Dendritic cell targeted chitosan nanoparticles for nasal DNA immunization against SARS CoV nucleocapsid protein.
Dendritic cell targeted chitosan nanoparticles for nasal DNA immunization against SARS CoV nucleocapsid protein.
复制标题
DOI：
10.1021/mp200553x
复制
发表时间：
2012-04-02
影响因子：
4.9
通讯作者：
Suresh MR
中科院分区：
文献类型：
--
作者：
Raghuwanshi D;Mishra V;Das D;Kaur K;Suresh MR
本文研究了树突状细胞（DC）靶向质粒DNA负载生物素化壳聚糖纳米颗粒鼻腔免疫非典型肺炎冠状病毒（SARS-CoV）核衣壳(N)蛋白作为抗原的制备方法和体内效果。在病毒进入部位诱导抗原特异性粘膜和全身免疫反应是疫苗设计的主要挑战。在这里，我们设计了一种非侵入性受体介导的基因递送策略。采用复杂的凝聚工艺制备了负载pDNA的生物素化壳聚糖纳米颗粒，并对其大小、形状、表面电荷、质粒负载和抗核酸酶酶切性进行了表征。利用双功能融合蛋白（bfFp）载体靶向负载pDNA的生物素化壳聚糖纳米颗粒，实现DC选择性靶向。bfFp是一种重组融合蛋白，由截断的核心链亲和素与抗dec -205单链抗体（scFv）融合而成。融合蛋白的核心链亲和素臂与生物素化纳米颗粒结合，而抗DEC-205 scFv则赋予DC DEC-205受体靶向特异性。我们证明鼻内给药bfFp靶向制剂以及抗cd40 DC成熟刺激增强了黏膜IgA和全身针对N蛋白的IgG的大小。该策略导致检测到N蛋白特异性全身IgG和鼻腔IgA抗体水平的增强。然而，在鼻内递送裸pDNA后，未检测到粘膜和全身免疫反应。对肌内和鼻内途径靶向制剂的平行比较表明，与鼻内途径相比，肌内途径在诱导全身IgG反应方面优于鼻内途径。我们的研究结果表明，通过非侵入性鼻内途径靶向pDNA递送可以作为设计低剂量疫苗的一种策略。
This work investigates the formulation and in vivo efficacy of dendritic cell (DC) targeted plasmid DNA loaded biotinylated chitosan nanoparticles for nasal immunization against nucleocapsid (N) protein of severe acute respiratory syndrome coronavirus (SARS-CoV) as antigen. The induction of antigen-specific mucosal and systemic immune response at the site of virus entry is a major challenge for vaccine design. Here, we designed a strategy for non-invasive receptor mediated gene delivery to nasal resident DCs. The pDNA loaded biotinylated chitosan nanoparticles were prepared using a complex coacervation process and characterized for size, shape, surface charge, plasmid loading and protection against nuclease digestion. The pDNA loaded biotinylated chitosan nanoparticles were targeted with bifunctional fusion protein (bfFp) vector for achieving DC selective targeting. The bfFp is a recombinant fusion protein consisting of truncated core-streptavidin fused with anti-DEC-205 single chain antibody (scFv). The core-streptavidin arm of fusion protein binds with biotinylated nanoparticles, while anti-DEC-205 scFv imparts targeting specificity to DC DEC-205 receptor. We demonstrate that intranasal administration of bfFp targeted formulations along with anti-CD40 DC maturation stimuli enhanced magnitude of mucosal IgA as well as systemic IgG against N protein. The strategy led to the detection of augmented levels of N protein specific systemic IgG and nasal IgA antibodies. However, following intranasal delivery of naked pDNA no mucosal and systemic immune responses were detected. A parallel comparison of targeted formulations using intramuscular and intranasal route showed that the intramuscular route is superior for induction of systemic IgG responses compared with the intranasal route. Our results suggest that targeted pDNA delivery through non-invasive intranasal route can be a strategy for designing low-dose vaccines.
登录
查看更多内容
DOI：
10.1083/jcb.151.3.673
发表时间：
2000-10-30
期刊：
The Journal of cell biology
影响因子：
--
作者：
Mahnke K;Guo M;Lee S;Sepulveda H;Swain SL;Nussenzweig M;Steinman RM
通讯作者：
Steinman RM
影响因子：
15.3
作者：
Bonifaz, L;Bonnyay, D;Mahnke, K;Rivera, M;Nussenzweig, MC;Steinman, RM
通讯作者：
Steinman, RM
影响因子：
14
作者：
Corsi, K;Chellat, F;Fernandes, JC
通讯作者：
Fernandes, JC
影响因子：
5.8
作者：
Khatri, Kapil;Goyal, Amit K.;Vyas, Suresh P.
通讯作者：
Vyas, Suresh P.
DOI：
10.1038/nrg2432
发表时间：
2008-10
期刊：
Nature reviews. Genetics
影响因子：
--
作者：
通讯作者：
--