College of Marine Life Science, Ocean University of China, Qingdao 266003, PR China.
College of Marine Life Science, Ocean University of China, Qingdao 266003, PR China.
Int J Biol Macromol. 2016 Oct;91:716-23. doi: 10.1016/j.ijbiomac.2016.06.015. Epub 2016 Jun 7.
This study aimed to investigate the efficacy of nanoparticles based on chitosan as a vehicle for oral antigen delivery in fish vaccination. Carboxymethyl chitosan/chitosan nanoparticles (CMCS/CS-NPs) loaded extracellular products (ECPs) of Vibrio anguillarum were successfully developed by ionic gelation method. The prepared ECPs-loaded CMCS/CS-NPs were characterized for various parameters including morphology, particle size (312±7.18nm), zeta potential (+17.4±0.38mV), loading efficiency (57.8±2.54%) and stability under the simulated gastrointestinal (GI) tract conditions in turbot. The in vitro profile showed that the cumulative release of ECPs from nanoparticles was higher in pH 7.4 (58%) than in pH 2.0 (37%) and pH 4.5 (29%) after 48h. Fluorescein isothiocyanate-labeled bovine serum albumin (FITC-BSA) was used as model protein antigen and encapsulated in CMCS/CS-NPs for investigating the biodistribution of antigen after oral delivery to turbot in 24h. Oral immunization of ECPs-loaded CMCS/CS-NPs group in turbot showed elevated specific antibody and higher concentrations of lysozyme activity and complement activity in fish serum than ECPs solution. CMCS/CS-NPs loaded with ECPs could enhance both adaptive and innate immune responses than the group treated with ECPs solution and suggested to be a potential antigen delivery system.
本研究旨在探讨壳聚糖基纳米粒子作为鱼类疫苗口服抗原传递载体的功效。采用离子凝胶法成功制备了载有鳗弧菌胞外产物（ECPs）的羧甲基壳聚糖/壳聚糖纳米粒子（CMCS/CS-NPs）。对载有 ECPs 的 CMCS/CS-NPs 的各种参数进行了表征，包括形态、粒径（312±7.18nm）、Zeta 电位（+17.4±0.38mV）、载药效率（57.8±2.54%）和在大菱鲆模拟胃肠道（GI）条件下的稳定性。体外研究结果表明，在 pH7.4 时，纳米粒子中 ECPs 的累积释放率（58%）高于 pH2.0（37%）和 pH4.5（29%），48h 后释放率达到 58%。荧光素异硫氰酸酯标记牛血清白蛋白（FITC-BSA）作为模型蛋白抗原，包封在 CMCS/CS-NPs 中，用于研究 24h 后口服给予大菱鲆时抗原的体内分布情况。大菱鲆口服 ECPs 负载的 CMCS/CS-NPs 组显示，与 ECPs 溶液组相比，鱼血清中的特异性抗体水平升高，溶菌酶活性和补体活性更高。载有 ECPs 的 CMCS/CS-NPs 可增强适应性和固有免疫反应，优于 ECPs 溶液组，提示其可能成为一种潜在的抗原传递系统。
