The recombinant pseudorabies virus expressing African swine fever virus CD2v protein is safe and effective in mice.
The recombinant pseudorabies virus expressing African swine fever virus CD2v protein is safe and effective in mice.
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DOI：
10.1186/s12985-020-01450-7
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发表时间：
2020-11-16
期刊：
影响因子：
4.8
通讯作者：
Cai S
中科院分区：
文献类型：
--
作者：
Feng Z;Chen J;Liang W;Chen W;Li Z;Chen Q;Cai S
非洲猪瘟（African swine fever，ASF）是由非洲猪瘟病毒（African swine fever virus，ASFV）引起的一种传染病，可导致家猪和野猪的高死亡率。目前，没有商业疫苗可用于预防，疫情仍在蔓延。本研究构建了表达ASFV CD 2 v蛋白的重组伪狂犬病病毒（PRV-ΔgE/ΔgI/ΔTK-（CD 2 v）），并评价了其作为疫苗候选株的有效性和安全性。合成含有ASFV CD 2 v的同源重组片段并共转染HEK 293 T细胞，这是靶向PRV TK基因的敲除载体。用PRV-ΔgE/ΔgI感染转染细胞，在Vero细胞中进行噬斑纯化，获得重组株PRV-ΔgE/ΔgI/ΔTK-（CD 2 v）。重组病毒经测序、Western blotting和免疫荧光分析证实表达了ASFV CD 2 v，并在Vero细胞中传代20代以上检测了其遗传稳定性。在小鼠模型中进一步检测重组病毒的毒力、免疫原性和保护能力。PRV-ΔgE/ΔgI/ΔTK-（CD 2 v）重组株在Vero细胞中稳定，CD 2 v的加工不依赖于ASFV感染。PRV-ΔgE/ΔgI/ΔTK-（CD 2 v）疫苗接种既不引起瘙痒，也不引起全身性感染和炎症（具有白细胞介素-6（IL 6）的高表达）。此外，该病毒疫苗接种可产生抗CD 2 v特异性抗体并激活特异性细胞免疫应答，对强毒株（PRV-Fa）的攻击具有100%的保护作用。重组病毒接种后，解毒发生得更早，解毒量也低得多。PRV-ΔgE/ΔgI/ΔTK-（CD 2 v）重组株免疫原性强，安全有效，有望成为预防ASF和伪狂犬病的候选疫苗。
African swine fever (ASF) leads to high mortality in domestic pigs and wild boar and is caused by the African swine fever virus (ASFV). Currently, no vaccine is commercially available for prevention, and the epidemic is still spreading. Here, we constructed a recombinant pseudorabies virus (PRV) (PRV-ΔgE/ΔgI/ΔTK-(CD2v)) that expresses the CD2v protein of ASFV and evaluated its effectiveness and safety as a vaccine candidate in mice. A homologous recombination fragment containing ASFV CD2v was synthesized and co-transfected into HEK 293 T cells, a knockout vector targeting the PRV TK gene. The transfected cells were infected with PRV-ΔgE/ΔgI, and the recombinant strain (PRV-ΔgE/ΔgI/ΔTK-(CD2v)) was obtained by plaque purification in Vero cells. The expression of ASFV CD2v in the recombinant virus was confirmed by sequencing, Western blotting, and immunofluorescence analysis, and the genetic stability was tested in Vero cells over 20 passages. The virulence, immunogenicity and protective ability of the recombinant virus were further tested in a mouse model. The PRV-ΔgE/ΔgI/ΔTK-(CD2v) recombinant strain is stable in Vero cells, and the processing of CD2v does not depend on ASFV infection. The vaccination of PRV-ΔgE/ΔgI/ΔTK-(CD2v) causes neither pruritus, not a systemic infection and inflammation (with the high expression of interleukin-6 (IL6)). Besides, the virus vaccination can produce anti-CD2v specific antibody and activate a specific cellular immune response, and 100% protect mice from the challenge of the virulent strain (PRV-Fa). The detoxification occurs much earlier upon the recombinant virus vaccination and the amount of detoxification is much lower as well. The PRV-ΔgE/ΔgI/ΔTK-(CD2v) recombinant strain has strong immunogenicity, is safe and effective, and maybe a potential vaccine candidate for the prevention of ASF and Pseudorabies.
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