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病毒性载体疫苗是一种有前途的艾滋病疫苗。为了构建以重组腺相关病毒8型(recombinant adeno associated virus type 8,rAAV8)为载体,表达HIV或SIV包膜蛋白的艾滋病疫苗。同时在小鼠体内对其免疫原性进行评价,为下一步研究奠定基础。本研究分别构建了表达HIV AE亚型和SIV mac239株包膜蛋白(不含胞内区)的rAAV8-AEgp145和rAAV8-SIVenvT两种重组病毒,并通过PCR和Western Blot方法对重组病毒进行了体外鉴定。将两种重组病毒分别接种BALB/c小鼠,应用ELISA和ELISPOT方法检测小鼠的HIV/SIV特异性抗体滴度和细胞免疫应答强度。结果显示,rAAV8能够在293T细胞中高效表达HIV AEgp145和SIV envT基因。小鼠接种两种重组病毒3-5W后,均能检测到gp120特异性抗体和env特异性细胞免疫应答,并且在16-20W后反应强度仍显著高于对照组。以上结果提示,携带HIV AEgp145和SIV envT基因的rAAV8载体能够在小鼠体内诱导中等强度并且持续时间较长的特异性体液和细胞免疫应答。
Abstract：Viral vector-based vaccines are considered to be promising for acquired immunodeficiency syndrome(AIDS) patients. We wished to construct recombinant adeno associated virus type 8(rAAV8) expressing human immunodeficiency virus(HIV)-AEgp145 or simian immunodeficiency virus(SIV)-envT gene,to evaluate immunogenicity of these two vaccines in a mouse model and lay the foundation for further research.These two genes were constructed into the rAAV8 vector respectively. rAAV8-AEgp145 and rAAV8-SIVenvT were identified in vitro via PCR and Western Blotting. BALB/c mice were immunized with two recombinant viruses. The HIV/SIV gp120 specific antibody titer and cellular immune response level were measured by ELISA and ELISpot,respectively. We discovered that rAAV8-AEgp145 and rAAV8-SIVenvT were expressed effectively in 293 T cells. HIV/SIV gp120-specific antibody and Env-specific cellular immunology could be detected 3-5 weeks post immunization. The immune response in the immunization group lasted for 16-20 weeks. In conclusion,rAAV8-AEgp145 and rAAV8-SIVenvt induced expression of a high level of HIV/SIV-specific antibody and a moderate cellular immune response in mice.
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基本信息:
DOI：10.13242/j.cnki.bingduxuebao.003971
中图分类号:R392
引用信息:
[1]何小周,杨靖,李红霞,等.重组HIV-AEgp145和SIV-envT基因的rAAV8在小鼠体内的免疫原性研究[J].病毒学报,2021,37(03):613-620.DOI:10.13242/j.cnki.bingduxuebao.003971.
基金信息:
艾滋病和病毒性肝炎等重大传染病防治科技重大专项(项目号：2018ZX10731-101-002-010);题目：MVA及Ad5载体疫苗联合应用的临床前研究~~
2020-12-14
2020
2021-02-19
2021-05-12
2021
1
2021-05-14
2021-05-14
2021-05-14