The recombinant pseudorabies virus expressing African swine fever virus CD2v protein is safe and effective in mice
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RESEARCH
The recombinant pseudorabies virus expressing African swine fever virus CD2v protein is safe and effective in mice Zhihua Feng1, Jianghua Chen1, Wangwang Liang1, Wenzhi Chen1, Zhaolong Li1,2*, Qi Chen1* and Shaoli Cai1*
Abstract Background: 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. Methods: 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. Results: 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. Conclusions: 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. Keywords: African swine fever virus, CD2v, Pseudorabies virus, vaccine, immunity Background African swine fever (ASF) is a highly contagious viral disease, a disease that must be reported to the World Organization for Animal Health (OIE) [1]. ASF was first reported in Kenya in 1921 [2], and then gradually *Correspondence: [email protected]; [email protected]; [email protected] 1 Fujian Key Laboratory of Innate Immune Biology, Biomedical Research Center of South China, Fujian Province, Fujian Normal University Qishan Campus, College Town, Fuzhou 350117, People’s Republic of China Full list of author information is available at the end of the article
spread to neighboring countries, in 2007–2017 to Georgia, Russia, Romania, and other European countries [3], and in 2018 to China, Mongolia, Vietnam, Cambodia, Laos, North Korea, and South Korea and other Asian countries [4, 5]. The disease
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