栽培一枝蒿粗多糖混合口蹄疫疫苗乳化方法及稳定性分析

Analysis of Emulsifying Method and Stability of Foot-and-mouth Disease Vaccine Combined with Crude Polysaccharides from Cultivated Artemisia rupestris L.

旨在探究栽培一枝蒿粗多糖(cultivated Artemisia rupestris L.crude polysaccharides,CARCP)协同ISA-206油乳剂通过比较不同乳化方法与口蹄疫灭活抗原(inactivated foot-and-mouth disease viral antigen,FMD-Ag)制备FMD疫苗的效果异同,并观察不同储存条件对FMD疫苗稳定性的影响。通过搅拌和超声乳化分别制备不同疫苗组合,皮下免疫ICR小鼠后检测FMD特异性抗体及T细胞亚群比例。将搅拌乳化的不同疫苗组合在4、25℃条件下放置30、180 d后免疫ICR小鼠,检测体液和细胞免疫水平分析其稳定性。结果显示:与商品化FMD疫苗相比,搅拌和超声乳化制备的CARCP单独或协同ISA-206油乳剂的FMD疫苗组的特异性IgG水平和CD3^(+)CD4^(+)、CD3^(+)CD8^(+)T细胞比例无显著性影响(P>0.05)。4℃放置30、180 d后,与商品化FMD疫苗相比,CARCP单独或协同ISA-206油乳剂的FMD疫苗组的IgG水平和CD4^(+)、CD8^(+)、CD4^(+)CD44^(+)、CD8^(+)CD44^(+)T细胞比例无显著性差异(P>0.05)。25℃放置30、180 d后,与商品化FMD疫苗相比,免疫动物后FMD-Ag+ISA-206抗体水平及T细胞亚群比例显著降低(P<0.05),而FMD-Ag+ISA-206+CARCP抗体水平及T细胞亚群比例无显著性差异(P>0.05)。搅拌和超声乳化制备的不同FMD疫苗组合的抗体水平和T细胞水平的免疫效果没有差异;不同储存条件下,CARCP能够增强FMD疫苗的稳定性,这些研究结果为CARCP多糖佐剂的研究提供试验依据。

The study aimed to explore the efficacy of the different emulsification methods and stability of foot-and-mouth disease(FMD)vaccine adjuvanted with cultivated Artemisia rupestris L.crude polysaccharides(CARCP).ICR mice were subcutaneously immunized with FMD-Ag(inactivated foot-and-mouth disease viral antigen)adjuvanted with ISA-206 or ISA-206^(+)CARCP prepared by stirred emulsification and ultrasonic emulsification.FMD-specific antibodies and T cell subsets from spleen were detected.After FMD-Ag+ISA-206 and FMD-Ag+ISA-206^(+)CARCP by stirred emulsification were placed at 4℃and 25℃for 30 and 180 d,respectively,ICR mice were immunized with these vaccine formulations and tested humoral and cellular immunity for the analysis of stability.Results were as follows:Compared with commercial FMD vaccine,the effects of different FMD vaccine formulations prepared by stirred emulsification and ultrasonic emulsification had no significant difference on FMD-specific IgG and CD3^(+)CD4^(+)and CD3^(+)CD8^(+)T cells(P>0.05).After placing at 4℃for 30 and 180 d,compared with commercial FMD vaccine,there was no significant difference in IgG levels and CD4^(+),CD8^(+),CD4^(+)CD44^(+),CD8^(+)CD44^(+)T cells in FMD-Ag+ISA-206 and FMD-Ag+ISA-206^(+)CARCP group(P>0.05).After placing at 25℃for 30 and 180 d,compared with commercial FMD vaccine,the levels of antibodies and T-cell subsets in FMD-Ag+ISA-206 group were significantly reduced(P<0.05),while there was no significant difference between antibody levels and T-cell subsets in FMD-Ag+ISA-206^(+)CARCP group(P>0.05).Stirred and ultrasonic emulsification have similar immune effects on FMD vaccine.Under different storage conditions,CARCP enhanced the stability of FMD vaccine.These data provided some experimental thoughts for CARCP polysaccharide adjuvant in the development of FMD vaccine adjuvant.