论文部分内容阅读
目的评估沙门氏菌鞭毛蛋白作为添加剂在氢氧化铝疫苗制剂中的作用。方法以卵清蛋白(OVA)为抗原,氢氧化铝(Alhydrogel)为佐剂,鼠伤寒沙门氏菌鞭毛蛋白(FliC)或寡聚脱氧核糖核苷酸CpG-ODN 1826为添加剂,制备OVA/Alhydrogel、OVA/Alhydrogel+FliC和OVA/Alhydrogel+CpG-ODN 1826疫苗制剂。BALB/c小鼠随机分成3组,每组6鼠,分别用以上疫苗制剂以肌肉注射的方式免疫2次,间隔28d。在初次免疫后第28d和42d采小鼠血,分离血清,用ELISA法测定血清中抗OVA IgG及亚型。在初次免疫后的第42d处死小鼠,取脾,制备脾细胞,给予抗原刺激后检测培养上清中γ干扰素(IFN-γ)和白介素4(IL-4)水平。结果初次免疫后第42d,OVA/Alhydrogel+FliC制剂组OVA/Alhydrogel+CpG-ODN 1826制剂组和OVA/Alhydrogel制剂组特异IgG滴度分别为(714.75±213.32)U-nit、(863.78±355.05)Unit和(267.45±8.49)Unit,差异有统计学意义(P<0.05);前两制剂组比较差异无统计学意义(P>0.05)。IgG亚型分析和脾细胞培养上清IFN-γ和IL-4水平测定表明,OVA/Alhydrogel制剂诱导Th2偏向的免疫应答,而在加入沙门氏菌鞭毛蛋白后免疫应答有向Th1/Th2混合型转变的趋势。结论沙门氏菌鞭毛蛋白能增强特异性免疫应答反应,有望成为氢氧化铝疫苗制剂的添加剂。
Objective To evaluate the role of Salmonella flagellin as an additive in aluminum hydroxide vaccine formulations. Methods OVA / Alhydrogel and OVA were prepared using ovalbumin (OVA) as antigen, Alhydrogel as adjuvant, Salmonella typhimurium flagellin (FliC) or oligodeoxyribonucleotide CpG-ODN 1826 as additives / Alhydrogel + FliC and OVA / Alhydrogel + CpG-ODN 1826 vaccine formulations. BALB / c mice were randomly divided into 3 groups, 6 mice in each group. The mice were immunized twice intramuscularly with the above vaccine preparations for 28 days. At the 28th day and the 42th day after the first immunization, the blood of mice was taken and the serum was separated. The serum anti-OVA IgG and its subtype were measured by ELISA. The mice were sacrificed on the 42th day after the primary immunization, spleen cells were taken, spleen cells were prepared, and the levels of IFN-γ and IL-4 in the culture supernatants were detected after antigen stimulation. Results The specific IgG titers of OVA / Alhydrogel + CpG-ODN 1826 group and OVA / Alhydrogel group were (714.75 ± 213.32) U-nit and (863.78 ± 355.05) Unit and (267.45 ± 8.49) Unit, the difference was statistically significant (P <0.05); there was no significant difference between the two preparations (P> 0.05). Analysis of IgG subtypes and determination of IFN-γ and IL-4 levels in splenocyte culture supernatants showed that the OVA / Alhydrogel preparations induced a Th2-biased immune response, whereas the immune response to the Th1 / Th2 mixed-type transition after addition of Salmonella flagellin trend. Conclusion Salmonella flagellin can enhance the specific immune response and is expected to become an additive of aluminum hydroxide vaccine.