Karpenko Larisa I, Apartsin Evgeny K, Dudko Sergei G, Starostina Ekaterina V, Kaplina Olga N, Antonets Denis V, Volosnikova Ekaterina A, Zaitsev Boris N, Bakulina Anastasiya Yu, Venyaminova Aliya G, Ilyichev Alexander A, Bazhan Sergei I
State Research Center of Virology and Biotechnology "Vector", Koltsovo, 630559 Novosibirsk Region, Russia.
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, 630090 Novosibirsk, Russia.
Vaccines (Basel). 2020 Dec 1;8(4):718. doi: 10.3390/vaccines8040718.
According to current data, an effective Ebola virus vaccine should induce both humoral and T-cell immunity. In this work, we focused our efforts on methods for delivering artificial T-cell immunogen in the form of a DNA vaccine, using generation 4 polyamidoamine dendrimers (PAMAM G4) and a polyglucin:spermidine conjugate (PG).
Optimal conditions were selected for obtaining complexes of previously developed DNA vaccines with cationic polymers. The sizes, mobility and surface charge of the complexes with PG and PAMAM 4G have been determined. The immunogenicity of the obtained vaccine constructs was investigated in BALB/c mice.
It was shown that packaging of DNA vaccine constructs both in the PG envelope and the PAMAM 4G envelope results in an increase in their immunogenicity as compared with the group of mice immunized with the of vector plasmid pcDNA3.1 (a negative control). The highest T-cell responses were shown in mice immunized with complexes of DNA vaccines with PG and these responses significantly exceeded those in the groups of animals immunized with both the combination of naked DNAs and the combination DNAs coated with PAMAM 4G. In the group of animals immunized with complexes of the DNA vaccines with PAMAM 4G, no statistical differences were found in the ability to induce T-cell responses, as compared with the group of mice immunized with the combination of naked DNAs.
The PG conjugate can be considered as a promising and safe means to deliver DNA-based vaccines. The use of PAMAM requires further optimization.
根据现有数据，一种有效的埃博拉病毒疫苗应能诱导体液免疫和T细胞免疫。在本研究中，我们致力于研究以DNA疫苗形式递送人工T细胞免疫原的方法，使用第4代聚酰胺-胺树枝状大分子（PAMAM G4）和聚葡萄糖-亚精胺缀合物（PG）。
选择最佳条件以获得先前开发的DNA疫苗与阳离子聚合物的复合物。已测定了与PG和PAMAM 4G形成的复合物的大小、迁移率和表面电荷。在BALB/c小鼠中研究了所得疫苗构建体的免疫原性。
结果表明，与用载体质粒pcDNA3.1免疫的小鼠组（阴性对照）相比，将DNA疫苗构建体包装在PG包膜和PAMAM 4G包膜中均会导致其免疫原性增加。在用DNA疫苗与PG的复合物免疫的小鼠中显示出最高的T细胞反应，这些反应显著超过了用裸DNA组合以及用PAMAM 4G包被的DNA组合免疫的动物组中的反应。在用DNA疫苗与PAMAM 4G的复合物免疫的动物组中，与用裸DNA组合免疫的小鼠组相比，在诱导T细胞反应的能力上未发现统计学差异。
PG缀合物可被视为递送基于DNA的疫苗的一种有前景且安全的手段。PAMAM的使用需要进一步优化。
