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目的 利用DHBV感染鸭模型 ,研究鸭IFN γ真核表达质粒作为免疫佐剂对DNA疫苗诱导免疫应答的影响和预防DHBV感染的作用。方法 用DuIFN γ真核表达质粒与DHBpreS SDNA疫苗共免疫 ,或用DHBpreS SDNA疫苗单独免疫正常幼鸭 ,检测经免疫前后鸭PBMC表达DuIFN γ的mRNA水平 (半定量竞争性RT PCR)、诱生的抗体水平 (ELISA)、病毒攻击免疫鸭后血清和肝脏中的病毒DNA变化 (斑点和Southern印迹核酸杂交 )。结果 IFN γ表达质粒作为免疫佐剂 ,可以增强DNA疫苗诱导的鸭PBMCIFN γ的mRNA表达水平。用大剂量病毒攻击免疫鸭的结果显示 ,用DuIFN γ表达质粒和DHBpreS SDNA疫苗共免疫鸭清除DHBV的速度 ,明显比仅用DHBpreS SDNA疫苗免疫鸭清除病毒的速度快 ,肝脏中DHBV总DNA和共价闭环DNA(cccDNA)的量也低于DHBpreS SDNA单独免疫组。结论 IFN γ真核表达质粒作为免疫调节佐剂在DNA免疫中具有潜在的应用价值。
Objective To study the effect of duck IFNγ eukaryotic expression plasmid as immune adjuvant on the immune response induced by DNA vaccine and the prevention of DHBV infection in ducks infected with DHBV. Methods DuIFN γ eukaryotic expression plasmids were co-immunized with DHBpreS SDNA vaccine or normal ducklings were immunized with DHBpreS SDNA vaccine alone. The expression of DuIFN γ mRNA in duck PBMCs was measured before and after immunization (semiquantitative competitive RT PCR) Antibody levels (ELISA), changes in viral DNA in sera and liver following challenge with virus challenge ducks (blot and Southern blot nucleic acid hybridization). Results IFNγ expression plasmid as an adjuvant for immunization could enhance DNA vaccine-induced duck PBMC IFNγ mRNA expression. Immunization of ducks with large doses of virus challenged the results of co-immunization of ducks with DuIFN [gamma] expression plasmid and DHBpreS SDNA vaccine to clear the rate of DHBV significantly faster than the DHPV vaccine alone DHV vaccination to clear the virus, DHBV total DNA in the liver and total The amount of closed loop DNA (cccDNA) was also lower than that of DHBpreS SDNA alone immunization group. Conclusion The IFNγ eukaryotic expression plasmid has potential value as an immunomodulatory adjuvant in DNA immunization.