Bilayer films for mucosal (genetic) immunization via the buccal route in rabbits
Bilayer films for mucosal (genetic) immunization via the buccal route in rabbits
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DOI：
10.1023/a:1016454003450
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发表时间：
2002-07-01
影响因子：
3.7
通讯作者：
Mumper, RJ
中科院分区：
文献类型：
--
作者：
Cui, ZR;Mumper, RJ
目的.口腔颊粘膜可能是粘膜免疫的理想部位，允许无针施用具有成本效益的疫苗。开发了一种新的粘膜粘附双层膜，以测试这种免疫途径在家兔中的可行性。使用不同比例的Noveon和Eudragit S-100作为粘膜粘附层和药用蜡作为不可渗透的背衬层开发双层膜。最佳的3/8英寸胶片后加载100 μ g质粒DNA（CMV-β-gal）或β-半乳糖苷酶蛋白。测定了后载抗原的体外释放速率和稳定性。于第0、7和14天将膜贴敷于家兔的颊囊，测定至第28天对β-gal的体液和脾细胞增殖免疫应答，并与常规皮下注射佐剂蛋白后的应答进行比较。Noveon和Eudragit S-100的重量比对双层膜的粘附时间有显著影响。后加载的质粒DNA和β-gal在从双层膜释放后保持稳定（两者在2小时内的释放类似于60 -80%）。使用含有质粒DNA的新型双层膜（109 +/-6-μ m厚度）的颊部免疫导致与皮下蛋白质注射相当的抗原特异性IgG滴度。质粒DNA经口腔途径免疫家兔后，均出现脾细胞增殖免疫反应，而蛋白抗原经皮下途径免疫家兔后，均未出现脾细胞增殖免疫反应。证明了这些新型双层膜用于口腔（遗传）免疫的可行性。
Purpose. The oral buccal mucosa may be an ideal site for mucosal immunization, allowing for the needle-free administration of cost-effective vaccines. A novel mucoadhesive bilayer film was developed to test the feasibility of this route of immunization in rabbits.Methods. Bilayer films were developed using different ratios of Noveon and Eudragit S-100 as the mucoadhesive layer and a pharmaceutical wax as the impermeable backing layer. Optimal 3/8-inch films were post-loaded with 100 mug of plasmid DNA (CMV-beta-gal) or beta-galactosidase protein. The in vitro release rates and stability of the postloaded antigens were determined. The films were applied to the buccal pouch of rabbits on days 0, 7, and 14, and the humoral and splenocyte proliferative immune responses to beta-gal were determined through day 28 and compared to those responses after conventional subcutaneous injection of adjuvanted protein.Results. The weight ratio of Noveon and Eudragit S-100 had a significant effect on adhesion time of bilayer films. Postloaded plasmid DNA and beta-gal remained stable after being released from bilayer films (release of similar to60-80% in 2 h for both). Buccal immunization using novel bilayer films (109 +/- 6-mum thickness) containing plasmid DNA led to comparable antigen-specific IgG titer to that of subcutaneous protein injection. All rabbits immunized with plasmid DNA via the buccal route but none by the subcutaneous route with protein antigen demonstrated splenocyte proliferative immune responses.Conclusion. The feasibility of buccal (genetic) immunization with these novel bilayer films was demonstrated.