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目的比较包裹乙型肝炎(简称乙肝)的壳聚糖纳米粒与常规铝佐剂乙肝疫苗的免疫效果差异,并对壳聚糖佐剂的应用效果进行评价。方法采用离子交联法制备20、30及40μg/ml的包裹乙肝疫苗的壳聚糖纳米粒溶液,激光粒度分析仪测定纳米粒粒径和多分散系数,BCA法测定包封率;将BALB/c小鼠随机分为4组:铝佐剂对照组、空白纳米粒对照组、常规疫苗组和壳聚糖疫苗组,肌肉注射免疫小鼠,2μg/只,隔周免疫1次,共2次。采用全自动免疫发光分析仪对小鼠血清中乙肝表面抗体进行定量分析。结果制备的载乙肝疫苗壳聚糖纳米颗粒粒径为(262.7±10.8)nm,与空白壳聚糖纳米粒粒径相比,略有减小,多分散系数为(0.228±0.016);随着抗原浓度升高、4℃放置3个月及反复冻融,纳米粒的包封率均未发生显著变化,可达80%左右;壳聚糖疫苗组在小鼠体内产生的表面抗体滴度最高可达1∶80 000以上,并从第8周起,与常规疫苗组间差异有统计学意义(P<0.01)。结论壳聚糖纳米粒作为新型疫苗佐剂,具有一定的应用价值和进一步研究的意义。
OBJECTIVE: To compare the immune effects of chitosan nanoparticles encapsulated with hepatitis B (hepatitis B) and conventional aluminum adjuvant hepatitis B vaccine and to evaluate the application effect of chitosan adjuvants. Methods 20, 30 and 40 μg / ml chitosan nanoparticles solution were prepared by ion-crosslinking method. The particle size and polydispersity of nanoparticles were measured by laser particle size analyzer. The encapsulation efficiency was determined by BCA method. The BALB / c mice were randomly divided into 4 groups: aluminum adjuvant control group, control group blank nanoparticles, conventional vaccine group and chitosan vaccine group, intramuscular immunized mice, 2μg / only, immunized every other week for 2 times . Quantitative analysis of hepatitis B surface antibody in mouse serum was performed by using automatic immune luminescence analyzer. Results The diameter of chitosan nanoparticles loaded with hepatitis B vaccine was (262.7 ± 10.8) nm, which was slightly lower than that of chitosan nanoparticle. The polydispersity index was (0.228 ± 0.016) The antigen concentration increased, placed at 4 ℃ for 3 months and repeated freezing and thawing, the encapsulation efficiency of nanoparticles did not change significantly, up to about 80%; chitosan vaccine group produced the highest surface antibody titers in mice Up to more than 1: 80000, and from the 8th week, the difference was statistically significant (P <0.01) with the conventional vaccine group. Conclusion Chitosan nanoparticles as a new vaccine adjuvant, has a certain value and further research significance.