuan, Chan Hui-Ting, Do Yi-Yin, Huang Pung-Ling, Chang Hui-Wen, Tsai Yi-Chieh, Lin Chun-Ming, Pang Victor Fei, Jeng Chian-Ren
Graduate Institute of Veterinary Medicine, School of Veterinary Medicine, Taipei 106, Taiwan, ROC.
Vet Immunol Immunopathol. 2010 Jun 15;135(3-4):234-42. doi: 10.1016/j.vetimm.2009.12.003. Epub 2009 Dec 22.
The aim of the study was to evaluate the immunogenicity of the ORF5-encoded major envelop glycoprotein 5 (GP5) of porcine reproductive and respiratory syndrome virus (PRRSV) expressed in tobacco plant as a potential pig oral vaccine in protection against PRRSV infection. Six-week-old PRRSV-free pigs were fed four times orally with 50g of chopped fresh GP5 transgenic tobacco leaves (GP5-T) (GP5 reaching 0.011% of total soluble protein) or wild-type tobacco leaves (W-T) each on days 0, 14, 28, and 42. Samples of serum, saliva, and peripheral blood mononuclear cells (PBMCs) were collected on days -1, 6, 13, 20, 27, 34, 41, and 48 after the initial oral vaccination. A similar vaccination-dependent gradual increase in the responses of serum and saliva anti-PRRSV total IgG and IgA, respectively, and in the levels of PRRSV-specific blastogenic response of PBMCs was seen in GP5-T-treated pigs; all statistically significant elevations occurred after the 2nd vaccination and were revealed after 20 days post-initial oral vaccination (DPIOV). Pigs fed on GP5-T also developed serum neutralizing antibodies to PRRSV at a titer of 1:4-1:8 after the 4th vaccination by 48 DPIOV. No detectable anti-PRRSV antibody responses and PRRSV-specific blastogenic response were seen in W-T-treated pigs. The present study has demonstrated that pigs fed on GP5-T could develop specific mucosal as well as systemic humoral and cellular immune responses against PRRSV. The results also support that transgenic plant as GP5-T can be an effective system for oral delivery of recombinant subunit vaccines in pigs.
本研究旨在评估在烟草植物中表达的猪繁殖与呼吸综合征病毒(PRRSV)的开放阅读框5(ORF5)编码的主要包膜糖蛋白5(GP5)作为一种潜在的猪口服疫苗预防PRRSV感染的免疫原性。六周龄无PRRSV的仔猪在第0、14、28和42天分别四次口服50克切碎的新鲜GP5转基因烟草叶(GP5-T)(GP5占总可溶性蛋白的0.011%)或野生型烟草叶(W-T)。在初次口服疫苗接种后的第-1、6、13、20、27、34、41和48天收集血清、唾液和外周血单核细胞(PBMC)样本。在GP5-T处理的猪中,分别观察到血清和唾液中抗PRRSV总IgG和IgA反应以及PBMC中PRRSV特异性增殖反应的类似疫苗接种依赖性逐渐增加;所有统计学上显著的升高均发生在第二次疫苗接种后,并在初次口服疫苗接种后20天(DPIOV)显现。在第4次疫苗接种后至48 DPIOV时,以GP5-T喂养的猪也产生了滴度为1:4 - 1:8的针对PRRSV的血清中和抗体。在W-T处理的猪中未观察到可检测到的抗PRRSV抗体反应和PRRSV特异性增殖反应。本研究表明,以GP5-T喂养的猪可产生针对PRRSV的特异性黏膜以及全身性体液和细胞免疫反应。结果还支持,作为GP5-T的转基因植物可成为猪口服重组亚单位疫苗的有效递送系统。