两个NIH小鼠种群对不同种类的乙肝疫苗免疫应答效应的比较

Comparison of the Potency Testing by Immune Responses to Different Kinds of HB Vaccines in Two Close-Colonies of NIH Mice

目的比较两种不同的NIH小鼠种群基因组DNA中H-2q单倍型的百分比,对重组基因乙型肝炎疫苗(酿酒酵母)、重组基因乙型肝炎疫苗(CHO)、重组基因乙型肝炎疫苗(汉逊酵母)和治疗性乙型肝炎疫苗(乙克)免疫应答的影响。方法检测乙肝疫苗免疫效力用酶标法,测定乙肝疫苗的ED50值,再通过微量细胞毒法和PCR方法检测小鼠的H-2单倍型,计算不同品系小鼠H-2q的百分数,得出乙肝疫苗与不同动物种群的相关性。结果经验证,经1号NIH小鼠检测结果证实,重组基因乙型肝炎免疫效力依次为CHO>汉逊>酿酒,疗性乙型肝炎疫苗(乙克)效力检测合格,但经2号NIH小鼠检测结果显示除CHO疫苗合格外,其他三种疫苗检测结果均为不合格。基因检测结果为,1号种群NIH鼠含有H-2q型基因的百分比是96%,而2号种群NIH小鼠含有H-2q型基因基因百分比为30%,因此不同的q型基因百分比导致了不同的检测结果。结论根据本研究结果表明虽然NIH鼠的H-2单倍型中有q型基因,但不同来源的小鼠种群之间会存在差异,经不同的环境饲养后,小鼠的基因会发生不同的变化,因此在进行生物制品检定时,要注意不同品系小鼠及同一品系小鼠不同种群之间的差异对试验本身的影响。建立一种"动物免疫"的概念。

Objective To evaluate the efficacy of immune responses to different kinds of recombinant HB （rHB） vaccines in different strains of mice. Metheods All the mice were injected HB vaccines in a serial dilution. Two NIH close-colonies of mice were typed in the H-2 region by using microcytotoxicity method and PCR. The titre of anti-HBsAg antibody was analyzed by microplate reading. The ED50 was then estimated by Karder method for each strain. Results Significant differences were found between potency estimates derived from assays using the two close-colonies of NIH mice. Conclusion It is likely that the variation of immune responses to HB vaccines in mice is correlated with the H-2 haplotype. In some particular cases, the heterezygosity in H-2 region found in NIH stock may influence the accuracy in such testing even a reference preparation of HB vaccine was used. Based on our findings, attention should be paid to select an appropriate NIH stock with the H-2q haplotype for potency testing of HB vaccine in China.