Calegari Luan P, Dias Roberto S, de Oliveira Michelle D, Pessoa Carine Ribeiro, de Oliveira André S, Oliveira Ana F C S, da Silva Cynthia C, Fonseca Flavio G, Versiani Alice F, De Paula Sérgio O
Federal University of Viçosa, Avenida Peter Henry Rolfs, s/n, Campus Universitário, Viçosa, MG, 36570-900, Brazil.
Federal Institute of Education, Science and Technology of North of Minas Gerais, R. Gabriel Passos, 259, Centro, Montes Claros, MG, 39400-112, Brazil.
J Nanobiotechnology. 2016 Jul 27;14(1):61. doi: 10.1186/s12951-016-0196-7.
In recent times, studies have demonstrated that carbon nanotubes are good candidates for use as vehicles for transfection of exogenous material into the cells. However, there are few studies evaluating the behavior of carbon nanotubes as DNA vectors and few of these studies have used multi-walled carbon nanotubes (MWCNTs) or carboxylated MWCNTs. Thus, this study aims to assess the MWCNTs' (carboxylated or not) efficiency in the increase in expression of the tetravalent vaccine candidate (TVC) plasmid vector for dengue virus in vitro using Vero cells, and in vivo, through the intramuscular route, to evaluate the immunological response profile.
Multi-walled carbon nanotubes internalized by Vero cells, have been found in the cytoplasm and nucleus associated with the plasmid. However, it was not efficient to increase the messenger ribonucleic acid (mRNA) compared to the pure vaccine candidate associated with Lipofectamine(®) 2000. The in vivo experiments showed that the use of intramuscular injection of the TVC in combination with MWCNTs reduced the immune response compared to pure TVC, in a general way, although an increase was observed in the population of the antibody-producing B cells, as compared to pure TVC.
The results confirm the data found by other authors, which demonstrate the ability of nanotubes to penetrate target cells and reach both the cytoplasm and the cell nucleus. The cytotoxicity values are also in accordance with the literature, which range from 5 to 20 µg/mL. This has been found to be 10 µg/mL in this study. Although the expression levels are higher in cells that receive the pure TVC transfected using Lipofectamine(®) 2000, the nanotubes show an increase in B-cells producing antibodies.
近年来,研究表明碳纳米管是将外源物质转染到细胞中的良好载体。然而,评估碳纳米管作为DNA载体行为的研究较少,且这些研究中很少使用多壁碳纳米管(MWCNT)或羧化MWCNT。因此,本研究旨在评估MWCNT(羧化或未羧化)在体外使用Vero细胞增加登革病毒四价疫苗候选物(TVC)质粒载体表达的效率,以及在体内通过肌肉注射途径评估免疫反应谱。
Vero细胞内化的多壁碳纳米管已在与质粒相关的细胞质和细胞核中被发现。然而,与与Lipofectamine® 2000相关的纯疫苗候选物相比,它在增加信使核糖核酸(mRNA)方面效率不高。体内实验表明,与纯TVC相比,肌肉注射TVC与MWCNT联合使用总体上降低了免疫反应,尽管与纯TVC相比,产生抗体的B细胞群体有所增加。
结果证实了其他作者发现的数据,这些数据表明纳米管能够穿透靶细胞并到达细胞质和细胞核。细胞毒性值也与文献一致,范围为5至20μg/mL。本研究中发现为10μg/mL。尽管在接受使用Lipofectamine® 2000转染的纯TVC的细胞中表达水平较高,但纳米管显示产生抗体的B细胞有所增加。