狂犬病疫苗脂质体冻干粉的免疫原性评价

目的评价狂犬病疫苗脂质体冻干粉的免疫原性。方法将狂犬病疫苗原液稀释至15 IU/mL,制备为稀释液冻干粉(Y1组);将该稀释液冻干粉与脂质体冻干粉按5∶3的体积比混合,制成物理混合物(Y2组);同时采用冻融-冻干法将狂犬病疫苗稀释液和脂质体配制成狂犬病疫苗脂质体冻干粉(Y3组)。将各组疫苗复溶后,分别于0、3、7、14、21 d经小鼠腹腔给药,0. 5 mL/只。MTT法检测小鼠脾淋巴细胞增殖能力,流式细胞术检测T淋巴细胞表面标记,ELISA法测定小鼠体液中狂犬病病毒抗体IgG(RV-IgG)浓度。结果与Y1组及Y2组比较,Y3组初次免疫后3 d小鼠脾细胞刺激指数(stimulatingindex,SI)值明显升高(P <0. 01);初次免疫后7、14、21、28 d,CD4^+/CD8^+值明显升高(P <0. 05),初次免疫后7、14、21 d,RV-IgG浓度明显升高(P <0. 05)。结论狂犬病疫苗脂质体冻干粉在小鼠体内具有良好的免疫原性,脂质体能提高疫苗免疫原性,延长疫苗的免疫时间,有望开发成为新一代广泛应用的疫苗佐剂。

Immunogenicity of liposome lyophilized powder of rabies vaccine

Objective To evaluate the immunogenicity of liposome lyophilized powder of rabies vaccine. Methods The bulk of rabies vaccine was diluted to a concentration of 15 IU/mL and prepared into diluent lyophilized powder(Y1).The diluent lyophilized powder was mixed with liposome lyophilized powder at a volume ratio of 5 ∶ 3 to prepare a physical mixture(Y2). Meanwhile,rabies vaccine liposome lyophilized powder(Y3) was prepared with the diluent and liposome by freezing and thawing-lyophilization. The vaccines in various groups were reconstituted and injected i.p. into mice on days 0,3,7,14 and 21 separately,0. 5 mL for each. The proliferation abilities of splenic lymphocytes of the mice in various groups were determined by MTT assay,while the surface markers of T lymphocytes by flow cytometry,and humoral anti-rabies virus IgG(RV-IgG)levels by ELISA. Results Compared with those in Y1 and Y2 groups,the stimulating index(SI)of splenocytes of mice in Y3 group 3 d after the first immunization increased significantly(P < 0. 01),while the CD4^+/CD8^+ ratio 7,14,21 and 28 d as well as the RV-IgG level 7,14 and 21 d after the first immunization increased significantly(P < 0. 05). Conclusion The rabies vaccine liposome lyophilized powder showed good immunogenicity in mice. The liposome enhanced the immunogenicity and prolonged the time duration of immunity of the vaccine,which was expected to be a widely used novel adjuvant.