蓝舌病毒1型灭活疫苗的制备与免疫效果

Preparation and immune effect of inactivated bluetongue virus type 1 vaccine

比较了二乙烯亚胺(BEI)、甲醛和β-丙内酯(BPL)3种灭活剂对蓝舌病毒1型(BTV-1)的灭活效果,利用免疫印迹法检测灭活病毒蛋白,选择BEI作为疫苗灭活剂制备BTV-1灭活疫苗并免疫绵羊,通过竞争ELISA法和噬斑减少中和试验(PRNT)测定抗体效价评价疫苗免疫效果。结果显示,BEI、BPL和甲醛均能在48 h内完全灭活BTV-1,且在BPL和BEI灭活处理的病毒样品中均可以检测到VP6、VP7、NS1和NS2四种病毒蛋白,而在甲醛处理的病毒样品中仅检测到了VP7蛋白,表明BEI和BPL在灭活病毒过程中对病毒蛋白抗原影响较小。血清抗体检测显示,与对照组相比,疫苗免疫组绵羊血清中抗BTV非中和抗体和中和抗体均显著升高。本研究制备的BTV-1灭活疫苗具有很好的免疫原性,这也为研制其他血清型蓝舌病疫苗奠定了基础。

We compared the inactivation effect on bluetongue virus type 1(BTV-1)by three inactivating agents including binary ethyleneimine(BEI),formaldehyde andβ-propiolactone(BPL).The inactivated viral proteins were confirmed by Western-blot,and BEI was selected as an inactivating agent to prepare inactivated BTV-1 vaccine and then immunize sheep.The antibody titers were tested by competitive ELISA and the plaque reduction neutralization test(PRNT)for evaluating the immune effect of the vaccine.The results showed that BEI,BPL and formaldehyde inactivated completely BTV-1 within 48 h,and BTV VP6,VP7,NS1 and NS2 proteins were detected in both BPL and BEI inactivated virus samples,but VP6,NS1 and NS2,except for VP7 protein,were not detected in the formaldehyde inactivated virus samples,which indicated that the BEI and BPL had little or no effect on the viral proteins during inactivation of the virus.Serum antibody tests showed that both non-neutralizing and neutralizing anti-BTV anti-bodies were significantly increased after immunization,compared with that of the control group.The BTV-1 inactivated vaccine prepared in this study has strong immunogenicity,which also lays a foundation for the development of other BTV serotype inactivated vaccines.