W/O/W佐剂添加两种免疫增强剂对口蹄疫灭活疫苗的影响OA北大核心CSTPCD
Effect of W/O/W adjuvant added with two immunopotentiator on inactivated foot-and-mouth disease vaccine
为进一步提高口蹄疫灭活疫苗的免疫效果,本研究在水包油包水(W/O/W)佐剂配方上添加免疫增强剂,一种是天然化合物维生素E(VE),另一种是人工合成的咪喹莫特衍生物(IMQ-Chol).每种免疫增强剂设低中高三种不同剂量,加入到基础配方中制备成不同的佐剂,然后与口蹄疫灭活病毒进行配伍.观察各组疫苗的外观并检测其稳定性、黏度、粒径和多分散系数(PDI)以确定其理化性质;称量小鼠体重、检测血常规、血液生化和组织学变化来评估免疫增强剂的安全性;检测小鼠血清中的特异性抗体、抗体亚型、细胞因子水平以及小鼠脾T淋巴细胞的分化情况,评价免疫增强剂对疫苗免疫效果的影响.结果显示,配制的疫苗均为乳白色W/O/W乳液,37 ℃放置9周疫苗未出现分层现象,黏度均在16 cp以下,粒径在270 nm～370 nm之间,PDI在0.34～0.42之间;不同剂量的免疫增强剂对小鼠体重没有影响(P＞0.05),血常规、血液生化及组织学切片结果表明两种免疫增强剂的生物安全性良好;添加VE或IMQ-Chol的免疫组与基础配方免疫组相比,特异性抗体水平均显著提高(P＜0.01);其中,添加高剂量VE的免疫组和添加低剂量IMQ-Chol的免疫组不仅特异性抗体水平显著提高(P＜0.001),Th1型相关分子(IgG2a、IFN-γ、IL-12)和Th2型相关分子(IgG1、IL-4、IL-6)的分泌水平也有提升(P＜0.05);这两组的CD3+CD4+T细胞的比例分别为75.2％和77.3％,高于基础配方组的69.6％,且CD4+/CD8+比值也高于基础配方组(P＜0.05).以上结果表明,VE和IMQ-Chol这两种免疫增强剂在不影响疫苗的理化特性的同时都具有良好的安全性,并且添加高剂量的VE或低剂量的IMQ-Chol更能有效地提高口蹄疫灭活疫苗的免疫效果.
To enhance the immune efficacy of the inactivated foot-and-mouth disease vaccine,this study incorporated immunopotentiators into the water-in-oil-in-water(W/O/W)adjuvant formulation.Two types of immunopotentiators,natural compound vitamin E(VE)and synthetic derivative imiquimod(IMQ-Chol)were used.The basic formulation was mixed with the inactivated foot-and-mouth disease virus,along with three different doses(low,medium,high)of each immunopotentiator,to prepare various adjuvants.The appearance and physicochemical properties of each vaccine were determined,including stability,viscosity,particle size,and polydispersity index(PDI).The safety of the immunopotentiator was evaluated through mouse body weight,routine blood test,blood biochemistry,and histological changes.Furthermore,the impact of the immunopotentiator on vaccine immune efficacy was assessed by examining specific antibodies,antibody subtypes,cytokine levels in mouse sera,and differentiation of mouse splenic T lymphocytes.The results showed that the prepared vaccines were milky white W/O/W emul-sions with no phase separation after 9 weeks at 37 ℃.The viscosity remained below 16 cp,and the parti-cle size ranged from 270 nm to 370 nm,with the PDI between 0.34 and 0.42.None of the immunopotentiators affected mouse body weight(P＞0.05),and routine blood test,blood biochemistry and histological sections indicated that both immunopotentiators had good biological safety.Compared to the basic formula group,the groups immunized with VE or IMQ-Chol showed significantly higher level of specific antibody(P＜0.01).Notably,the group with high-dose VE and low-dose IMQ-Chol significantly increased the spe-cific antibody level(P＜0.001),as well as the secretion levels of Th1-related molecules(IgG2a,IFN-γ,IL-12)and Th2-related molecules(IgG1,IL-4,IL-6)(P＜0.05).The proportions of CD3+CD4+T cells in these two groups were 75.2％and 77.3％,which were higher than the 69.6％in the basic formulation group,and the CD4+/CD8+ratio was higher as well(P＜0.05).The above results indicated that both VE and IMQ-Chol did not impact the physicochemical properties of the vaccine,maintained good safety,and the addition of high-dose VE or low-dose IMQ-Chol could enhance the immune efficacy of the inactivated foot-and-mouth disease vaccine.
周宇杰;卢宇;许泽玉;唐健中;钱莺娟;邓碧华
南京农业大学动物医学院教育部动物健康与食品安全国际合作联合实验室江苏省外国专家工作室,江苏南京 210095||江苏省农业科学院动物免疫工程研究所,江苏南京 210014江苏省农业科学院动物免疫工程研究所,江苏南京 210014||兽用生物制品(泰州)国泰技术创新中心,江苏泰州 225300||江苏大学药学院,江苏镇江 212013南京农业大学动物医学院,江苏南京 210095南京农业大学动物医学院教育部动物健康与食品安全国际合作联合实验室江苏省外国专家工作室,江苏南京 210095江苏省农业科学院动物免疫工程研究所,江苏南京 210014||兽用生物制品(泰州)国泰技术创新中心,江苏泰州 225300
畜牧业
口蹄疫W/O/W佐剂免疫增强剂维生素E咪喹莫特
foot-and-mouth diseaseW/O/W adjuvantimmunopotentiatorvitamin Eimiquimod
《中国兽医科学》 2024 (003)
300-307 / 8
国家重点研发揭榜挂帅项目(2021YFD1801200);江苏省农业科技自主创新资金项目[CX(22)1011];江苏省重点研发计划项目(BE2021351)
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