Recombinant plants provide a new approach to the production of bacterial polysaccharide for vaccines.
Recombinant plants provide a new approach to the production of bacterial polysaccharide for vaccines.
复制标题
DOI：
10.1371/journal.pone.0088144
复制
发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Andrew PW
中科院分区：
文献类型：
--
作者：
Smith CM;Fry SC;Gough KC;Patel AJ;Glenn S;Goldrick M;Roberts IS;Whitelam GC;Andrew PW
细菌多糖具有多种临床或工业用途。重组植物为大规模生产细菌多糖提供了可能，而且不会污染细菌毒素和抗原。我们通过利用pCambia2301载体和农杆菌介导的基因转移，在烟草中克隆和表达肺炎链球菌3型合成酶(Cps3S)基因，研究了这一方案的可行性。在植物中，重组合成酶聚合植物来源的UDP-葡萄糖和UDP-葡萄糖醛酸形成3型多糖。RT-PCR检测cps3S基因的表达，双向免疫扩散、Western blotting和高压纸片电泳法检测烟叶提取物中肺炎链球菌多糖的产生。因为它是抗肺炎球菌疫苗的一种成分，所以对植物来源的3型多糖的免疫原性进行了测试。在肺炎模型中，用重组植物提取物免疫的小鼠免受致死剂量肺炎球菌的攻击，免疫小鼠的血清抗肺炎球菌多糖抗体水平显著升高。这项研究证明了细菌多糖可以在植物中成功合成的原理，以及这些重组多糖可以用作疫苗来预防危及生命的感染。
Bacterial polysaccharides have numerous clinical or industrial uses. Recombinant plants could offer the possibility of producing bacterial polysaccharides on a large scale and free of contaminating bacterial toxins and antigens. We investigated the feasibility of this proposal by cloning and expressing the gene for the type 3 synthase (cps3S) of Streptococcus pneumoniae in Nicotinia tabacum, using the pCambia2301 vector and Agrobacterium tumefaciens-mediated gene transfer. In planta the recombinant synthase polymerised plant-derived UDP-glucose and UDP-glucuronic acid to form type 3 polysaccharide. Expression of the cps3S gene was detected by RT-PCR and production of the pneumococcal polysaccharide was detected in tobacco leaf extracts by double immunodiffusion, Western blotting and high-voltage paper electrophoresis. Because it is used a component of anti-pneumococcal vaccines, the immunogenicity of the plant-derived type 3 polysaccharide was tested. Mice immunised with extracts from recombinant plants were protected from challenge with a lethal dose of pneumococci in a model of pneumonia and the immunised mice had significantly elevated levels of serum anti-pneumococcal polysaccharide antibodies. This study provides the proof of the principle that bacterial polysaccharide can be successfully synthesised in plants and that these recombinant polysaccharides could be used as vaccines to protect against life-threatening infections.
登录
查看更多内容
影响因子：
168.9
作者：
Levine, MM;Levine, OS
通讯作者：
Levine, OS
影响因子：
17.3
作者：
MASON, HS;ARNTZEN, CJ
通讯作者：
ARNTZEN, CJ
影响因子：
3.2
作者：
Arrecubieta, C;Garcia, E;Lopez, R
通讯作者：
Lopez, R
影响因子：
6.2
作者：
Rigano, MM;Alvarez, ML;Walmsley, AM
通讯作者：
Walmsley, AM
影响因子：
6.4
作者：
DOUGLAS, RM;PATON, JC;HANSMAN, DJ
通讯作者：
HANSMAN, DJ