摘要
目的构建表达Ⅰ型艾滋病病毒(human immunodeficiency virus-1type,HIV-1)CN54膜蛋白Gp145去糖基化突变体的DNA疫苗和重组痘病毒疫苗,并对其进行抗原改造和免疫原性测试。方法采用分子生物学方法构建了表达HIV-1CN54膜蛋白Gp145或其突变体(Gp145dG,去除了V1/V2区的6个糖基化位点)的DNA疫苗(SV145、SV145dG)和重组痘病毒疫苗(rTTV145、rTTV145dG)。运用DNA初免-重组痘病毒疫苗加强的方式接种雌性BALB/c小鼠,免疫结束后,分别采用IFN-γ酶联免疫斑点技术(enzyme-linked immunospot assay,ELISPOT)和酶联免疫吸附测定技术(enzyme-linked immunosorbent assay,ELISA)对Gp145特异性细胞免疫反应和抗体反应进行检测。结果 SV145初免-rTTV145加强所活化的特异性T细胞反应强度为(1 949±1 307)斑点形成细胞数(spotforming cells,SFCs)/106个脾淋巴细胞,对数抗体滴度为(4.020±0.346);SV145dG初免-rTTV145dG加强所活化的特异性T细胞反应强度为(1 192±540)SFCs/106个脾淋巴细胞,对数抗体滴度为(3.300±0.298)。结论表达HIV-1CN54膜蛋白Gp145的DNA疫苗和重组痘病毒疫苗具有良好的免疫原性,去除V1/V2区的糖基化位点无助于提高其免疫原性。
Objective To construct DNA and recombinant Tiantan vaccinia vaccines encoding the deglycoslated mutant of HIV-1CN54 Gp145 and to investigate the immunogenicity.Methods Molecular biology techniques were used.DNA（SV145、SV145dG） and recombinant Tiantan vaccinia（rTTV145、rTTV145dG） vaccines encoding wild type or V1 / V2 deglycosylated Gp145 were constructed.Female BALB / c mice were then immunized,the T cell and antibody responses were detected by using IFN-γ ELISPOT and ELISA,respectively.Results Specific T cell responses elicited by SV145 prime-rTTV145 was（1 949 ± 1 307） SFCs /106 splenocytes,the log titer of specific binding antibody was（4.020 ± 0.346）;specific T cell responses elicited by SV145dG prime-rTTV145dG was（1 192 ± 540） SFCs /106 splenocytes and the log titer of specific binding antibody was（3.300 ± 0.298）.Conclusions Both the DNA and recombinant Tiantan vaccinia vaccines encoding HIV-1CN54 Gp145 are immunogenic.However,V1 / V2 deglycosylation didn’t help to further improve its immunogenicity.
出处
《中华疾病控制杂志》
CAS
北大核心
2013年第6期461-464,共4页
Chinese Journal of Disease Control & Prevention
基金
"十二五"国家重大科技专项(2012ZX10001008
2012ZX10004501-001-006)
国家自然科学基金(81020108030
31100126)
传染病预防控制国家重点实验室课题(2012SKLID103
2011SKLID207)
上海市自然科学基金(11ZR1430600)
