弗氏佐剂乳化抗原不同注射体积与小鼠鼓包结痂形成及免疫原性的关系

Relationship Between Different Injection Volumes of Freund's Adjuvant Emulsified Antigen and Tympanoplasty and Immunogenicity in Mice

为比较小鼠背部弗氏佐剂乳化抗原不同注射体积与结痂形成情况及免疫原性产生的关系。按照确定的免疫程序,以公司自制2019nCOV-SRBD蛋白与弗氏佐剂乳化,每组分别于小鼠背部各点注射不同体积,定期观察小鼠背部鼓包结痂形成情况,用间接ELISA法检测抗2019nCOV-SRBD特异性抗体效价,t检验统计分析数据。结果:在1点注射组、2点注射组、5点注射组每只小鼠注射部位均有不同程度的鼓包。1点注射组鼓包最明显;2点注射组一侧消失,一侧略有鼓包;5点注射组鼓包不明显。3组ELISA效价GMT值依次为32254、16127、51200,5点注射组GMT值最高,1点注射组GMT值最低,但三者之间进行T检验,均无显著性差异(P<0.05)。结论:弗氏佐剂作为免疫增强剂需多点注射,每点注射体积为0.1~0.2 mL,既能减少动物机体不适,保证动物伦理福利,又能达到试验预期。

The purpose of this study was to compare the effects of different injection volumes of freund's adjuvant on the formation of scab and immunogenicity in mice.The 2019nCOV-SRBD protein was emulsified with freund's adjuvant in accordance with the established immune procedure.Different volumes were injected in each group at different points on the back of mice,and the swelling and scab formation on the back of mice were observed regularly,the titer of specific antibody against 2019nCOVSRBD was detected by indirect Elisa,and the data were analyzed by t test.Results:In 1 point injection group,2 point injection group,5 point injection group,each mouse injection site has different degrees of swelling.The bulge was most obvious in the 1 point injection group,disappeared in the 2 point injection group and had a slight bulge on one side,and was not obvious in the 5 point injection group.The GMT values of the three groups were 32254,16127 and 51200,respectively.The GMT values of the five-point injection group were the highest and the GMT values of the one-point injection group were the lowest,but there was no significant difference among the three groups by T test(P<0.05).Conclusion:Freund's adjuvant,as an immunopotentiator,should be injected in multiple points with the volume of 0.1~0.2 mL per point,which can not only reduce the discomfort of animals,but also ensure the ethical welfare of animals.