HIV-1中国流行株gag与hIL-2在重组痘苗病毒中的共表达及其与核酸疫苗的实验免疫研究

Coexpression of HIV-1 gag gene and hIL-2 gene in recombinant vaccinia viruses andits immunological study with HIV-1 DNA vaccine

目的：将HIV-1中国流行株B亚型gag基因、gag和hIL-2基因在天坛株痘苗病毒中进行共表达，以期获得重组痘苗病毒，观察细胞因子的佐剂作用，与核酸疫苗混合免疫，评价免疫效果，为新型艾滋病疫苗研制开发打下基础。方法：将HIV-1中国流行株 gag基因、gag和hIL-2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，SDS-PAGE、Western blot检测目的蛋白。以重组病毒和核酸疫苗免疫Balb/c小鼠，进行淋巴细胞转化实验、CTL、CD4+、CD8+ T细胞数目以及血清抗体的细胞免疫和体液免疫指标检测。结果：获得了重组痘苗病毒 vJ38gag和 vJ38gag-IL-2。与 vJ38-gag相比，vJ38gag-IL-2，具有更好的免疫原性，重组痘苗病毒免疫3次的效果好于重组病毒免疫2次，以2rVV-DNA混合免疫效果最好。结论：重组痘苗病毒vJ38gag和vJ38gag-IL-2能够表达外源蛋白并诱导机体产生细胞免疫和体液免疫。细胞因子IL-2发挥了免疫佐剂的作用。

Abstract Objective:To obtain recombinant vaccinia viruses expressed HIV-lcn subtype B gag and gag-hIL-2 chimera gene,evaluate theimmune effects in experimental mice by immunization with HIV-1 DNA vaccine,observe the immune adjuvant effect of IL-2.Metbods: HIV0-1gag and gag-hIL-2 chimera gene were inserted downstream of the combined promotor of pJ38 vector respectively and recombinant vaccinia virus-es were selected by plaque essay. Expression products were examined by SDS-PAGE and Western blot.Immune response of immunized Balb/cmice was evaluated by lymphocyte transformation test,CTL quantity change of T lymphocyte of CD4+、CD8+ and the level of serum antibody.Re-sults:Recombinant vaccinia viruses vJ38gag and vJ38gag-IL-2 were obtained and the expression were confirmed by SDS-PAGE and Western-blot. Recombinant vaccinia viruses show good immune effect when immuned with HIV-1 DNA vaccine.The best strategy for immunization is therVV-primed and HIV-1 DNA vaccine-boosted group and threer times of immunization were suggested to get effective results compared withvJ38gag an incressed immune response was induced by vJ38gag-IL-2. Conclusion: HIV-1 Gag protein and IL-2 protein can be expressed invJ38gag and vJ38gag-IL-2 respectively. IL-2 show good effect to ha sued as immune adjuvant.