用于呼吸道病原体黏膜免疫的纳米佐剂干粉疫苗。

Nano-adjuvanted dry powder vaccine for the mucosal immunization against airways pathogens.

作者信息

Canelli Elena, Ferrari Luca, Borghetti Paolo, Candela Francesco, Abiakam Nkemjika Sopuru, Bianchera Annalisa, Buttini Francesca, Magi Gian Enrico, Sonvico Fabio, Martelli Paolo, Bettini Ruggero

机构信息

Department of Veterinary Science, University of Parma, Parma, Italy.

Advanced Drug Delivery Research Laboratory, Department of Food and Drug, University of Parma, Parma, Italy.

出版信息

Front Vet Sci. 2023 Mar 14;10:1116722. doi: 10.3389/fvets.2023.1116722. eCollection 2023.

Abstract

Nasal vaccination has been shown to provide optimal protection against respiratory pathogens. However, mucosal vaccination requires the implementation of specific immunization strategies to improve its effectiveness. Nanotechnology appears a key approach to improve the effectiveness of mucosal vaccines, since several nanomaterials provide mucoadhesion, enhance mucosal permeability, control antigen release and possess adjuvant properties. is the main causative agent of enzootic pneumonia in pigs, a respiratory disease responsible for considerable economic losses in the pig farming worldwide. The present work developed, characterized, and tested an innovative dry powder nasal vaccine, obtained from the deposition on a solid carrier of an inactivated antigen and a chitosan-coated nanoemulsion, as an adjuvant. The nanoemulsion was obtained through a low-energy emulsification technique, a method that allowed to achieve nano droplets in the order of 200 nm. The oil phase selected was alpha-tocopherol, sunflower oil, and poly(ethylene glycol) hydroxystearate used as non-ionic tensioactive. The aqueous phase contained chitosan, which provides a positive charge to the emulsion, conferring mucoadhesive properties and favoring interactions with inactivated . Finally, the nanoemulsion was layered with a mild and scalable process onto a suitable solid carrier (., lactose, mannitol, or calcium carbonate) to be transformed into a solid dosage form for administration as dry powder. In the experimental study, the nasal vaccine formulation with calcium carbonate was administered to piglets and compared to intramuscular administration of a commercial vaccine and of the dry powder without antigen, aimed at evaluating the ability of IN vaccination to elicit an local immune response and a systemic immune response. Intranasal vaccination was characterized by a significantly higher immune response in the nasal mucosa at 7 days post-vaccination, elicited comparable levels of -specific IFN-γ secreting cells and comparable, if not higher, responsiveness of B cells expressing IgA and IgG in peripheral blood mononuclear cells, with those detected upon a conventional intramuscular immunization. In conclusion, this study illustrates a simple and effective strategy for the development of a dry powder vaccine formulation for nasal administration which could be used as alternative to current parenteral commercial vaccines.

摘要

鼻腔接种已被证明能为抵御呼吸道病原体提供最佳保护。然而,黏膜接种需要实施特定的免疫策略来提高其有效性。纳米技术似乎是提高黏膜疫苗有效性的关键方法,因为几种纳米材料具有黏膜黏附性、能增强黏膜通透性、控制抗原释放并具备佐剂特性。猪肺炎支原体是猪地方流行性肺炎的主要病原体,这种呼吸道疾病在全球养猪业中造成了相当大的经济损失。本研究开发、表征并测试了一种创新的干粉鼻腔疫苗,该疫苗由灭活抗原沉积在固体载体上以及壳聚糖包被的纳米乳剂作为佐剂制成。纳米乳剂通过低能乳化技术获得,该方法可实现约200纳米大小的纳米液滴。所选油相为α-生育酚、葵花籽油和用作非离子表面活性剂的聚(乙二醇)羟基硬脂酸酯。水相含有壳聚糖,壳聚糖为乳剂提供正电荷,赋予其黏膜黏附特性并有利于与灭活猪肺炎支原体相互作用。最后,通过温和且可扩展的工艺将纳米乳剂分层到合适的固体载体(如乳糖、甘露醇或碳酸钙)上,转化为固体剂型以便作为干粉给药。在实验研究中,将含碳酸钙的鼻腔疫苗制剂给予仔猪,并与肌肉注射商业疫苗以及不含抗原的干粉进行比较,旨在评估鼻腔接种引发局部免疫反应和全身免疫反应的能力。鼻腔接种的特点是在接种后7天鼻腔黏膜中的免疫反应显著更高,在外周血单核细胞中引发的猪肺炎支原体特异性IFN-γ分泌细胞水平相当,表达IgA和IgG的B细胞反应性即使不更高也相当,与传统肌肉免疫时检测到的情况相当。总之,本研究阐明了一种开发用于鼻腔给药的干粉疫苗制剂的简单有效策略,该制剂可作为当前肠胃外商业疫苗的替代品。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/31fb/10043307/e7bea6a49f3f/fvets-10-1116722-g0001.jpg

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