重组幽门螺杆菌尿素酶B亚单位鼻腔免疫凝胶剂的研制

Preparation of Intranasal Immunization Gel Containing Recombinant Helicobacter Pylori Urease B

目的研制重组幽门螺杆菌尿素酶B亚单位疫苗凝胶剂,考察其特性及免疫效果。方法以卡泊波为基质,脱氧胆酸钠为黏膜吸收促进剂,制备凝胶剂,以落球法测定凝胶的黏度,SDS-PAGE电泳分析脱氧胆酸钠对蛋白纯度的影响,以免疫印记检测疫苗蛋白的完整性,以留样观察法及离心法考察凝胶的稳定性,以含抗原凝胶剂鼻腔免疫BALB/c小鼠,并与溶液剂相比较,考察凝胶剂的免疫效果。结果制备的凝胶剂稳定性好,凝胶的黏度为8050mPa.s,脱氧胆酸钠不影响蛋白的纯度,抗原蛋白的完整性良好。小鼠鼻腔免疫凝胶剂后产生了较强的特异性血清IgG及胃肠sIgA反应,优于液体免疫。结论重组幽门螺杆菌尿素酶B亚单位凝胶剂性质稳定,具有生物黏附性,可提高免疫效果,是较好的鼻腔免疫剂型。

OBJECTIVE To prepare Gel vaecine Containing recombine Helicobacter pylori UreB （rUreB）and study it＇s physical properties and immune efficiency by nasal route. METHODS The gel was prepared by carbopol. Sodium deoxycholate was applied to help the absorption of drug through mucosa. The Viscid coefficient and stability of gel were studied. The antigenic integrity was evaluated by western blotting. The immune efficiency was evaluated by intranasal immunization in BALB/c mice. RESULTS A stable gel was prepared with Viscid coefficient of $050 mPa · s. The results of western blotting showed that rUreB contained in gel was stable. The rUreB gel can induced a stronger antibody response （ IgA and IgG） than that of rUreB solution. CONCLUSION Stable rUreB gel is a good preparation to enhance immune efficiency by intranasal immunization.