Gorantala Jyotsna, Grover Sonam, Rahi Amit, Chaudhary Prerna, Rajwanshi Ravi, Sarin Neera Bhalla, Bhatnagar Rakesh
Laboratory of Molecular Biology and Genetic Engineering, School of Biotechnology, Jawaharlal Nehru University, New Delhi, India.
Molecular Technology Laboratory, School of Biotechnology, Jawaharlal Nehru University, New Delhi, India.
J Biotechnol. 2014 Apr 20;176:1-10. doi: 10.1016/j.jbiotec.2014.01.033. Epub 2014 Feb 15.
In concern with frequent recurrence of anthrax in endemic areas and inadvertent use of its spores as biological weapon, the development of an effective anthrax vaccine suitable for both human and veterinary needs is highly desirable. A simple oral delivery through expression in plant system could offer promising alternative to the current methods that rely on injectable vaccines extracted from bacterial sources. In the present study, we have expressed protective antigen (PA) gene in Indian mustard by Agrobacterium-mediated transformation and in tobacco by plastid transformation. Putative transgenic lines were verified for the presence of transgene and its expression by molecular analysis. PA expressed in transgenic lines was biologically active as evidenced by macrophage lysis assay. Intraperitoneal (i.p.) and oral immunization with plant PA in murine model indicated high serum PA specific IgG and IgA antibody titers. PA specific mucosal immune response was noted in orally immunized groups. Further, antibodies indicated lethal toxin neutralizing potential in-vitro and conferred protection against in-vivo toxin challenge. Oral immunization experiments demonstrated generation of immunoprotective response in mice. Thus, our study examines the feasibility of oral PA vaccine expressed in an edible plant system against anthrax.
鉴于炭疽病在流行地区频繁复发以及其孢子被无意用作生物武器,开发一种适用于人类和兽医需求的有效炭疽疫苗非常必要。通过在植物系统中表达进行简单口服给药,可能为目前依赖从细菌来源提取的注射用疫苗的方法提供有前景的替代方案。在本研究中,我们通过农杆菌介导的转化在印度芥菜中表达了保护性抗原(PA)基因,并通过质体转化在烟草中表达了该基因。通过分子分析对推定的转基因株系进行了转基因存在及其表达的验证。巨噬细胞裂解试验证明,转基因株系中表达的PA具有生物活性。在小鼠模型中,用植物PA进行腹腔注射(i.p.)和口服免疫表明血清中PA特异性IgG和IgA抗体滴度很高。口服免疫组出现了PA特异性黏膜免疫反应。此外,抗体在体外显示出致死毒素中和潜力,并在体内毒素攻击中提供保护。口服免疫实验证明在小鼠中产生了免疫保护反应。因此,我们的研究检验了在可食用植物系统中表达的口服PA疫苗预防炭疽病的可行性。