Department of Molecular Biotechnology, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, B-9000 Ghent, Belgium.
Vaccine. 2010 Apr 19;28(18):3095-105. doi: 10.1016/j.vaccine.2010.02.064. Epub 2010 Mar 1.
We have demonstrated that vaccination of turkeys with an unformulated DNA vaccine induces significant protection against Chlamydophila (Cp.) psittaci infections. Nevertheless, the immunogenicity of the DNA vaccine can still be improved by increasing translation and transfection efficiency. Therefore, the ompA codon was adapted to the codon usage in birds, resulting in pcDNA1/MOMP(opt). To increase gene transfer, polyplexes of pcDNA1/MOMP(opt)-EGFP with different cationic polymers, such as linear and branched polyethyleneimine (lPEI and brPEI) and starburst PAMAM dendrimers, and lipoplexes with cationic DOTAP/DOPE liposomes were created. Transfection of lPEI and brPEI polyplexes with an N/P ratio of 8 resulted in the highest transfection efficiencies, but lPEI polyplexes were completely destroyed following nebulisation. Secondly, we examined the capacity of nebulised or intramuscularly (IM) administered brPEI-pcDNA1/MOMP(opt) to induce a significant protective immune response in SPF turkeys experimentally infected with 10(8) TCID(50) of a virulent Cp. psittaci strain. Results were compared to IM administration of naked plasmid DNA and to results of non-vaccinated animals. Intramuscular administration of brPEI-pcDNA1/MOMP(opt) increased the immunogenicity of the Cp. psittaci DNA vaccine as compared to IM administration of pcDNA1/MOMP(opt) or aerosol delivery of brPEI-pcDNA1/MOMP(opt). Improved immunogenicity was correlated with increased protection. Vaccinated groups were significantly protected against Cp. psittaci challenge.
我们已经证明,给火鸡接种未经配制的 DNA 疫苗可显著预防鹦鹉热衣原体(Cp.)感染。然而,通过提高翻译和转染效率,仍可以提高 DNA 疫苗的免疫原性。因此,ompA 密码子适应了鸟类的密码子用法,从而产生了 pcDNA1/MOMP(opt)。为了增加基因转移,用不同的阳离子聚合物,如线性和支化聚乙烯亚胺(lPEI 和 brPEI)和星型聚酰胺胺树枝状大分子,以及阳离子 DOTAP/DOPE 脂质体,制备了 pcDNA1/MOMP(opt)-EGFP 的多聚物和脂复合物。lPEI 和 brPEI 多聚物的 N/P 比为 8 时转染效率最高,但 lPEI 多聚物在雾化后完全被破坏。其次,我们研究了雾化或肌肉内(IM)给予 brPEI-pcDNA1/MOMP(opt)是否能够在 SPF 火鸡中诱导对强毒 Cp. psittaci 菌株 10(8)TCID(50)感染的显著保护免疫反应。结果与 IM 给予裸质粒 DNA 和非接种动物的结果进行了比较。与 IM 给予 pcDNA1/MOMP(opt)或雾化给予 brPEI-pcDNA1/MOMP(opt)相比,IM 给予 brPEI-pcDNA1/MOMP(opt)增加了 Cp. psittaci DNA 疫苗的免疫原性。免疫原性提高与保护增强相关。接种组对 Cp. psittaci 攻击有明显的保护作用。