Jesus Sandra, Soares Edna, Borges Olga
Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504, Coimbra, Portugal.
Faculty of Pharmacy, University of Coimbra, 3000-548, Coimbra, Portugal.
Methods Mol Biol. 2016;1404:697-713. doi: 10.1007/978-1-4939-3389-1_45.
Several evidences converge on the idea that among the mucosal administration routes, the nasal mucosa is the most attractive site for the delivery of vaccines. Mucoadhesive particulate adjuvants should be able to increase the residence time of antigens in nasal cavity in order to increase their probability of being taken up by nasopharynx-associated lymphoid tissue (NALT) cells and subsequently to initiate the innate and adaptive immune response. Focusing on chitosan, a mucoadhesive biopolymer, we describe in this chapter a method to prepare antigen loaded chitosan nanoparticles and a second method to prepare antigen loaded poly-ε-caprolactone/chitosan nanoparticles. Additionally the methodology for the assessment of mucoadhesivity of the delivery system is also described. The two critical procedures in mice intranasal immunization experiments include challenges in the intranasal administration itself due to the small mouse nose, and the other is related with the collection of mucosal secretions to assess the sIgA. The techniques are difficult to perform without advanced training. Therefore, protocols followed in our laboratory, as well as some tips, are described in this chapter.
在黏膜给药途径中,鼻腔黏膜是疫苗递送最具吸引力的部位。黏膜黏附性微粒佐剂应能够增加抗原在鼻腔中的停留时间,以提高其被鼻咽相关淋巴组织(NALT)细胞摄取的概率,进而启动先天性和适应性免疫反应。本章聚焦于一种黏膜黏附性生物聚合物壳聚糖,描述了一种制备负载抗原的壳聚糖纳米颗粒的方法以及另一种制备负载抗原的聚ε-己内酯/壳聚糖纳米颗粒的方法。此外,还介绍了评估递送系统黏膜黏附性的方法。小鼠鼻内免疫实验中的两个关键步骤,一是由于小鼠鼻子较小,鼻内给药本身具有挑战性,另一个是与收集黏膜分泌物以评估分泌型免疫球蛋白A(sIgA)有关。如果没有经过高级培训,这些技术很难实施。因此,本章将介绍我们实验室遵循的方案以及一些技巧。