CTB AS AN ORAL ADJUVANT FOR PARAINFLUENZA VACCINE

这项提议的长期目标是开发一种商业 副流感病毒三价糖蛋白亚单位疫苗。因为 制备这些糖蛋白的高昂成本，这是一种实现 增强免疫反应是一个重要的目标，也是 这项提案的直接焦点。已经证明，霍乱毒素 B亚单位(CTB)不仅是一种有效的粘膜免疫原，而且 共同接种的对无关抗原的抗体反应的佐剂。这个 CTB对PI3糖蛋白亚单位疫苗的佐剂作用 已评估。两种糖蛋白，融合蛋白(F)和血凝素神经氨酸酶 将从病毒粒子或病毒中提纯PI3病毒的(Hn)蛋白 特异性单抗偶联亲和柱法感染细胞 抗F或HN抗体。糖蛋白将通过以下方式与CTB偶联 不同的方法和结合物各组分的抗原性 由于共轭CTB与GM1神经节苷脂的结合活性将是 检查过了。结合的或非结合的糖蛋白将用于 金黄地鼠皮下和口服免疫及其免疫应答 将会被评估。使用基因融合的CTB和抗原的可能性 作为疫苗的分子也将在第二阶段进行检查。最后， 方法将扩展到其他副粘病毒，类型1和类型2，以 开发三价亚单位疫苗。

The long term objective of this proposal is to develop a commercial trivalent glycoprotein subunit vaccine for parainfluenza viruses. Because of the high cost of preparing these glycoproteins, a means for achieving an enhanced immune response is an important objective, and is the immediate focus of this proposal. It has been shown that cholera toxin subunit B (CTB) is not only a potent mucosal immunogen but also effective adjuvant for antibody responses to unrelated antigens coadministered. The adjuvanticity of CTB to the PI3 glycoprotein subunit vaccine will be evaluated. Two glycoproteins, fusion (F) and hemagglutin-neuraminidase (HN) proteins, of PI3 virus will be purified from virions or virus infected cells by affinity columns coupled with specific monoclonal antibodies against F or HN. Glycoproteins will be conjugated with CTB by various methods and antigenicity of each component of conjugates as well as the binding activity of conjugated CTB to GM1 ganglioside will be examined. Conjugated or unconjugated glycoproteins will be used for subcutaneous and oral immunization of hamsters and their immune responses will be evaluated. Possibility to use genetically fused CTB and antigen molecules as vaccines will be also examined in Phase II. Finally, the approach will be extended to other paramyxovimses, type 1 and 2, to develop trivalent subunit vaccine.