口蹄疫疫苗非抗原蛋白对146S抗原免疫效果的影响

为探究非抗原蛋白对口蹄疫病毒A型(FMDV-A) 146S抗原免疫效果的影响,本研究以小鼠和猪为试验动物,分别制备5组小鼠注射用样品:A组(3μg 146S抗原)、B组(3μg 146S抗原+25μg非抗原蛋白)、C组(3μg 146S抗原+50μg非抗原蛋白)、D组(3μg 146S抗原+100μg非抗原蛋白)及E组(空白对照,PBS);同时制备4组猪注射用样品:A组(18μg 146S抗原)、B组(18μg 146S抗原+400μg非抗原蛋白)、C(18μg 146S抗原+4 000μg非抗原蛋白)及D组(空白对照,PBS)。免疫接种试验动物后,通过淋巴细胞增殖试验及实时荧光定量PCR评价比较不同疫苗样品组小鼠产生的细胞免疫应答水平,同时通过液相阻断ELISA方法检测、评价各试验组动物(小鼠和猪)产生的体液免疫应答水平。小鼠试验结果显示,在免疫后3个检测时间点内,4个试验组中C组的平均抗体水平最高,平均抗体效价分别为4.13、5.83和5.50,而D组的抗体水平最低,平均抗体效价分别为3.46、5.16和4.46;C组细胞增殖能力及Th-1型细胞因子IL-6、IFN-β和TNF-αmRNA表达量均高于其他组。猪试验结果显示,A组和B组间平均抗体水平差异不显著(P>0.05),但A组和B组的平均抗体效价均极显著高于C组(P<0.01)。综合上述结果表明,一定范围内少量非抗原蛋白对FMDV-A 146S抗原的免疫效果没有影响,而高浓度的非抗原蛋白则抑制FMDV-A 146S抗原的免疫效果。

Effect of Non-antigen Protein of Foot-and-Mouth Disease Vaccine on Immune Effect of 146S Antigen

In order to explore the effect of non-antigen protein on the immune effect of foot-and-mouth disease virus type A (FMDV-A) 146S antigen,mice and pigs were used as experimental animals.5 groups of mice for injection were prepared:Group A (3 μg 146S antigen),group B (3 μg 146S antigen＋25 μg non-antigenic protein),group C (3 μg 146S antigen＋50 μg non-antigenic protein),group D (3 μg 146S antigen＋100 μg non-antigenic protein) and group E (blank control,PBS).At the same time,four groups of pig injection samples were prepared:Group A (18 μg 146S antigen),group B (18 μg 146S antigen＋400 μg non-antigenic protein),group C (18 μg 146S antigen＋4 000 μg non-antigenic protein) and group D (blank control,PBS).The cell immune response levels of mice in different vaccine sample groups were compared by lymphocyte proliferation assay and Real-time PCR.The level of humoral immune response produced by the animals (mice and pigs) of each test group was detected and evaluated by a liquid phase blocking ELISA method.The results of mice test showed that the average antibody level of group C was the highest in four test groups at the three test time points after immunization,and the average antibody titers were 4.13,5.83 and 5.50,respectively,while the antibody of group D was the lowest,the average antibody titer average antibody titers were 3.46,5.16 and 4.46 in the antibody detection,respectively;The cell proliferation ability and Th-1 type cytokine IL-6,IFN-β and TNF-α mRNA expression in group C were higher than other groups.The results of pigs test showed that the average antibody level between groups A and B was not significantly different (P>0.05),but the average antibody titers of groups A and B were extremely significantly higher than that of group C (P<0.01).The above results indicated that a small amount of non-antigenic protein had no effect on the immune effect of FMDV-A 146S antigen within a certain range,while a high concentration of non-antigenic protein inhibited the immune effect of FMDV-A 146S antigen.