利用中国流行株HIV-1 Gag重组痘苗病毒与核酸疫苗免疫的新程序

目的 将中国流行株ＨＩＶ－１　Ｇａｇ基因在痘苗病毒中表达，以重组痘苗病毒与核酸疫苗混合免疫，进 而研制艾滋病疫苗。方法 将中国流行株ＨＩＶ－１ Ｇａｇ基因片段插入到ｐＪ３８载体启动子下游，经同源重组和血凝素 阴性空斑筛选重组痘苗病毒，经 ＳＤＳ－ＰＡＧＥ和 Ｗｅｓｔｅｒｎ检测目的蛋白。以重组痘苗病毒与核酸疫苗进行小鼠免 疫试验，淋巴细胞转化实验，ＣＩＬ，ＣＤ＋４；ＣＤ＋８细胞计数及细胞免疫和体液免疫水平检测。结果 重组痘苗病毒ｖＪ３８ Ｇａｇ具有良好的免疫原性，与２ｒＶＶ－ＤＮＡ混合免疫效果更好。结论 重组痘苗病毒 ｖＪ３８ Ｇａｇ能够表达Ｇａｇ蛋白并诱 导机体产生细胞免疫和体液免疫。

A Novel Combined Immunization Schedule of Recombinant Vaccinia Virus Containing Gag Gene of HIV-1 Subtype B and DNA Vaccine

Objective To express the Gag gene of HIV-1 subtype B in vaccinia virus and evaluate the effect of combined immunization with the prepared recombinant vaccinia virus and DNA vaccine. Methods The Gag gene of HIV-1 subtype B was inserted to the downstream of promoter of pJ38 vector, then recombinant vaccinia virus was screened out by homologous recombination and plaque assay. The expressed plaque was examined by SDS-PAGE and Western blot. The recombinant vaccinia virus was used for immunizing mice combined with DNA vaccine,and the effect was evaluated by lymphocyte transformation test, CTL, CD4 and CD8 T cell counting as well as cellular and humoral response levels. Results The recombinant vaccinia virus vJ38 Gag showed good immunogenicity. The optimal schedule was two doses of recombinant virus vJ38 Gag( primary immunization) plus one dose of DNA vaccine( booster) . Conclusion Gag protein is expressed in recombinant virus vJ38Gag, and both cellular and humoral immune responses were induced.